i, l eiiiadz - dtic · the number of osteoclasts and heightens their ltic activity. ccnbat injury...

I, L EIIIADZ0

Reports Control Symbol RCS-MEDDH-288 (R1)

ANNUAL PROGRESS REPORTFISCAL YEAR 19*9

( October 1*f - 30 September 9 )

DTIC-ELECTE

MAY 2 0 1987

APPROVED FOR PUBLIC RELEASE: DISTRIBUTION UNLIMITED

UNITED STATES ARMY INSTITUTE OF DENTAL RESEARCHWALTER REED ARMY MEDICAL CENTER WASHINGTON, D.C., 20307

7 5 ij,,

AD

UNITED STATES ARMY INSTITUTE OF DENTAL RESEARCHWALTER REED ARMY MEDICAL CENTER

WASHINGTON, DC 20307-5300

ANNUAL REPORT(I October 1981-30 September 1982)

Thomas P. Sweeney, COL, DC

October 1982

Supported by

US ARMY MEDICAL RESEARCH AND DEVELOPMENT COMMANDFort Detrick, Frederick, Maryland 21701-5012

DOD DISTRIBUTION STATEMENT

Approved for public release; distribution unlimited

The findings in this report are not to be construed

as an official Department of the Army position unless

so designated by other authorized documents

NTIS CRA&JDLYIC TAB[

J~jttlijt.) Gc'-

Dt

4L|

SECURITY CLASSIFICATION OF THIS PAGE (I#?.n Dat. Entered)

READ INSTRUCTIONSREPORT DOCUMENTATION PAGE BEFORE COMPLETIN4G FORMI. REPORT NUMBER __F 2 O VT AC CESSION NO0 3. RECIPIENT'$ CATALOG NUMBER

NEH-288-l APA~ / "a c4 _ _ _ _ _ _ _ _ _

4. TITLIE'lnd Subtitt) S. TYPE OF REPORT & PERIOD COVEREDAnnual Report

S. Army Institute of Dental, Research (1 October 1981-30 September 982).nnUal1 Progress! Report FY& V: S. PERFORMING ORG. REPORT NUMBER

7. AUTHOR(s) S. CONTRACT OR GRANT NUMBER(.)

aTIOaS P. SWeeney, COL, DC~

9. PERFORMING ORGANIZArION NAME AND ADDRESS 10. PROGRAM ELEMENT, PROJECT, TASK

Ab6WORK UNIT NUMBERSJ. S. Army Institute of ental Research 6 2BThS10 Task A;hingon, X 2007-50018162775A825 Task AA, AB;

______________________________________3M162734A875 Task AQIt. CONTROLLING OFFICE NAME AND ADDRESS 12. REPORT DATEL S. Army Medical Research & Development Ccumand October 1982

13. NUMBER OF PAGES

Fort etrick, Frederick, MD 21701-5012 614.MONITORING AGENCY NAME & AODRESS(Il different from Controlling Office) IS. SECURITY CLASS. (of this report)

UI'CUASSIFIED

15s. DECLASSIFICATION/OOWNGRAOINGSCHEDULE

IS. DSTRIBUTION STATEMENT (of Cha. Report)

pwroved for Public Release: Distribution Unlimited-

17. DISTRIBUTION STATEWMT (of th. abstract angiered In Block 20. If different from Report)

IS. SUPPLEMENTARY NOTES

19. KEY WORDS (Continue on rer.. side if necessery antd Identity by block number)hKou StEtts aeMtlAly;BoerdbeCrmc idgaalh~ r Bidgaalhuy~s atn cuay ells raeae

raIc lc;CrncEvcs ie;Cl ~te uvy c~sstou tratie;ti; aseMetl Bandae; iorrosioe ofralloy; Cideraioabillymr; icdegaabluid;oCyclicrs ; astn Acc ttiny Celltrosent; acetate

steobjetives arpler tooymin Bnoatndae byroio the Aecious; ofclnifcalan

ark; is dvied acrFd; toli the; ajrdental speialtiestruent;si isntlaediSeRAses aCndinjuri thatssare rec Ievify ittlakbe o ostd yciiin

DD ojec 1473 naITION OFeta Injr NOV 6515a DeSnLETEry.

SEURT CLSIIAINOjHSPG We e.Etrd

k D1

SECURITY CLASSIFICATION OF THIS PAGE("mn D84- Entered)

Item 19 continued:

Emergencies; Dental X-ray; Diphosphoinoeitide-Lysozyme; Gentamicin; GingivalExudates; Granular Tricalcium Phosphate; Herpetic Lesions; High VelocityMissile Effects; Impression Materials; Infection Control;. Inhibition of BoneResorption; Investmsnt Techniques; Laboratory Animals; Lidocaine; Lip Pathol-ogy; Lymphocytes; Marginal Leakage; Maxillofacial wounds; Microencapsulated

'Antibiotics; Monoclonal Antibodies; Nerve Agent; Oral Health Status; PolymerDevices; Porcelain-Metal Bond; Post and Core Restorations; Prostaglandin;Radionuclide X-ray System; Recurrent Aphthous Stomatitis; Reinforced Resin;Resin Restorative; Salivary Amylase; Salivary Enzyme; Salivary Physiology;Secondary Missiles; Segmental Mandibular Defects; Soldering Base Metals;Sporicidin; Storage Stability; Subtraction Radiography; Sustained ReleaseAntibiotic; Tracheal Grafts; Tricalcium Phosphate; Wound Healing; Xerocheili-osis.

Item 20 continued:

research groups, and the work is aimed at providing better preventive measureswell as treatment.

DA Project 3S162775A825 Combat Maxillofacial Injury.'Tasks AA, AB

-h e objectives are to develop simplified procedures for the care of complexillofacial wounds and injuries which require long time-consuming procedures

or reconstruction; to achieve minimal morbidity rates from oral emergencies,0reventable oral disease, and restorative failures; and to develop more effi-

ient, simplified, effective clinical and laboratory techniques which willsult in better utilization of manpower and a saving in time and materiel.

Project 3M162734A875 Medical Defense Against Chemical Agents.k AQ

<--' e objectives and purposes are the development of the basic scientific dataired for systems of soldier CW agent antidotes, soldier/patient decon-

amination, and medical management of CW casualties. ,.

ii

SECURITY CLASSIFICATION OF THIS PAGE(Whn DOta Entered)

FOREWORD

IN CONDUCTING THE RESEARCH DESCRIBED IN THIS REPORT, THE

INVESTIGATORS AICHERED TO THE "GUIDE FOR THE CARE AND USE OF

LABORAIORY ANIMALS" AS PREPARED BY THE COMMITTEE ON CARE AND

USE OF LABORATORY ANIMALS OF THE INSTITUTE OF LABORATORY

ANIMAL RESOURCES, NATIONAL RESEARCH COUNCIL.

4Li

USAIDR PROJECTS, TASKS, AND WORK UNITS

Page No.

3M161102BSI0 MANAGEMENT OF DENTAL INJURY AND CCMBATDENTISTRY.

A Dentistry

CA OB 6037 Acceleration of Wound Healing 1

DA OD 6021 Problems Involved in Military Oral Health 8Care Delivery Related to Therapeutic Agentsand Materials.

DA OF 6024 Identification and Control of Oral Infections 11(of Military Importance).

I OG 8676 The Secondary Effect of High Velocity 15Missiles on Craniofacial Tissues.

3S162775A825 CCMBAT MAXILLOFACIAL INJURY.

AA,AB,AC,AD Oral and Maxillofacial Sciences.

iA OE 6022 Preventive" Dentistry Measures of Military 18Significance.

DA OG 6033 Development and Evaluation of Dental 21Materials and Materiel for Army Use.

IA OG 6034 Development and Improvemnt of Metallic 26Restorative Materials.

DA OG 6679 Initial Treatment of Ccvnbat Wounds 30

DA OG 8670 Development and Evaluation of Dental 33Materiel for Field Use.

- -. .. ... . . .. . ...... .. - 4L

USAIDR PROJECTS, TASKS, AND WORK UNITS

Page No.

DA OG 8672 Epidemiological Investigation of Dental 37ENergencies.

[A OG 8674 Development and Evaluation of Methods and 39Materials for Treating Traumatic Injury.

[A OH 6030 Natural History of Oral Lesions Encountered 41in the Soldier.

[A OH 6037 New and Inproved Techniques For Grafts and 43Bone Regeneration in Traumatic Wounds.

[A OK 6020 Biodegradable Materials For the Treatment of 46Fractures and Soft Tissue Wounds in theMilitary Situation.

3462734A875 MEDICAL DEFENSE AGAINST CHEMICAL AGENTS.

AQ Pathophysiology of Chemical Agent Poisoning.

DA OG 0717 Study of Saliva as a Diagnostic Tool for 49Presence-6f Lethal Agents.

USAIDR PUBLICATIONS 51

DISTRIBUTION LIST 55

| AQENCV ACCESIOII 1S DAVE Of SUS ANMAS aspoor COmRmOL IVAOL

RESEARCH AND TECHNOLOGY WORK UNIT SUMARY DA OB 6037 82 10 01 DD-DR&E(AR)S36

L. GAVE PREtv sMN 4. 111M0 of SUMMARY S. SUMMARYICV SCUNEY 7 mReRaene ta DownV IMsToR E. SPCIC DATA-. LEVEL of SUM

81 100 D. CHANGE CYCNL T cSS N

IS. NO.aCOOES.* PROGAM ELEMENT PROJECT NUM6ER TASK ARMA MUNImER WORN UNIT NUMBER

.6 P*N I 61102 3M61102BSI0 D6116 CONT089TIN*

AA~~o 111117M7 015lG 80 7.2:511. TITLE (P*.4E M E I5 .- B c~d.t.

(U) Acceration of Wod HealingIL SCIENTIFIC ANO TCCNM*LOO4CAL AREAS

002300 BiochemistryI. START DATE 14. E8TIMATIO COMPLITO OATE I& FUMOIMN AGENCY I. PERFOMANCE METNO

66 07 CONr DA C. IN-HUSEITC CONTRACT/ORANT M. RESOUtCS ESTIMATE 5 PROFESSIONAL MAN YR k FUNDS (IN V_ ld)

IL OATIGFECTIvE 8 ESPlRATOOU

b. NumE,* FISCAL 82 2.0 174a TYP44 a AMOUNT' YEAn wwwo[N

& SISOF AWARI t. CUDA. AMT. 83 2.5 195SE. RELSPONUPNLEK 000 ORGANIZATION I. PEIRIN I~6RZT

"- USA Institute of Dental Research MAR, USA Institute of Dental Research

A *.. Washington, DC 20012 AooRI",* Washington, DC 20012

PRINCIPAL INVESTIGATOR (FWIM SAR 1 U.S. A~d-1 f-.II.MN

R-,SPONMM INOVIDUAL A,,* Hollinger, J., LTC, DCNAME.8eeney, T.P., CODL, DC ,EEPoN g: 202-576-3764TELE N01, 202-576-3484 SOCIAL SECURITY ACCOUNT ,UM6R R)C :DA

I.M tEERS USJ:'OITIIVEE S. Miller R. BS

Foreign Intelligence Considered . Miller, R., B.S..MAWvnkIC. J. CPTDCX I9IEORDS (P-.--. ACN .,* $~Or coi .... c.1 (U)Diphosphoinositiae-Lysozyme tU)uitric Acid k0)Prossa-

glandin (U)Copolymer (U)Inhibition of Bone Resorption (U)Cyclic AMP (U)Laboratory Animalft TEC*ICAS. OSJCTIVE.* I& APPROACN. SIL PROSRERI (F-waf JNIUSW o-dmip MANNR. or - PRMd .1 .1 ." RIM ftewfr CI..IN*M c.".)

23. (U) Recent studies show that 10-12% of combat wunds involved the maxillofacialapparatus. Further, 7% of noincombat injuries requiring hospital care involve the max-illofacial region. This results in the loss of approximately 1,000,000 man-hours peryear. The research objective is to accelerate or otherwise improve the healing ofmaxillofacial injuries.

24. (U) Studies on the effects of biochemical and physical factors to include chelatecomplexes, cyclic AMP, prostaglandins, and in vivo growth factors on the rate of heal-ing in soft tissue and bone will be done. The mechanism of any beneficial alterationin healing effected will be invested and pursued to human usage.

25. (U) (81 10 - 82 10) A diphosphoinositide-lysozme (DPIL) complex was found toaccelerate the healing of osseous wunds in rats as did a combination of polylactic(PLA) and polyglycholic (PGA) acid (50:50). Ccmbined.DPIL and 50:50 PLA-IGA was alsoevaluated in the same system. Available data indicate a greater acceleration of osse-ous healing with combined DPIL-copolymer than with either alone. Data collection iscontinuing. Combined PIA-PGA with Dilantin did not accelerate osseous healing. Studieson the effects of PGE2 and cyclic nucleotides on osseous and pulpal healing are alsoin progress. Citric acid was found of value in accelerating the healing of gingivaltissues. Eforts continue to isolate a powerful inhibitor of bone resorption. A 50%inhibition Ca release from fetal long bones has been obtained. The nature and modeof action of this inhibitor remain to be determined.

Fos f GUAO PRVOUS EDITIONS2 OF THIS FORM ARE OBUCLEKTE. 00 FORMS 1SSNA NO pov sDI MARAI ANO 145.1. I MAR E IP OR ARY U1I ARE O8SOLfTE

L

PROJECT: 3MI61102BS10

WORK UNIT TITLE, ACCELERATION OF WOUND HEALiNG

PRINC IPALINVESTIGATOR: LTC JEFFREY 0. HOLLINGER, DC

A Study on the Behavior of Bone in Organ Culture With

Special Emuphasis on PGE 2 and Indomethacin

PROBLEM4: There is abundant clinical and experimental evidence that bonecan synthesize PGE , and that local synthesis of this prostaglandin leadsto local bone resoiption. It is believed that PGE causes an increase inthe number of osteoclasts and heightens their ltic activity. Ccnbatinjury can produce mechanical stress to periosteum, causing perturbation ofthe tenant cell membranes with a resultant increase in the synthesis andrelease of prostaglandins of the E series. Surgical treatment and healingof osseous wounds would have a more predictable and reliable incidence ofrepair if the endogenous prostaglandin E series of lytic agents could bethwarted. Indamethacin has generated considerable attention due to itsremarkable ability to inhibit PGE and PGE 2 synthesis. If indcmethacincould effectively inhibit PGE Synkhesis by2 bone in vitro, this would bethe first step to better understand PGE 2 and a potential in vivo inhibitor.

APPROACH: A total of 192 calvaria from D/O neonatal CD-I mice were asep-tically removed and placed in plastic media dishes - one calvaria per dish.The dishes were gassed with a mixture of 5% CO and air. The calvaria weredivided into four treatment groups: (1) Control: no exogenous agents;(2) Indomethacin: 200 ng/ml of medium; (3) PGE : 100 ng/ml of medium; and(4) Combination: 100 ng/ml PGE plus 200 ng/ml i~dcmethacin. Calvaria wereharvested at 24, 48, and 72 h irs and were processed for histamorphametricevaluation, radioimmune assays of PGE2 and cyclic C1MP and AMP, and foratomic absorption spectrophotcmetry for calcium and phosphate molar ratios.

RESULTS AND DISCUSSION: Assays have not yet been finished. The histcmor-phometry analyses have been completed and do not support the findings ofHoltrop, Raisz, and King - that PGE stimulates pre-existing osteoclasticactivity. Rather, PGE increased i3 le number of osteoclasts - a findingsimilar to Schelling, Wolfe, and Tashjon. Also, indomethacin added tocontrol dishes inhibited spontaneous basal bone resorption. Indamethacinhad no effect on calvaria osteoclasts in dishes when exogenous PGE wasadded to the medium. Further studies are under way to evaluate PGE siriesinhibitors; these include: ficose-5,8,11,14 tetraynoic acid, 5-oxa and7-oxa-pyrostanoate derivatives, linolenic acid, phenylbutazone, anddichloramethane diphosphonate.

2

PROJECT: 3M161102BSI0

WORK UNIT TITLE: ACCELERATION OF WOUND HEALING

PRINCIPALINVESTIGATOR: LTC JEFFREY 0. HOLLINGER, DC

Enhanced Healing of Soft Tissue Wounds Using Diphenyihydantoin

Sodium Incorporated Into a Meshwork of Biodegradable

Copolymer (50:50 Poly L-(-)Lactide-co-Glycolide)

PROBLEM: High velocity projectiles frcn military weapons produce a widevariety of tissue destruction. Frequently, significant amounts of softtissue are avulsed. These sites often require extensive skin grafting.Unfortunately, bulk replacement material for the connective tissue bed isscarce. The resultant healing site is "cupped-out," concave, and grosslyunesthetic. A material that is bioccipatible and that can induce collagenformation to "bulk-up" the connective tissue bed will insure for anesthetic effect. In addition, if the material can hasten collagen forma-tion in the wound site, this may prove to be an important aid in facili-tating the healing of surgical and traumatic wounds.

APP ACH: The copolymer of 50:50 poly L-(-) lactide-co-glycolide wassolubilized in methylene chloride and diphenylhydantoin was added, andusing nitrogen, a fibrous mesh dressing was prepared. The dressing wasplaced onto surgical wounds created on the backs of rats. Healing wascapared to plain copolymer, untreated, and IP-diphenylhydantoin-sodiumtreated wounds. Wounds-were evaluated by assaying for total proteinhydroxyproline; using RIA for Dilantin released in the animal; by tensiletesting; and by histcmorphmetry.

RESULTS AND DISCUSSION: After 28 days there was no significant differencein the rate of healing between any of the four treatment groups. However,animals that were receiving the IP-diphenylhydantoin-sodium did display agreater amount of collagen and a higher density of fibroblasts than theother three treatment groups. It was also apparent that the physicalproperties of the copolymer dressing were not as favorable as had beenanticipated. By 3 days, most dressing material had beccime brittle and hadbegun to fray. Therefore, it could not be closely apposed to the healingconnective tissue bed. Consequently, the release of diphenylhydantoin-sodium into the healing sites was inadequate. A supple, cellophane-typecopolymer dressing was prepared and evaluated to militate against such aproblem.

3

PROJECT: 3M161102BS10


PRINCIPALINVESTIGATOR: LIU JEFFREY 0. HOLLINGER, DC

A Study on 50:50 PLA:PGA+DPI-L for the Prcmotion of Calcification

in Osseous Defects

PROBF24: Approximately 10% of ccmbat injuries are in the maxillo-mandibu-lofacial region. For a casualty of this type, establishment of structuraland anatomical continuity is an extremely important goal in the earlymanagement and restoration of function. Current treatment of avulsive andfracture wounds of the maxillo-mandibulofacial region includes a variety ofimplant materials (autografts, allografts, xenografts) and an array ofmetallic fixation devices. Sone materials may initiate an adverse immuneresponse. Most metallic fixation devices eventually require surgicalremoval. Both situations are unacceptable. If a biocompatible, biodegrad-able implant could be developed, then adverse host reactions would notoccur and surgical removal would not be required.

APPRlACH: In an attempt to develop a bioccrpatible, biodegradable implantaterial that could initiate osseous healing, copolymers of polylactic andpolyglycolic acidL in combination with an acidic phospholipid-lysozymecomplex were evaluated. Each component of the implant material wasexamined for host acceptability and bone inductive effects. Endochondraland intramembranous bone defects were treated in experimental animals. Inseparate experiment the - acidic phospholipid-lysozyme ccmplex and thePLA:PGA (in a 50:50 ratio) were individually assessed at the host sites inthe following ways: histcmorphcmetrically, histochemically, by enzymeanalyses, by total protein and hydroxyproline ass Is, and by elementalanalyses. After the data analyses for these experiments, the combinationof copolymer (50:50 PIA:PGA) and acidic phospholipid-lysozyme was evaluatedin a similar fashion.

RESULTS: In experimental animals, the acidic phospholipid-lysozyme coplexinduced healing of osseous wounds at a more accelerated tempo than woundsreceiving no treatment. In a separate experiment, at 7, 14, 21, and 28days the copolymer treated bony wounds had healed more rapidly thanuntreated wounds. By 42 days, the rate of healing was similar between thetreated and untreated groups. All phases of the evaluation of the combi-nation type implant have not yet been ccmpleted. The currently availabledata indicate that the copolymer-acidic phospholipid-lysozyme ccmplex doesinduce calcification at a more rapid rate than the untreated or singlereagent treated osseous wound.

4L4

PROJECT: 3M161102BSI0


PRINCIPALINVESTIGATOR: LTC JEFFREY 0. HOLLINGER, DC

A Study of Prostaglandins and Cyclic Nucleotides

and Their Reaction to Pulpal and Periapical Pain

PROBLEM: Prostaglandins are one of the most prevalent of the autacoids.Their synthesis is orchestrated by astonishingly diverse stimuli. The

prostaglandins produce a remarkably diverse spectrum of effects, one ofwhich is as a mediator for pain. Pain is one of the most singularly

insidious modalities responsible for eroding both cognitive and actively

dexterous skills. Cephalic pain, and specifically toothache pain, areprofound stress inducers that create havoc with the soldier's psyche. By

isolating and identifying pain mediating biochemicals endogenous to patho-logical pulpal and periapical tissues, it may be possible to militateagainst their synthesis, thereby obliterating certain types of dental pain.Elimination or modification of pain will help to insure for a soldier'shigh performance in activities that require recognition and dexterity.

APPROACH: Tissue samples will be retrieved from four sources, with at

least fifteen specimens from each source. There will be four categoriesof sources: (1) Vital asymptomatic pulp; (2) Vital symptomatic pulp;(3) Asymptcmatic periapical lesions; and (4) Symptcrnatic periapicallesions. After removal, all tissues will be immersed immediately in LN ordry ice and will be stored at -40C until needed. Assays will consis ofRIA's for PGE1 , PGE 2, PGF2d' cAMP, and c" P.

RESULTS: The study is at the tissue collection stage. Assays have not yet

begun.



PRINCIPALINVESTIGA TOR: COL STEPHEN G. WOODYARD, DC

Evaluation of Citric Acid Enhancement of Oral Soft Tissue

Healing on Previously Denuded Tooth Root Surfaces

PROBLEM: Early return of the injured soldier to duty is of paramountimportance, placing an emphasis upon methods and techniques which canaccelerate wound healing in the maxillofacial area. Contamination of toothsurfaces, soft tissues, and bone subsequent to wounding in the oral regionoccurs routinely due to the presence of saliva and oral microorganism inthat area, complicating the healing process. Of particular importance areroot surfaces of teeth partially avulsed or exposed by loss of adjacentsoft tissues and bone, contamination resulting in delay and prevention ofhealing of soft tissues and bone placed adjacent to these surfaces duringsubsequent surgical repair. Mechanical debridement of contaminated toothsurfaces prior to coverage with soft tissues and bone is relativelyineffective, often resulting in subsequent loss of such teeth. Studiesccmparing mechanical debridement to a combination of such treatment andlavage with citric acid indicated acceleration of wound healing occurs inuncontaminated oral wounds involving tooth surfaces, but little data existsrelative to its usefulness in contaminated wounds.

APPROACH: Six cynamolgus monkeys had facial root surfaces surgicallyexposed by removal and repositioning of soft tissues and bone. Six weeksof exposure and contamination of tooth surfaces by saliva, oral micro-organisms, and their products followed prior to surgical repair. Toothsurfaces were treated by either mechanical debridement alone or in combi-nation with topically applied pH 1.0 citric acid for 3 minutes. Softtissues were placed over treated root surfaces and animals were sacrificedat varied time intervals up to 42 days postsurgical repair. Areas includ-ing treated teeth, soft tissues, and bone were removed, fixed, and preparedfor histologic and bicmetric evaluation.

RESULTS: All areas treated with both mechanical debridement and citricacid healed rapidly with new connective tissue attachment, much as existedprior to creation of the wounds. Conversely, mechanical debridement aloneresulted in incomplete repair with an epithelial interface between softtissues and tooth surfaces. No adverse soft tissue or bone responsesoccurred subsequent to citric acid application. Study indicates use ofcitric acid following conventional mechanical debridement of contaminatedroot surfaces is advisable, resulting in both acceleration of soft tissuehealing and improvement of the final healing result.

6

PROJECT: 3MI61102BS10

WORK UNIT TITLE: ACCELERATION OF WOUND HEALIMG

PRINCIPALINVESTIGATOR: JAMES R. HEATH, III

Identification of Leukocyte Populations Responsible for Production

of OAF and Their Role in Bone Resorption

PRO)BLE4: There is a retardation of osseous repair in wounds that becomeinfected, a common occurrence in traumatic combat injuries - especiallythose of the maxillofacial apparatus. One of the factors thought to beresponsible for this delayed healing is the lymphokine osteoclast activat-ing factor (OAF). OAF is produced by lymphocytes in response to stimula-tion by by bacterial products and results in activation of local osteo-clasts which in turn leads to enhanced bone resorption and retardation ofosseous repair. OAF has also been implicated in the osseous deteriorationseen in periodontal disease and in multiple myelcma patients. An investi-gation into the mechanism of production and modes of action could lead toabrogation of the detrimental effect of OAF and thus a quicker return ofthe soldier to duty.

APPROACH: OAF is produced by stimulating human lymphocytes with theitogen phytohemagglutinin in large volume cell culture. The nanogramquantities of OAF in the cell culture supernatants are concentrated andpartially purified by an ultrafiltration protocol developed in our labora-tory. Further purification is accomplished by a variety techniquesincluding ammonium sulfate precipitation, gel filtration, ion exchangechromatography, high performance liqid chrcmatography, and polyacrylamideelectrophoresis. Osteolytic activity (OAF) is detected in culture super-natants an 5 purification fractions by a bioassay system involving therelease of Ca from fetal rat long bones in organ culture.

RESULTS: Osteolytic activity has been found in cell culture supernatantsand in gel filtration fractions representing a variety of molecular weightsfrom 18,000 to approximately 1,500 as estimated by gel filtration. Theseresults agree with those of others suggesting that OAF exists in a systemof freely associating-disassociating active sub-units. The amount ofdisassociation increases with increasing ionic strength of the medium. Anunexpected discovery was the presence of an apparent powerful osteolyticinhibitor in the lymphocytes culture supernatants, reported in FY 80-81.The presence of this inhibitor has been cenfirmed in all subsequent culturesupernatants and its molecular weight is estimated to be slightly less than6,000. This inhibitor hq been further purified and results in greaterthan 50% inhibition of 4 Ca release from fetal rat long bones in thebioassay system. More work is needed to determine the nature and mode ofaction of this inhibitor and its possible role in vivo.

4L7

I. AseNCT ACCESIION Ia. DAVe of SJINA.'Y aoar c~mrog svumoLRESEARCH AND TECHNOLOGY WORK UNIT SUMARY DA I) 6021 82 10 01 DO.DR&E(AR)J6

A I 4.41O OF 1A1W SUNARV SCYr wo, AAmNS -'I. MC.oN.m4e

•N pg4 , c o AT.. I.L . SUNT-7_IS___________________TRACTOG ACCESS

811001 D. CHANGE U NL 0 LaIa. N@./Coots.$ PROGRAM ELEMENT PROJECT HUNGER TASK AREA MWUER WORK UNIT Mumma"

"6 PRI"AR 61102A -3=1L1 D2o oA HS,'b. C~mvumuviuo----

I IIIIIIIII IOG 80-7.2:5"TITLE " - wo S-Mmev cf "- C.*' (U) Problems Involved in Military Oral Health Care DeliveryRelated to Therapeutic Agents and Materials

IL SCINTIFIC ANO TECHNOLOGICAL AREAS

012600 Pharmcology 002300 Biochemistry 010100 MicrobiologyIL SITA? i DAE1 STIMATED COUPLETION DATE IL FUNDINO A9ENCY IN REFORMAKCg METHOO

68 IN F CONT DA IC. IN-IUCII0TNACT/ONANT 1. RESOUNCE$ ESTIMATE POFESSIONAL 1AN VRS h FUNDS I. 0-41.jt)

IL ES/EPPECTIV9, CXPINAIC.:

16 nufteca' VISCAL 82 2.0 160& TYP: & AMOUNT: YEA .

&.-,0SF .AR t.CUM. AMT. 83 2.5 175.... PONML, DOD 04ANI.ATION J. P90 1.S ..... o .. ATION I

*a" LA Institute of Dental Research N"': USA Institute of Dental Research

A R." Washington, DC 20012 ADooSI* Washington, DC 20012

PFINCIPAL INVESTIGATON (P-WA UAN it U.S. A"l4 f..II..S.

OsPO ,OLE #NDIVIOUAL --a:- Setterstram, J., Ph. D.NAE Sveeey, T.P., OL, DC TLEP"ONE: 202-576-3662TELP6ONE: (202) 576-3484 SOCIAL SCUNIT. ACCOUNT -.. ,0: POC:DA

I1. GENERAL USE ASSOCIATE INVESTIOATORS

Foreign Intelligence Considered A.,: Vincent, J., LIU, DC Miller, R., B.S.8. K.oo1.NAE: P J., CPT. DC Heath, J., M.S.

KTUONOS( .A , , ......... U) Microencapsulated Antibiotics; (U) Infection Control ;(U) Wound Healing; (U) Sustained Release Antibiotic; (U) Laboratory Animals28. TIICHNCAL OSJECTIVE.N 21. IIPACII. S4. POOGNESA (P4Wf I-a-fw... ~~a . JIHd 6, -.I. P ... I ma .. -I I...iE, CfI.NYff. C&I.

23. (U) To evaluate the specidl military problems of drug storage, heat susceptibility,long-term drug potency, sterility of bulk items, lack of refrigeration in combat zonesand delivery to the patient. To investigate drug hazards. To investigate the use ofbiodegradable polymers for the long-term, slow release of drugs.

24. (U) Improved means of drug delivery in the field using microencapsulated medicamentswill be studied. Mle hazards in the use of various drugs and the use of biodegradable,bioccmpatible materials for surgical repair of combat wounds will be studied.

25. (U) (81 10 - 82 10) Although as previously reported microcapsules made of 68:32DL-PLGA effectively eliminated wound infections in rats, they biodegraded too slowlyand released drug for too long a time. Most recently, microcapsules have been polymer-ized from a 50:50 DL-PLGA excipient using a better polymerizatin catalyst and purermonomers. These microcapsules met the target duration of release of 14 days and arepresently undergoing in vivo evaluation. Geptamicin sulfate microcapsules preparedfrom 68:32 DL-PLGA dumped drugs too rapidly and failed to eliminate infection in theinfectious rodent model. A more extensive development program will be required tosolve problem inherent in encapsulating this drug.

D D ,. . I4 98 PRVOUS DITIONS OF T.IS FORM ARE Ol CLETE 3 50 PORM S 1446A I NOv S

IMM ANO 14001 I f MA1 95 IIO AN94YUSI iARC O61L E rT

8

PIOJECT: 3M161102BS10

WORK UNIT TITLE: THE PROBLEMS INVOLVED IN MILITARY ORAL HEALTH CAREDELIVERY RELATED TO THERAPEUTIC AGENTS AND MATERIALS

PRINCIPALINVESTIGATOR: JEAN A. SETTERSTROM, PHD

Biological Activity Verification of Specified

Microencapsulated Antibiotics In Vivo

PROBLEMS: Combat wounds are characterized by a high incidence of infectionprimarily because of the inevitable presence of devitalized tissue andforeign body contaminants from missile fragments that carry dirt and debrisinto the wound. During evacuation, the wound may be exposed to furthercontamination and delay before initial treatment. Wound healing in thecombat casualty, therefore, must overcome adversities not seen in thehighway victim or civilian counterpart. Among soldiers, infections haveremained a major cause of morbidity that results in lengthened hospitali-zation and cambat ineffectiveness.

APPROACH: Improved methods to deliver antibiotics to contaminated tissuefollowing traumatic injury are needed in order that sustained and effectivetissue levels of antibiotics can be maintained at the wound site in spiteof the inadequate perfusion of blood resulting fran shock or the destruc-tion of blood vessels to devitalized areas. The improved method should beeasily applied in a single dose to the wound site as soon as possible afterinjury when infection is most likely to be suppressed. Such a novel anti-biotic delivery system is being developed in which ampicillin anhydrate andgentamicin sulfate are being incorporated individually into microspheres ofbiocampatible, biodegradable, copolymer that are formulated to slowlyrelease the drug over a sustained period (14 days). These microsphereswhich will completely biodegrade once all drug is released exist as a free-flowing powder that can be easily dusted onto wounds under field condi-tions.

RESULTS: Studies to formulate microcapsules with optimal drug releasekinetics have been ongoing. Prior to this year, microcapsules were pre-pared fram a 68:32 DL-PLA:PGA excipient that biodegraded about 30-50 dayspost-treatment giving an undesirably long release of drug. Most recently,microcapsules have been polymerized fram a 50:50 DL-PLA:PGA excipient usinga better polymerization catalyst and purer monamers. These microcapsulesmeet the target duration of release of 14 days. By carefully standardizingmaterials, methods and processes used in the design of these microcapsules,and following successful in vivo efficacy evaluations, clinical testing ofthe ampicillin microcapsu1-sFi7uld be possible by 1984.

The success achieved with encapsulated ampicillin in controlling woundinfections in a rodent model has stimulated interest in the encapsulationof other antibiotics. Encapsulation of additional antibiotics are desir-

9

able so that encapsulated mixtures can be formulated to assure broad spec-trum antibiotic management for wound therapy. During this year, gentamicinsulfate microcapsules prepared with a 68:32 DL-PLA excipient were produced,however, results of in vitro and in vivo evaluations indicated that thesemicrocapsules dumped drug too rapidly to be effective in controlling woundinfections. A more extensive development program will be required to solveproblems inherent in the encapsulation of this drug.

4LA

0. a06.CW *441m &save @# UUU*4t 7*~ a"& vu

RESEARCH AND TECHNOLOGY WORK UNIT SUMMARY DA F 02 8 1 0 D.M&(ARAt*

811001 D. CHANGE U ~ ...... UI R I.00!I06 @.C@040a 000AM CLU10496 PUeJeCT "W"460 TAGM ANCA WU660 wo" eN WI.? ft

I61102A Ifl61l02R51f6

a STOG 90-7.2:SI L TOWLS 4D.* -00 8-*Claa04g C-"b3

(Ui) Identification and Control nf Oral Inferrian.IL SCIUSTMfC "S a C36OOESCA4 a*Aa*

0,10100 Microbioloizy; 00230 iceity ______________________

66 0 CON DAC. IN-HOUSEMI C0OtSACT16QnT W *fC* *ICLT~nT4 0.06100ma04 WV0S FOG 0uU. ~

U~~6"NTO eag.9UY.t %%-.am

UTVP. LaminuT, VCaM ufI.

ftAMGLOPA-a.. I. CUU. AMT. 832.0 154

a~*USA Institute of Dental Research USA Institute of Dental Research

Wahntn C2~2 Washington, DC 20012

MasUaet** Setterstrom, J.a- SWEENEY, T.P. Takavown, 202-576-3662TCLgPg.g. 202-576-3484 Moaat 84CuM00T AW"SV MMS44 POC :DA

.. Woodyard, S.mm.Vincent, J., Heath. J.

~ (U).Oral Health Status; (U) Gingival Exudates;(U) Crevicular Fluid; (U) Spor cidin- (U) rudy (ii) mjnnrs,1,.a ApftibodjoaA& tac.eCoa. 00b..VC~.e I& avacs. BIU P60M fqpw m.. " !~ aa e t . ~_~ - *0_ a .. C-0

23. (U) To investigate the source and* treatment of oro-facial infections encounteredin field conditions, foreign countries and diverse climates. To evaluate the specialagents, instruments and chemicals necessary under military conditiona.

14. (U) Oro-facial infections-of s ignificance in the diverse military environment will)e studied by microbiological, immunological, and electronmicroscopy methods. Possible;ources of oral infections will, be evaluated and the effectiveness of commerciallyvailable as well as in-house designs will be studied for their ability to control or:revent oral infections.

Z5. (U) (81 10 - 82 10) Development of a rapid non-immune biochemical technique forvaluating oral health status based on the hydroxyproline content or gingival exudates'as continued. Improved clinical collection methods and quantitative evaluation)rocedures have been developed and a baseline study on clinical variations is in)rogress. A clinical study of the relative efficacies of the cold sterilizing agents'poricidin and cidex is continuing. Thus far aporocidin has exhibited the more usefulroperties. A study has been initiated on the use of monoclonal antibodies for theapid identification of anaerobic microorganisms associated with maxillofacial woundnfections.

0 . .. 14 9~ 8t g.

PRaJECT: 3MI61102BS10

WORK UNIT TITLE: IDENTIFICATION AND CONTROL OF ORAL INFECTIONS

PRINCIPALINVESTIGATOR: COL STEPHEN G. WOODYARD, DC

Measurement of Collagenolytic Activity

in Areas of Chronic Infection

PROBLEM: A reliable, noninvasive means for evaluating tissue healing ordegradation is needed. Current state of the art methods are not signifi-cantly better than those used during ancient times which describe tissuecolor, swelling, temperature, and presence or absence of hemorrhage.Invasive methods such as biopsy are of same value in certain pathologicstates, but are of little use in measuring healing activity, this being aphenomenon essentially unmeasurable by visual observation of tissuesections as well as being contraindicated in healing wounds. Hydroxy-proline is a product of collagen degradation, found in no other tissuesother than a minor caponent of a serum camplement fraction, suggestingvariation of hydroxyproline levels would occur directly with inflammatorycollagenolysis. If such levels could be correlated reliably with knownlevels of tissue destruction and new or established clinical indices, morereliable evaluations could be made relative to tissue healing status andthe relative effectiveness of therapeutic regimens used in treatment of thewounded soldier in the field environment.

APPROACH: An area readily available for studies of chronic or acutedestruction in intraoral areas is the soft tissue/tooth interface termedthe gingival sulcus. Treatment of that highly contaminated area subsequentto acute infection provides an ideal model for studies on wound healing inintraoral wounds.

RESULTS: Data have been obtained that indicate that hydroxyproline levelsvary directly with increased areas of clinically measured destruction. Asignificant step forward was taken by the development of a reliable andhighly accurate method for processing collected exudates and evaluatingquantitative hydroxyproline levels. Improved clinical collection methodsto include improved quantitative measurements of exudate levels has beendeveloped which will allow accurate evaluation of concentration of hydroxy-proline in tissue fluids. A study is in process to determine base linelevels of hydroxyproline and diurnal variation in varying states of inflam-mation and tissue destruction prior to evaluating effects of various thera-peutic procedures.

12


WORK UNIT TITLE: IDENTIFICATION AND CONTRML OF ORAL INFECTIONS

PRINCIPALINVESTIGATOR: M. A. DEREVJANIK

Evaluation of Cold Sterilants

PROBLEM : (1) To determine which glutaraldehyde-based disinfectant,Sporicidin or Cidex, is superior in efficacy and cost effectiveness,(2) to determine the dilution and duration of effectiveness for each coldsterilant during clinical use, and (3) to determine the effect of storageconditions on the stability of the constituent chemicals of each agent whensubjected to temperatures equivalent to climatic extremes.

APPROACH: The cold sterilants were evaluated using the official methods ofanalysis of the Association of Official Analytical Chemists (AOAC) by per-forming the AOAC spore, bactericidal, and tuberculocidal tests. Solutionswere tested at different dlutions under various conditions. These condi-tions included timed sampling following shelf activation in the laboratory,as well as samples removed from disinfectant trays following use in theclinic. Phase 3 of the problem will be addressed during the next year. Inthis phase, samples of each cold sterilant will be subjected for known timeintervals to various humidities and temperatures to stimulate climatic con-ditions likely to be incurred under field conditions. Subsequently, thecold sterilants will be tested for efficacy.

RESULTS: Part 1 of the problem has been campleted and reported on pre-vi3 y9. Briefly, Sporicidin was found to retain an effective sporicidalactivity at dilutions five times, and an effective disinfection dilutionfifteen times that of Cidex. Evaluation of samples removed from disinfec-tion trays following use for 1, 2 and 3 weeks in the dental clinic showedSporicidin to retain sporicidal activity after 3 weeks of use. Cidex,however, failed to exhibit sporicidal activity after 2 weeks. Sporicidin,when diluted 1:5, retained the same sporicidal activity of full-strengthCidex. Bactericidal testing of clinically used disinfectant to determinethe lowest concentration of each agent that is bactericidal for a 2-3 weekperiod is ongoing.

13

PRO)JECt: 34161102BS10

WORK UNIT TITLE: IDENTIFICATION AND CONTROL OF ORAL INFECTIONS

PRINCIPALINVESTIGATOR: LTC JACK W. VINCENT, DC

Production and Utilization of Monoclonal Antibodies

in the Rapid Identification of Anaerobic Microorganisms

Associated with Maxillofacial Infections

PROBLEM: There is currently no rapid method for the identificationmicroorganism present in a wound exudate. Conventional techniques requireapproximately 2 to 3 days for this procedure. Such a delay in evaluatingcontaminated maxillofacial wounds and in selecting appropriate antibiotictherapy could result in severe sequelae. A method of rapid detection wouldappear to be ideal for the prevention of such a delay and may serve as amodel for the detection of biological agents which might be utilized in abiological warfare environment.

APPROACH: Mice will be immunized with selected microorganism which arecommonly associated with maxillofacial wound infections. The spleen willbe harvested and cells dispersed. These lymphoid cells will be used withmouse myeloma cells in polyethylene glycol to form hybridoma cells.Efforts will be directed to isolate a clone of cells possessing immortalityand secreting a monoclonal antibody specific for the microorganism ofinterest. When isolated, .this monoclonal antibody will be used to developa system for rapid deteZtion (2-3 hours) of wound contaminants. Such asystem (most probably the ELISA) must display extreme sensitivity andspecificity which should be provided by these techniques.

RESULTS: The organisms Fusobacterium nucleatum (ATICC 20953) andBacteroides gingivalis (ATCC 33277) have been selected and compared withclinical isolates of these organism by polyacrylamide gel electrophoresis.They were found to be identical. Separate mice were immunized with F.nucleatum and B. gingivalis and fusions are being performed with mousemyeloma cells.

14

I. 6CNCV 4CCd(sN6 18 @.I Of S0MMo A4 Aspoo coa wrwoL orm

RESEARCH AND TECHNOLOGY WORK UNIT SUMMARY DA W 8676 82 10 01 DD-DR&E(AR)6J6

L TC P*V SW V J KIO Of , 1aE L $£ AIu SCTm T CU Ty .fteE AUSI Iy wo IT 16k SPECIFIC OATA " . L'VCL OF SM81 10O K. CCM r ""U Wo IMS

te. MO./COcesI PROGNAM SLIEMIENT PfOJCCT N4UM9INR TASK ANEA NISCA WORK UNIT NUMQIR

0 ., 0R HM61OHIUA 3(54

• / ,1 I111/1111T -= 80-7.2:5t TL (P&-o ".-to as*4U~a- CdI1 (U) The Secondary Effect of High Velocity Missiles onCranofacial Tissues

IL SCIENTIFIC ANO TCCNNOL@OICAL ANIAS$

IL IVAT IPhy iolo IITI 0TI51 CMPITI1 OAT IL. FUNO"O AGENCY I4 PEVO ANCE TNO0

81 10 82 10 DA I C. IN-HOSE1T. CQ1TRACT/OWAAT %6. mgSOUNCES CSTIMATs PN OCSSIONA .. " VfS k 0 U" O. (I

a OAT C11 Iyve, 9ANATOON: '""' 1.4N

1L o."6.," ""SCA 82 0. 5 30

_______ ___o,_ (._CUM._T. 83 0 00s. ,M*g ,P*W I E 00 Omea NO R 1OO . P I.E OOWnNo O GANaI AT IO I

"a e USA Institute of Dental Research ""a" USA Institute of Dental Research

. ,. Washington, DC 20012 Af...,. Washington, DC 20012

OM14CIPAL MNVESTIGA TO* (F. &UA0 I#~ US. 44 .A i.I.

,IO,.E I,"SOIWMAL MAC:* Carpenter, W.M., cOL, DC.M Sweeney, T.P., COL, DC T,LCPOK 202-576-3080..L.. ., 202-576-3484 SCIAL ,,9.,, ACCOUNT NOSICN! FOC:DAI1 SHENERAL uSE ASSOCIATE InvEeWArTONs

Foreipn Intelligence Considered %AMC! Lewis, D., LTC, DCMANE:

23. ItEV11,,- A- ,-- .. ,.ISS...,or Cd.)(U) Cranofacial Tissues; (U) High velocity Misslie Effects

(U) Maxillofacial Wounds; (U) Secondary Missiles; (U) Laboratory Animals2S. TCINIOCAI O4U(CTIVE.* RI APPROACHI. IS. 000406" rP.a &MwfG.. P-.*g.. ,d-aga.E 6w .. S-. P_.. ,.. . .. -A * .0 c. ... u.ft

23. (U) To conduct a systematic evaluation of animal tissues (eyes and brains) sub-jected to trauma as a result of high velocity missiles striking the maxillofacial com-plex. The increasing numbers and velocity of missiles produced by modern weaponryresults in a significant increase "in the production of tooth and bone fragments whichcan do extensive damage as secondary missiles reote from the primary missile impactsite. The objective of this study is to provide a semi-quantitative evaluation of thesecondary effects of high velocity missile impacts and to provide data for the'IxxJuxter Man" system which will help to define treatment strategies for a wide rangeof combat wounds.

24. (U) Forty-five Texas Angora goats were shot in the maxillofacial complex with highvelocity missiles of various sizes in a USAIDR contract project. At autopsy the eyesand brains were removed intact. Detailed histologic evaluations of these tissues willbe made using conventional qualitative methods as well as new methods which will pro-vide quantitative data.

25. (U) (81 10 - 82 10) The eyes of 45 Texas Angora goats were prepared for histologicevaluation (H&E, PAS and Tricbram stains). The brains were examined grossly and xero-radiographically.- Ten of the goats sustained fractures of the orbit and one hadhemorrhage in the viterious body. Four of the brains had gross focal hemorrhage. Nogross or microscopic bone or tooth fragents were found in the brain or eyes. Becauseof their anatomic configurations the goats sustained minimal damage to eyes and brainsfrom mxillomndibular impact.

D- '*1498 pnOUG 0LOITIONS O" THISI ONM AN. OGS.LETE 30 6ONS I4..A 1NOV es

D, 'IN 8 0A" 46Q I MAN 04 A OR A*V USK) AMC 0SOL arTe

. . .. . • -- _mm m Rm -- r ss 15

P)JECXT: 3M161102BSIO

WORK UNIT TITLE: THE SECONDARY EFFECT OF HIGH VELOCITY MISSILES ONCRANIOFACIAL TISSUES

PRINCIPALINVESTIGATOR: LWI DAVID M. LEWIS, DC

An Evaluation of Eyes and Brain fran Laboratory Animals

Wounded in the Maxillofacial Ccmplex with Variable Moss

High Velocity Projectiles

PROBILM: To conduct a systematic evaluation of animal tissues (eyes andbrains) subjected to trauma as a result of high velocity missiles strikingthe maxillofacial complex. The increasing numbers and velocity of missilesproduced by modern weaponry results in a significant increase in theproduction of tooth and bone fragments which can do extensive damage assecondary missiles remote from the pcimary missile impact site. Theobjective of this study is to provide a semiquantitative evaluation of thesecondary effects of high velocity missile impacts and to provide data forthe "Computer Man" system which will help to define treatment strategiesfor a wide range of combat wounds.

APPROACH: Forty five Texas Angora goats were shot in the maxillofacialccmplex with high velocity missiles of various sizes in a USAIDR contractproject. At autopsy the eyes and brains were removed intact. Detailedhistologic evaluations of., these tissues will be made using conventionalqualitative methods as well as new methods which will provide which willprovide quantitative data.

RESULTS: All goats that did not succumb to missile impact or anestheticwere sacrificed 15 minutes after impact. At autopsy the eyes and brainswere removed intact and gross damage was noted. Focal gross brainhemrrhage was noted in four goats. Bony fractures of the base of theskull were found but no bone fragments or tooth fragments were found ongross or xeroradiographic examination of the goat brains. Xeroradiographictechniques were found to be superior to routine radiographs of the brains.

The eyes were prepared for histologic examination. Multiple sections weremade fran each eye and H&E, PAS and Trichrcme stain were done. Ten of thegoats had fractures of the orbital bones. Gross hemorrhage of the orbitalfat was noted in goat #22828. Microscoptc evidence of intra-ocular hemor-rhage was found in one goat #22851. That goat had focal mild to moderatehemorrhage of the vitreous body. The source of the blood was most likelythe retinal vessels and could have been related to ocular trauma. Otherfinding indicative of trauma but probably not related to the present testwere focal melanosis of the peripheral endothelial cell and filtrationangle, cataracts, keratitis, and osseous metaplasia of the posteriorsclera. With the exception of the vitreous body hemorrhage in one goat,all of the changes indicative of damage are chronic changes. Since the

16

goats were sacrificed 15 minutes after impact it must be assumed that thechanges noted were not a direct result of the present test. All goats hadartifactual retinal separation. No gross or microscopic evidence of bonefragment or tooth fragments were found in any of the eyes examined.

The elongated maxillofacial anatony minimizes the effect of secondarymissile generation (bone & tooth fragments) on the eyes and brains.Because of this anatomic variation the correlation of damage between thegoat and human maxillofacial complex is poor.

17

r I I acI Access 08I , 00m7,RP Co "O0"

RESEARCH AND TECHNOLOGY WORK UNIT SUMMARY DA OE 6022 82 10 01 DD-D & (A N)6 J

8b 1TC 10 .:C09 ACCI

81 10 01 D. CHANGE U NL 0E, DUO

to Ne./COOgs:I PROCRAN ELEM4NT PROJECT NUNGER TAIS AREA bUMImE W04K UIT NU.0eR

t POIWA" 62775A 3S162775A825 AB 014

Is . 62775A 3S162775A825 AC 014

I ,/ / STOG: 80-7.2:5fl TITLE (Pt4 at ,54.9t CP.*jflS q, C"1

(U) Preventive Dentistry Measures of Military Significance'I- SCI(NTIPC &*O TVCMOLOGIC.I ARMAS*

012900 Physiology; 002400 Bioengineering

71 01 CONT DA C. IN-HOUSEI7 CONTRACVORAMT * AtSOUMaCI ESVIMATC A. 1.TRSIW% U 1 I b s0 (I-. .. a

& OA*TEStEPDECTIVI Si AI m1'oa

.61. .... FIS 82 0.2 21

. .10 O .. I CUM. AMT. 83 0.2 5011 RESIPOOS40LE 000 ORGANIZATOW mPUO.G RAIAT0

.AO::* U.S. Army Institute of Dental Research ""'GU.S. Army Institute of Dental Research

.oo.aos*Washington, DC 20307 Aoo='SS- Washington, DC 20307

PRINCIPAL. -SSI A TO. If-.A IS-A 1 U S 5 -P *''~

REDoWS,.tS ASoLo -A-1 " LORTON, L.

SWEENEY, T.P. ,,LF.a.oa (415) 561-4845

,.. ..o. (202) 576-3484 oCI,., .CUM-.. ACCO ... 01. POC:DA

31 ASSOCIAVS -ItCTI03t GIaNSRA. al $l[Ae* E'GA~l

MADER, C.

FIC ___,1 K-04o1 ..... eAC .- .S. C ,..-.... C-I (U) Marginal Leakage; (U) Resin Restorative;

(U) Post and Core Restorations; (U) Dental Emergencies;TICMCAL O9J9CTVC 1 14 APPRAoCK IS I-OG-4IS 'r LAN&,.D...PRA. *-'s-. . P"C- ... . t - --5...* .

23. (U) To develop new and simplified methods of preventing disease related dental

emergencies in the field. To assess new methods of (1) improving the biologic

management of militarily relevant oral conditions and (2) improving the cost-

effectiveness factors of preventive dental therapy in the military.

24. (U) Studies will be conducted which will (1) develop and evaluate improvedmethods of dental care which will prevent dental emergencies in the field; (2)

develop more rapid and effective means of identifying and treating soldiers with

at-risk-profiles for field dental emergencies.

25. (U) (81 10 - 82 10) A cavosurface margin design in the placement of resin

restorations was developed which should decrease marginal leakage, one of the

primary causes of restoration failure and resulting dental emergencies. A combined

etching procedure and placement of a low viscqsity resin demonstrated good adapta-

tion to denin. This is a first step in developing a clinically satisfactorytechnique for resin post and core restorations. Improvement of such restorationscould reduce fieL.6 dental emergencies.

18

D, 14 9 8 A I I I f 1-1

4L m mmm m m m l

PROJECT: 3S162775A825

WORK UNIT TITLE: PREVENTIVE DENTISTRY MEASURES OF MILITARY SIGNIFICANCE

PRINCIPALINVESTIGATOR: LTC LEWJIS L. LORTON, DC

Evaluation of a New Restorative Procedure for Resin

Restorations Designed to Prevent Dental Emergencies on the Battlefield

PROBLEM: Marginal leakage is the major cause of failure of resin restora-tions and may result in a dental emergency. Thousands of resin restora-tions are placed each year by Army dentists. The development of a tech-nique for resin restorations, which would prevent marginal leakage, ishighly desirable as it would prevent the major cause of failure of thisccmmonly used restoration. In addition, it would save time expended by thedental staff and soldier/patient to replace resin restorations that haveprematurely failed. It would also prevent dental emergencies associatedwith restoration failures.

APPROACH: Cavity preparations with traditional and experimental cavo-surface margin designs were prepared in extracted human teeth. The marginswere etched with orthophosphoric acid (37%) for one minute and then filledwith several comnmercial dental resins. The teeth were dissolved withnitric acid and the cavosurface margins of the remaining, undissolved resinrestorations were examined with a scanning electron microscope to evaluatethe adaptation of the various resins to the traditional and experimentalcavity designs.

RESULTS: One low viscosity resin demonstrated superior adaptation to allexperimental cavosurface margin designs. This fine adaptation may preventmarginal leakage around resin restorations done with traditional techni-ques.

19


WORK UNIT TITLE: PREVENTIVE DENTISTRY MEASURES OF MILITARY SIGNIFICANCE

PRINCIPALINVESTIGATOR: LTC CARSON L. MADER, DC

Evaluation of a New Restorative Procedure for Post

and Core Restorations Designed to be More Economical and to

Prevent Dental Emergencies on the Battlefield

PROBLEM: Traditional methods for fabricating post and core restorationsare very time consuming and expensive as these techniques require sub-stantial amounts of time from the dentist, the dental assistant, thelaboratory technician and the soldier/patient. In addition, relativelylarge amounts of precious metals (usually gold or a gold alloy) are re-quired. Also, forceful traumatic blows to anterior teeth with cast metalpost and core restorations frequently result in fracture of the root. Thiscan easily cause a painful dental emergency requiring inmediate removal ofthe fractured tooth and result in a soldier being lost from duty. Themissing anterior tooth must also be eventually replaced. This requiresanother, multi-step procedure involving additional expense and time ex-penditure by the dental staff and soldier/patient. The development of asingle-step technique for the fabrication of resin post and core restora-tions could save the Army considerable expense and time expended by thetreating dental staff and the soldier/patient. In addition, resin post andcore restorations could prevent dental emergencies caused by traumaticallyinduced root fractures associated with traditional cast metal post and corerestorations.

APPROACH: Single-rooted, extracted human teeth were treated with conven-tional endodontic techniques to receive traditional post and core restora-tions. The canals- were then etched with acid and filled with a lowviscosity resin. The teeth were longitudinally split in half and examinedwith a scanning microscope to evaluate the degree of resin penetration intothe p -ent dentin tubules. Reliable and consistent resin penetration otthe patent dentin tubules may be the first step in development of a new,one-step, resin technique for fabricating post and core restoration whichwill be faster, cheaper, and prevent dental emergencies.

RESULTS: Scanning electron microscopic examinations of the canal walls andthe resin cores which had formed the canal wall-resin core interface beforethe teeth were split, showed that the low viscosity resin did effectivelypenetrate the patent dentin tubules.

20

I. AGENCY ACCSUS D.ATEOF Su"ARV. I aa coopino. from

RESEARCH AND TECHNOLOGY WORK UNIT SUMMARY DA 0G 60331 824 100 D-OR&E(AR)6.06

.OATS PHOY SUFY 14. NINO Of SUMMARY L. SUMMARY ;;7 " REUIT"7'SORAUS"i Afer OW VN SIRU *S . augfl OA0,EVLO SUN______________________________ O TRCTO ACCESS

8110 01 Do CH{ANGE jU.. NL 00 1r5 0NO.I A.WGff1. MO.'ES ROGAM EN RJECT "U."ER TASK ARECA 1UM00164" UNIT "uMOERf

6=75A 3S162775A825 AB 002b OTVOT11 627A3S162775A825 AC 002

b. ~ 6275A / / SLG 80-7.2:5_____111. TITL.E (Po~md Irl SoONNHP C168*0REe Cod~

(U) Development and Evaluation of Dental Materials and Materiel for Army UseI& SCIENTIFIC ARO TECNUOS.00SCAL ARC"*

010300 Miscellaneous MaterialsILSATDT 1. SYINMATES COMPLETION SATE IS ruUWIN6 AGENCY S PESoRMAWCt 1111114

69 01 . WN'r DA I IC. IN-l!XJSIT. CONTRACT/661ANTme RCK 9IAC a1"FSM L"A va %Irm0towwooN

IL 0ATgS/EPPECTIVS. IX04RATOON:

IL NUmeaw ISC:A2L 1.0 75A TVPE A AampuUT 'aEA. .. .

A6 EINft@P AVIA*Sk L.CUM. AMT. 831.5 850. 043141 NE01100 W ORSANIZAYION 11- P901 P-01 ORGAISATIOW

*AM*USA Institute of Dental Research WM USA Institute of Dental Research4AOOEU.* Washington, DC 20012 ,,stess.-Washington, DC 20012

r PRINCIPAL IWVESTIGA.TO Ifoood MAP Of W S. 4oftom.I

SIUMANS661.9 ICHVIUAL A., Vennidlyea, S.G., L7C, DCNASIE: Sweney, T.P., 00L, DC TELEPHONE- 415-752-8343TELEPH"OSIE 20-576-3484 SOCIAL SECURITT ACCOUNT "UNGER POC :DA

GE.NER4SAL USE UCAC4VSIA01

Flreign Intelligence Considered NAme, Woolsey, Go . S.C D4XK9VQO6(~&.SAK INS.AHr ~dJ UJStorage Stabilfy;(U) caoste estoratives;

Criteria for selection of mterials, devices or techniques for evaluation are based onanticipated high potentials for: (1) savings of fiscal and/or manpower resources; (2)wrk siaplificat ion; (3) improved health care delivery in combtat areas; and (4) enhancesafety with respect to professional and ancillary personnel as well as to the patient.

24. (U) New mterials will be evaluated on the basis of the following parameters: Cwl-position, microstruxcture, physic-al and mechanical properties, cytotoxicity, and clinicalperformance.

* 25. (U) (81 10 - 82 10) The storage stability of 5 caiposite restoratives and 9 imp~ress-ion materials were subjected to cyclic teperature-humidity conditions for 5 mornths.On~e caqposite lost tensile strength and one lost setting characteristics. All theimpression materials showed some changes. Continued wor with carbon fiber reinforcedacrylic resin indicates potential use for the.stabilization -ot nmxzillofacial fracturesor field repair of prostheses. A method of identifying potential corrosion problem~ inbiainedical alloys used by the Army is being developed and is near complet ion. 'TheI development of a procedure to improve porcelain-to-metal bonds is in progress. Somesuccess has been obtained to date. Potential savings in fiscal and manpoer expendi-tures that could result from this latter effort are considerable.

21

uu-11#u. 00INI6 _OMM 6. 615

P*VIUS COITION$ 0: TIS FORM ARE OSICLE YE 00 1 ORMS 1408A 1NO0V 6SD Dv~sAON"198 A0la1*S lASI I MASS 45 1PaR ARMY USE I ARI OSOL EYEE

PROJECT: 35162775A825

WORK UNIT TITLE: DEVELOPMENT AND EVAILUATION OF DENTAL MATERIALS ANDMATERIEL FOR ARMY USE.

PRINCIPALINVESTIGATOR: LTC S. G. VERMILYEA, DC

Storage Stability of Dental Materials of Interest to the

Military Dentist

PROBLE4: Logistical burdens inherent to the world wide deployment ofmilitary dentists are manifold. Among the most prcninent problems arethose related to the shipment and storage of certain restorative materials.For example, organoinorganic ccmuosites, waxes and elastcmeric impressionmaterials which can be employed successfully throughout the North Americancontinent may not survive shipment to less temperate regions. In addition,the extent to which prolonged storage under tropical, arid or arctic-likeconditions might affect the properties and characteristics of thesematerials is unknown. At the present time there is no rapid "aging" testto assess the potential storage characteristics of materials.

APPROACH: A programmable temperature/humidity chamber was used to simulateextremes of tropical, subtropical and arctic-like climatic conditions.Unopened, "as received" packages of selected materials will be exposed toeach storage condition of periods ranging from two to six months. At thetermination of each storage period the materials were subjected to standardtests to assess their physical and mechanical properties and manipulativecharacteristics. The features of the stored materials were ccmpared tothose of the "as received' materials.

RESULTS: In this initial portion of the study, five ccmposite restorativesand nine impression materials were subjected to cyclic temperature/humidityconditions. When compared to the base line values the capressive strengthvalues for the caiposite restoratives were not significantly altered byexposure to a tesperature/humidity cycle (-10" to 46"C and 50-89% respec-tively) for five months. Tensile (diametral compressive) strength of onematerial (Adaptic) decreased more than 50% subsequent to the same cycle.In addition, one material (Profile) exhibited a slight deterioration of itssetting characteristics after five months. With the impression materials,exposure to the temperature/humidity cycle elicited a general increase inpermanent deformation for the polysulfides and one of the vinylpoly-siloxanes. Increased 24 hour dimensional change was observed with all ofthe materials except one vinylpolysiloxame and the polyether product. Theproperties of the polyether material were not altered by the temperature-humidity cycle. It should be noted that the observed changes in thematerials are measurable in the laboratory but may not be of great clinicalsignificance. Additional work is needed to alter the temperature/humidityprogram to (a) more closely duplicate expected field conditions (b) shortenthe time required to produce property changes and (c) correlate the accele-rated aging program with the expected shelf life as defined by the productmanufacturer.

22


WORK UNIT TITLE: DEVELOPMENT AND EVAiUATION OF DENTAL MATERIALS ANDMATERIEL FOR ARY USE

PRINCIPAL

INVESTIGATOR: LTC S. G. VERMILYEA, DC

Effects of Fiber Reinforcement on the Dynamic

Properties of Denture Base Resins

PROBLEM: Flexural fatigue fracture of acrylic denture bases are notuncommon occurrences. Solution to this problem often requires the con-struction of cast metal bases; a difficult, relatively expensive and timeconsuming task. Increases in the flexural fatigue resistance of acrylicdenture bases would reduce the need for metal based dentures, allow thinnerprostheses to be used in conjunction with soft lining materials and permitthe use of thinner yet rigid stents during intraoral stablization offactures.

APPROACH: Silanized and unsilanized carbon fibers were processed longi-tudinally, perpendicularly, in a criss-cross pattern, as well as randcmlyto the long axis of acrylic specimens. The specimens were machined to asize of 60m X 10m X 2.5im and subjected to three point cyclic loading ata rate of 325 cycles per second under a 5kg load until failure occurred.Additional specimens were subjected to transverse and tensile strength de-terminations.

RESULTS: The data indicate that a 100% increase in fatigue strength andtransverse strength is possible with silanized longitudinal fibers placedin the center of the specimen. However, this is a technically difficulttask. The use of 0.4% by weight of randamly dispersed chopped (3-4nmlength) fibers resulted in an 80% increase in fatigue strength withoutadversley affecting the other properties. It is conceivable that theincorporation of chopped fibers into prepackaged denture repair materialswould also provide similar property increases at minimal cost. At thistime this project is complete.

23

. mm dlmm i~m mmm m L


WORK UNIT TITLE: DEVEIOpMENT AND EVALUATION OF DENTAL MATERIALS ANDMATERIEL FOR AF4Y USE

PRINCIPALINVESTIGATOR: LTIC S. G. VERMILYEA, DC

Electrochemical Profiles of Dental Alloys

PROBLEM: World wide deployment of biomedical materials and materielsubject these items to extreme conditions and increase their potential fortarnish and corrosion. Currently, there is no rapid, reliable method forthe prediction of the potential corrosion resistance of equipment andsupplies in a given environment nor the potential useful life of manymetallic supplies. Furthermore, many bicnedical alloys, when placed incontact with living tissue undergo dissolution with the subsequent releaseof ion species potentially toxic to the host tissues. A method of identi-fication of the potential corrosion resistance of biomedical materials andmateriel would be invaluable to the rational selection of the most stableand safest items for use by the U.S. Army Health Care Delivery System.

APPR1 CH: Initially, a study will be undertaken to determine appropriatespecimen geometry and finishing technique to yield reproducible results.Subsequently, potentiodynamic polarization, linear polarization (corrosionrate determination) and galvanic coupling procedures will be combined withformed film analysis to determine the species of ions released and thepotential corrosion resistance of the test materials.

RESULTS: A specimen geometry and metal finishing procedure has beendeveloped which appears to produce consistent results. Briefly, cast discspecimens are mounted in a thermplastic medium and polished metallographi-cally with 0.3 micrcmeter alumina. Subsequent to each electrochemical de-termination, the specimens were repolished with 0.3 micrometer alumina.The use of the technique with several base metal alloys has given repro-ducible results. The remainder of electrochemical techniques will beinitiated upon receipt of the necessary instrumentation on the West Coast.

24


WORK UNIT TITLE: DEVELOPMEqT AND EVALUATION OF DENTAL MATERIALS ANDMATERIEL FOR AR4Y USE


Effects of Alloy Surface Treatment on the

Porcelain-Metal Bond

PROL1: Ceramo-metal restorations account for a significant portion ofthe fiscal expenditures within the Army Health Care Delivery System. Therapidly fluctuating price of gold and other noble metals has pramptedintense interest in the use of less costly base-metal alloys as alternativeto the high noble content materials. Savings of 80-90% in current alloyexpenditures are possible with the use of base-metal materials. Unfortu-nately, a significant draw back to the use of these materials has been thefrequent failure of the porcelain-alloy bond. Such failure requiresrefabrication of restorations with conccmitant additional expenditure offiscal and manpower resources. The mode of faiure (i.e., cohesive withinthe metal oxide layer or adhesive at the oxide-metal phase boundry) suggestthe possibility of modifying the substrate metal surface prior to porcelainapplication to improve the integrity and strength of the porcelain-alloybond.

APPRO)ACH: Selected base metal alloys and proprietary coating agents wereused in a randamized desigp to determine the effects of the coating agenton the strength of the ptrcelain-metal bond. Specimens were 1/4 X 1/16inch cast discs to which porcelain was applied in the shape of a cylinder.The porcelain cylinder was embedded in a tray acrylic and the disc sub-jected to diametral loading at a cross head descent rate of 0.02 inch perminute. The apparent porcelain alloy bond strength was calculated on thebasis of the apparent area of porcelain alloy contact and the load at shearfailure.

RESULTS: With two of the alloys, the use of a metal conditioning orcoating agent prior to porcelain application had no effect on the apparentbond strength over that observed with uncoated specimens. With one alloy,the use of the bonding agent resulted in a decrease in apparent bondstrength as ccuiared to specimens in which the coating agent was not used,whereas the remaining alloy exhibited the similar bond strength values withany of the coating agents. Examination-of the failure sites indicate ametal-oxide or oxide-oxide type of failure. Further work is in progress todevelop new finishing procedures, firing cycles and a proprietory "bondingagent" to overcome the difficulty in obtaining a reliable porcelain-alloybond with the base metals. In addition, the effects of different porce-lains with the various alloys will be assessed.

25

6. a*~cMC ACCESSION A.D o f 0 suWUAn alle ar UO scoo o ., I

RESEARCH AND TECHNOLOGY WORK UNIT SUGARY Di OG 6034 .82 10 01 DD-.DR&,(AR)j6

& ATI P0MV 4. PIUS @ WUMANY S IMMAPY ICTI 1.P jOPS SUINZT WA~OU iA WO I6ITp*w SUMR peSCCP Os. " O L P, "CR

81 10 01 D. CIE ;::CA-K

to. NO./cOOC8,4 PIOGAM GLEMUNT PROJECT NUNDCA TAIN AREA NUNOIA WORN UNIT NUMISCR

•A "M .. 62775A 3S162775A825 AB 003b. CO~mui2o775A 3S162775A825 AC 003V JW10 lll l I/ Y SOG 80-7.2 5

11a. TITLC rP.... .40 *.N Ir .U-' CCd

(U) Development and Improvement of Metallic Restorative MaterialsI& SCICKTIFIC AND T(CNMMOOICAL AMCAS'

009900 etallurgy and MetallographyIS. STAN? DATC K0 STIMATCS COMPLEION DATE It. FU.01MG *SICC 0 (VUAcK.T69 01 j CflT DA C. IN-MJUSE

IV. €ONlTRACTSGMANY W- RESOURCES ESTIMATE I A PM0OWlCSO.L MAN INS Is FU-011 (ff M- )

IL DATEV0 CTIVC8 RXPIRATIOUN .. eC1 1

b .uneen: FISCAL 82 1.0 544, TVPVC A 450gWT YCAM tTVWFUUY_

111101o AWARD C.CUU. A.. 83 1.0 65IS. UUOUUDL.S 000 044ANO RATION JR. .gmVOmwe ORGANIZATION

"&"* USA Institute of Dental Research MA,* USA Institute of Dental Research

A oae. Washington, DC 20012 Aa,,,. Washington, DC 20012

PRINCIPAL INVCSTIGATO. j1-- MAN -' U.. aA"A ..... lm

IMSPOO IDIVISIAL .M:* Vermilyea, S..-- I Sweeney, T.P. ,,l,.Pul: 415-752-8343TLOt,: 202-576-3484 SOCIAL SECURITY ACCOUNT UOE POC :DAI1 GENEPAL USE AOSOCIAT( IVIRSTIGATONS

Fbreign Intelligence Considered .AMC Woolsey, G.._,_ Kuffler. M.

MR.11040 (I ....EAC N-11"I ..... &*' (U) Base Metal Alloys; (U) Casting Accuracy; (U) Solder-

in& Base etals; (U) Investment Techniques.28. TCCUICAL OGJCCT$vE., 29L APPROACH. A& P*OGAEIS PS.WA 1-0. j-ARAR* 04-104 2, .4 P-.,o ... .1 .-. -0 .- -a.,-cd.

23. (U) Annual Army expendifures for precious metals utilized in the fabrication offixed dental prostheses are in the vicinity of $1,000,000. The cost of an equal volumeof base metal alloy is $30,000. Properties of base metal alloys indicate however thatthese alloys cannot be utilized for small castings without drastic metallurgical modi-fications. This work is therefore being conducted to (1) develop heat treatment methfor controlling properties of nickel-chrcmium based casting alloys; (b) evaluate nickelchromium based alloys for use in operative dentistry.

24. (U) The properties of nickel-chromium based alloys will be studied in detail byvarious physical methods available in order to devise procedures which will optimizetheir usefulness. Any improvement obtained will be evaluated clinically.

25. (U) (81 10 - 82 10) Reliable methods for casting base-metal alloys as replacementsfor noble metals have been lacking. A study ccmpleted on 6 comercially available basemetal alloys showed that the casting accuraty of 5 alloys could be dramatically im-proved by modifying investment techniques. The techniques developed are reliable andsimply applied. The resulting savings in money and manpower should be significant.Attempts to develop reliable techniques for soldering base-metal alloys have thus farnot been successful. Studies are continuing.

26D "PR198 .VIOUS LOITIONS OP T~iS FORMAR OSS 0C0.E' TI 30OROAO 45* ... OV 55

WUARm ANO t450 I I M ARN TiIOR &AY USEI ARE OBSOLETE


UNIT WORK TITLE: DEVELOPMENT AND IMPROVEMENT OF METALLIC RESTORATIVEMATERIALS


Development of Techniques to Improve the

Casting Accuracy of Base-Metal Alloys

PROBLE4: Although the potential fiscal savings to be realized through theuse of base metal alloys in military dental practice is great, severalproblem need to be addressed prior to the institution of their routine.Among the problem areas is the development of methods for the routineaccurate casting of these alloys by personnel of varying skill levels.

APPROACH: Comimrcially available investment materials were employed inmodified investing techniques to construct full coverage restorations.Investment materials included Hi-Temp 2 and Ceramigold 2. Modifications toinvesting techniques were hydroscopic methods, alteration of special liquidconcentrations (Hi-Temp) and the use of Hi-Temp and Ceramigold specialliquid at selected dilutions with Hi-Tenp 2 and Ceramigold 2 investments.Two hundred twenty two wax patterns were constructed on individual stonedies, invested and cast in Unibond, Ceramalloy II, Biocast and Neobond IIalloy. After divestment each casting was seated on its die and examinedfor size and marginal integrity with the aid of examining loupes. Castingswere judged as: adequate (.); oversize (+); or under size (-).

RESULTS: When casting size was the determining factor the scores for thetotal sample distribution were: (+) - 49%; (0) - 32% (-) - 29%. Whenmarginal integrity was considered the scores were: (+) - 21%; (0) - 22%;(-) - 57%. Investment manufacturer's with 62% special liquid in a hydro-scopic technique produced consistently well-fitting castings with Unibond,Ceramalloy II and Biocast. In addition, 100% and 83% of the respectivecastings of Unibond and Biocast from Ceramigold 2 molds using 23% specialliquid and a non-hydroscopic technique were adequate. None of the techni-que modifications produced consistently well-fitting castings from NeobondII alloy. The findings suggest that investing techniques must be tailoredto the characteristics of the individual alloy.

27

• .i. a -- uu lnlm IN m ~ lnnlnln m ~ii m n L

P1aJECT: 3S162775A825

WORK UNIT TITLE: DEVELOPMENT AND IMPROVEMENT OF METALLIC RESTORATIVEMATERIALS


Tissue Response to Base-Metal Crown and Bridge Alloys

PROBLEM: The military is under constant pressure to reduce fiscal expendi-tures for noble alloys. In addition, laboratories and purchasing agentsare inundated by alloy manufacturers to purchase a particular product andmust do so if a "low bid" is received from a particular manufacturer.Alloys of seemingly similar ccr%)osition differ markedly in their potentialhistotoxic reactions. It would appear that the severity of any tissuereaction depends upon the cytotoxic potentials of ions made availablethrough dissolution of products produced by wear and corrosion of thealloy's surface. Since, in the military, the dental population is "cap-tive" and has little choice as to the type of alloy enployed, it is impor-tant that alloys of known safety and efficacy be used in the MilitaryHealth Care Delivery System.

APPROACH: Powders obtained from the wet-milling of five alloys will bepressed into tablets, sterilized and implanted subcutaneously in rats.Specimens will be harvested at 1 week, 1 month, 2 months, 6 months, and oneyear. The tissues will be processed and graded as to the severity ofinflammatory response.

RESULTS: To date, powders of two of the alloys have been produced. Addi-tional preparation will commnee upon receipt of appropriate equipment.

PRJECr: 3S162775A825

WORK UNIT TITLE: DEVELOPMENT AND IMPROV4ENr OF METALLIC RESTORATIVEMATERIALS

PRINCIPALINVESTIGATOR: LTC S. G. VER4ILYEA, DC

Development of Techniques for the Soldering of Base-Metal Alloys

PROBLEM: The potential fiscal savings to the U.S. Army Health Care De-livery System afforded by the use of base-metal alloys in lieu of expensivenoble materials has been well established. Heretofore, technology for theroutine accurate casting of many of these alloys has been developed withinthis laboratory. However, many times an intraoral dental prosthesis mustbe joined by soldering prior to ccpletion. Controversy exists as to themost appropriate soldering method and the degree of success to be expected.Development of a predictably successful soldering technique for use withbase-metal alloys will permit full realization of the potential fiscalsavings afforded by the use of base-metal alloys in Army Area DentalLaboratories.

APPROACH: Initially five base-metal alloys and five solders were employedin a randomized design to assess the effects of. each solder/alloy combi-nation on the tensile strengths of the resultant joints. Specimens werehalf-tensile bars fabricated from the substrate alloys by conventionaldental laboratory procedures. Soldering of each alloy/solder combinationwas acccmplished in accordance with the solder manufacturer's instructions.Soldered bars were subjected to tensile loading on a testing machine at acrosshead speed of 0.02' inch per minute. Strength and rigidity of thespcimens were assessed by analysis of the resultant load deflection curves.

RESULTS: The results indicate that the successful soldering of base-metalalloys is variable at best. Fully 38% of the specimens fractured prior totesting. Of the successfully tested specimens, tensile strength valuesranged from 4,500psi alloy with Unibond Pre-Solder. Precision of the datawas low with coefficients of variation ranging from 1 to 54%. In general,the ability to produce consistently strong contiguous solder joints fromany alloy/solder combination was poor. Surface and subsurface porosity wasa characteristic feature of the solder joints. Studies are now in progressto develop pre-soldering finishing procedures and alternative fluxingagents to improve the wetting characterstics of the solder on the substratealloy and hopefully the strength and integrity of the solder joint.

29

RESEARCH AND TECHNOLOGY WORK UNIT SUMMARY DA (Xi 6679 8s2; 10 01 OD-DR&E(AR)636

06 ISAYS *Rev sua IMS 0" SsIuA"' a. FUUA -=sl * AE~Y 9 EUSM . car" loillror I SOCCIFIC DATA ' LgVCL 00 SUN

81 10 01 D. 0N U~ N 104UIIs. Mo.icooes'$ PACORAM RLIKS111" P010jaCY RUNDE" YASE Afi MUMDt wot UNIT Mmen

0. PWAR F6775A 3S162775A82500b. CeaOmeuyTuo 62775A 3S!62775A82500

Fly Nt-ft / /1// / SIDG 80-7. 2:5

U)The Initial Treatment of Combat WoundsIL SCICATIFIC AND 1 1CUMOLOGICAL ARMAS'

012600 Pharmacology 012900 Physiology 002300 BiochenistrIL. STAN? DATE JI.(104tDCWSIMDTE IS, FW.NO AOESCY IS. PERF ASCE ofyUOO

81 01 Cxtr D e.]_ Sit. ONTACT/0MAT 1. 0IgSOUMCES ESTINAYE A. PMOPESZIM L NAM VSIA NS. (I. A -.A)

A. DATC&AFUPECTIVE 41114MATION;.VCD.

a .Vy1,44 A ANOUNTTA 0n DI

M, RIMDOO AWARO. I. CUML. AMT?.83109

US. ft"EI*MSSU 000*"MANSIATBOON _ = . 119a 1-"6 041411 @R A TWO I

"A"I USA Institute of Dental Research MAC, USA Institute of Dental Research

^90w Washington, DC 20012 AocoRESA Washington, D)C 20012

PRINCIPAL INVIESTIGATOR eP.M AN Itn U uS. A-*(t 91IIf""

WESPOUSISLE INDIVIDUIAL MNG!" Wynkoop, J."MAL aveney, T.P. TCLCewaic 202-576-3393

TCC-z202-576-3484 SOCIAL SECURITT ACCOUNT mumagER 1PX:DAIt. ggEtERAL USE ASSOCIArEI IN.VESTIGATORS

Fbreign Intelligence Considered -oel Setterstrni, J... 4V in c e n t , M i l l e r , R . -AX KYWODS P-A SAC IFI SOM*f Cll-df" .4 (U) Cellulose Triacetate; (U) Gentamicin; (U) CopolymerBandage; (U) Idocaine; (U) Laboratory Animals

IS. TCIIIAL OBJECIVE.' 84. A4PROACII. I. PROGRESS4 ,-uM16 gPn.Ss p...p66* 1dI 6, p--f ".8. e( Il I M S-Ift CI*IS1*(1ll CL')

23. (U) To develop a mmftipurpese wound dressing which will provide anesthesia, antisep-sis and hemostasis so that, where appropriate, the result will be rapid return of thewovunded soldier to duty as well as reduction of the rmrbidity occasioned by delayeddefinitive treatment and secondary complications.

24. (U) Cotract developed drug release systm will be evaluated in animal miodelsdeveloped specifically for that purpose. Various methods and nmterials for mintainingcontact over a variety of rmxillofacial contours will be evaluated for their utility ofapplication and use in the combat situation.

25. (U) (81 10 - 82 10) A biodegradable polylactic-pollyglycholic acid copolymer fabricmatrix containing the anesthetic lidocaine was evaluated in a rabbit model for itsability to provide an extended analgesic effect. Based on tac-tile responses in theexperimental animl it was demnxstrated that a wound covering can be produced whichwill deliver appropriate drug levels over a 2f hour period. A new cellulose triacetateaterial is being evaluated as an antibiotic delivery systemi for incorporation intobandages. In vitro tests indicate the nmterial is a good slow-release medium for bac-tericidal agents. A wound model has been established in guinea pigs and a cellulosetriacetate-gentanicin systemi is being evaluated on these wunds for its ability to treat

S.aureua infen-tions.

30

0RiOU ffo'Fam 0OTOSO TmllS FORM AlRe OGSC.LY .30 PORMS 1494A I NOV 65D0,Weoft 14 98 .aS1 4096 1 MAR of ':O on4. Afu SIE1 &or OUSOLETE


WORK UNIT TITLE: THE INITIAL TREATMENT OF COMBAT WOUNDS

PRINCIPALINVESTIGATOR: CPT JUDSON R. WYNKOOP, DC

Developiment of a Multipurpose Wound

Dressing for the Soldier

PROBLEM: Despite advances made during the past thirty years in ourunderstanding of the healing process, the lack of an adequate wounddressing or bandage persists. There is a need for a multipurpose wounddressing, adaptable to combat situation, which will provide anesthesia,antisepsis, and hemostasis.

APPROACH: A biodegradable fabric will be used as a matrix for extendedrelease anesthetic and antiseptic products. Anesthetic and antisepticmaterials will be evaluated prior to combination with the matrix. Serumlevels of anesthetic will be measured at 0, 10, 30 minutes and at 1, 2, 4,6, 10 and 24 hours following injection of extended release materials andcompared to serum levels following injection of standard as well as coamer-cial local anesthetic solutions. The tactile responses of animals will bemeasured following injection of standard and camercial solutions and com-pared with the tactile responses following injection of extended releaseanesthetics. A separate model system, using rats, will be developed tomeasure the antiseptic quality of extended release products.

RESULTS: Preliminary data indicate that an extended release local anes-hTetic can be manufactured which will deliver measurable quantities over a24-hour period. Further evaluation of products manufactured under contractis in progress and should demonstrate the maximum serum levels obtainablewith slow release local anesthetics.

31

• - .rm. -- -- m mm lmmmmmm ~m mm! m mm mmllm lllm1

PRO=: 3S162775A825

WORK UNIT TITLE: THE INITIAL TREATMENT OF COMBAT WOUNDS

PRINCIPALINVESTIGATOR: JEAN A. SETTERSTROM, PhD

Investigation of a New Cellulose

Triacetate Film As A Wound Dressing

PROBLEM: Wound dressing technology has not advanced in recent years at apace consistent with the technology available for its improvement. Minorwounds in mass casualty situations tend to receive little attention. How-ever, wounds which do not initially prevent the soldiers from performinghis duties are often the occasion for infections which ultimately result insignificant lost duty time. Wound dressings are required which not onlyprovide hemostasis but also relatively long term prevention or eliminationof infections. This would reduce treatment time, morbidity and performancedecrement.

APPROACH: Extended drug release technology offers a solution to the pre-vention or elimination of infections in minor wounds through the incorpora-tion and zero-order release of medicaments frcm appropriately designedwound dressing materials. A new cellulose triacetate film with an extreme-ly high porosity is being studied for its ability to release antibioticsover an extended period into standardized wounds in guinea pigs infectedwith known levels of Staphylococcus aureus. This wound model will be usedto determine the release kinetics and anti-infection capabilities of filmsloaded at various levels with different antibiotics.

RESULTS: A wound model has been devised that consists of surgicallycreated one inch square, full thickness wounds on the backs of guinea 6gs.Each wound is contaminated with a broth suspension containing 2.5X10 S.aureus. Wounds are covered with the cellulose triacetate film and at 3days post-wounding the film is removed and the wound is scrubbed with abuffer-detergent mixture and the wash is assayed for bacterial content.Tissue is also remved from each wound site and assayed for bacterial con-tent. Under those conditions u~ing unloaded cellulose triacetate film amean bacterisl level of 6.5X10 is assayed in the wash solutions and inexcess of 10 organisms/gm in the tissues assayed. Cellulose triacetatefilms are currently being drepared under contract. Films with variousloadings of gentamicin and possibly clindamicin will be evaluated in theabove animal model.

32

I *flkC1 *CC9SSIO . I t DAf OF IU*A4' tapopr CO OL MMOLRESEARCH AND TECHNOLOGY WORK UNIT SUMARY DA 0G 8670 82 10 01 DDDR&E'AR)6J6

T a.SUMARY CT; V CCUAT 1 atRAO401 II& a." INSYrN. Ak syec(CI IC OAT. - . 19VtL :1 SUNtew 0 ACTO ACCESS81 10 01 D. CHANGE U 0. 0 Ubgy

I*. wo /COOam: •

01ROGRIAIM ErLCUSNT PR~oA~CT "NIVIV[R TASK AREA NIUI/NIR wongK UNIT tU.&CM~ea "POINT 62775A 3S162775A825 AB 011

IL C., b..TI.. 62775A 3S162775A825 AC all- ,OIt€11 1111111111114 SI1CG 80-7.2:5 .:It. ? ,,-L--A. I,.- CA.... C--R" (U) Development and Evaluation of Dental Materiel for Field

Use.IL SCICTIFIC ANO TCCNNOLOGiCAL AQ9A$*

002400 Bioengineering 010300 Miscellaneous MaterialsI* AN IIIATE a E?;ATEO CMPLETIOR OATE ISL 'U014NG AGINCY 0I. PIrOMANC[ (TkOO

CONT DA I C. IN-BDUSEIi. OITNACTIG-A T I Al[sUNCEs ISTIMATC & PNOFC6SIONAL MAN RIS h U 0N (1 v-N _)

* OAVOER IFPEC TI VEI LIR $QATIOR:im[

k"U"S " FISCAL 82 0,.5 165IA TYPL: A.Ov' 'CANNV

ft IMO AMINO. I. CUM. AMY. 83 2.0 165It. ft"PGNUS.E oca DO N AAU OR 1 O 30. PEORORWO0GANIIZATIONI

-- *t USA Institute of Dental Researcl. -:" USA Institute of Dental Research

AtooNam Washington, DC 20012 A"°* Washington, DC 20012

PRINCIPAL INvETIGATOR , AashinII IU X. aA" 2I0012

,ESPOSISA.E IN-WOVOAL N*ME: lorton, L. , LTC, DC-ANI 9%eeney, T.P. ,,l,9.0m,: 415-561-3162• ,L,0 ,, 202-576-3484 SOCIAL SECURY. ACCOUNT NU*E ,C :DAAt A[m11AL VS AtSOCIAVC INVCSTIGATOAS,

Foreign Intelligence Considered .. Vandre, R. RHM, H.NAME-: Grover, P.

X X.0(P--& EAC. -0NIR C1" ........ ' (U) Dental X-ray; (U) Dental Cutting Instrument; (U) Sub-traction Radiography; (U) Radionuclide X-ray System: (U) Laboratory Animals

82 T9CNNICAL OGJECYVt.* 24 APPRIOACM. JL 1.RO0R111 r(4"" v-A. -0.S . 'IO-WIUI. 61 .. a . p.... -,R . .. A h . . -1, CI.Iei~ CN.;

23. (U) To assist in the development of dental equipment capable of reliable perfornmncEand easy maintenance under all field operational conditions. Included are the develop-ment of concepts for field dental equipment which is miniaturized, lightweight, energyefficient and low cost.

24. (U) Conceptual and basic engineering requirenents for a field dental x-ray systemand a field dental cutting instrument will be studied. Current technology will bereviewed for its ability to produce the needed design criteria and advanced technologyrequirements will be identified. Experimental devices will be constructed.

25. (U) (81 10 - 82 10) -tudies have been initiated on the development for field use of(1) a hand-held, battery-operated, real-time dental x-ray system; (2) a radionuclidebased dental x-ray system, and (3) a field dental cutting instrment based on fluidicprinciples. A 'bread board" lightweight, battery-operated, real-time x-ray has beenevaluated clinically and found to produce excellent real-time imaging. Prototype de-signs have been submitted for construction under contract. The application of computer-ized subtraction radiography as a means of improving the detection of hard tissue woundsis being evaluated in vivo. Osseous wounds have been created in dog mandibles and thedata are being evaluated.

33

.z'..1498 XiO 14WS1 MAN AAFOARMY l ..... OSOLETE .. MS ARSA [OV

PRaJECT: 3S162775A825

WORK UNIT TITLE: DvELOpMENT AND EVALUATION OF DENTAL MATERIEL FOR FIELDUSE

PRINCIPALINVESTIGATOR: LTC LEWIS L. LORIXN, DC

A Dental X-Ray System for Field Use

PR OBLEM: The current field dental x-ray apparatus is heavy, bulky, and hasa high energy consumption. The dependence on conventional film and devel-oping is also inefficient in a field situation. New concepts are requiredfor producing dental x-rays in the field which are miniaturized, light-weight, energy efficient and cost effective. The energy requirement is aparticularly pressing problem inasmuch as an ever-increasing demand forenergy must be met for the wide variety of energy-intensive equipment usedin the field. Self-contained equipment with independent energy sourcesmust be developed if maximum utility and flexibility are to be maintained.Likewise, weight and cubic displacement are important to withstand therigors of the field with minimal maintenance and down-time.

APPROACH: Two concepts are being evaluated with respect to energy sources.One is a battery operated system; the second is a radionuclide drivensystem. Current technology is being evaluated and correlated with userinterface to determine if real-time imaging or a dry film (e.g. Polaroid)approach are sufficient for the diagnostic requirements of the field.Current technology is also being evaluated for miniaturization and weightreduction potential.

RESULTS: A real-time x-ray device utilizing an advanced battery system, agadolinium oxy-sulfide x-ray detector screen and a small x-ray sourceoperating at 40kvp was evaluated in a "bread-board" configuration for itsability to visualize endodontic procedures performed in monkeys. A totalarea of approximately 1 square inch could be seen with sufficient clarityto accurately determine the results of the root canal procedures. Subjec-tive evaluation by a number of dentists indicated that the level of diag-nostic information obtained was good to excellent. The total systemweighed just under 20 lbs. A second system currently in the design stageis based on the use of a radionuclide as an energy source. A mock-up x-raydevice has been constructed which weighs less than 1 lb and has a cubicvolume of less than 27 cubic inches. The energy source proposed for thisdevice is a gadolinium nuclide with a useful half-life of approximately oneyear. The source will be contained in S tantalum shield with a "dead-man"switching device for producing the x-ray exposures. Exposures will berecorded on extremely fast Polaroid film. All calculations have been com-pleted and the test sources are being ordered.

34


WORK UNIT TITLE: DEVELOPMENT AND EVALUATION OF DENTAL MATERIEL FOR FIELDUSE

PRINCIPALINVESTIGATOR: LTC LEWIS L. LORTON, DC

Development of a Field Dental Cutting System

PROBLEM: Currently available field dental units, although considerablyreduced in size and weight compared to fixed installations, are stillamenable to further improvements to include reductions in size, weight andoverall portability. Current technology should also provide a means ofmaking the equipment more rugged and maintenance free. These improvementsare essential in view of the overall burden placed on logistical support onthe modern battlefield.

APPROACH: Current engineering technology will be evaluated for applicationto the requirements of the dental cutting system which provides the drivingforce for the cutting instrument itself. Principles developed in fluidictechnology will be used as the basis for developing a more rugged main-tenance free system as well as reducing overall size and weight. Thesystem will be constructed by the Harry Diamond Laboratory.

RESULTS: A series of preliminary design criteria have been prepared whichwill be directed initially at elimination of much of the flexible tubing inthe current syste since this is its most damage prone area. Much of theremaining effort has been directed at consolidation and repackaging forstrength, reduced weight; and reduced cube. Construction of the prelimi-nary design will begin in FY83.

35

PROJECT: 3S162775ZA825

WORK UNIT TITLE: DEVELOPMENT AND EVAWIATION OF DENIAL MATERIEL FOR FIELDUSE

PRINCIPAL

INVESTIGATOR: MAJ MICHAEL P. RETHMAN, DC

Computerized Subtraction Radiography in the Diagnosis of

Bone Wounds in Dogs

PROBLEM: Studies have shown that enormous amounts of bony destruction areundetectable on conventionally viewed sequential radiographs. It has alsobeen shown that the effect of a camplex background of unchanged radiopaquematerial complicates the detection of subtle changes. Significant destruc-tion of hard tissues due to infection can often progress undetected withthe result that treatment time as well as lost duty time for the woundedsoldier are increased. A radiographic system which can provide earlydetection of bony destruction due to infection could be of considerablevalue in a field hospital. A technique has been developed whereby thedetection of subtle osseous changes is facilitated in sequential stan-dardized radiographs. Imagery remaining after this processing representsonly changed structures in this longitudinal effort. Investigations haveshown improved diagnostic capability in dry skulls using this methodology.The efficacy of this technique has not been demonstrated in a live modelnor does equipment or methodology exist to this end. The purpose of thisinvestigation is to develop necessary equipment and methodology to executea study comparing ccputer subtraction technique with conventional radio-graphic techniques in live models.

APPROACH: Highly standardized (geometric and densitcmetric) radiographsare a necessity for this study. Rigid interfacing devices are to bedesigned and built for use in the dog model. Methods of mathematical errorassessment are to be included. Osseous lesions (as wound anal'js) are tobe induced in dog mandibles. Preoperative and postoperative radiographsare to be compared as described in the PROBLEM4 statement. Statisticalanalysis of this comparison data will be generated. Analysis of theability of designed equipment and method to the end of highly standardizedradiographs will also be accomplished.

RESULTS: Preliminary results indicate that the equipment and methodologydeveloped for use in the dog will generate quality radiographs for use inthe study. Error assessment devices have been included in the equipmentand have demonstrated levels of geometric standardization in accuracyunapproached in the past. Densitametric standardization analyses arepresently underway. Two of the eight dogs are in the healing phase(postoperative) of the study. The remaining six dogs are scheduled forwound induction in November 1982.

36

f. AGENCY ACCESAICA I. OATE Of IUMNARV X NPOrT COaTrOL 5rMSOL

RESEARCH AND TECHNOLOGY W"RK UNIT SUMMARY DA OG 8672 82 10 01 DO-DR&E(AR)636L DATE PEMV 8wUY 0. KIND OF JMMAiV A IUMMAA ACTN WOU WINTY- O ASTRN 5. SPECIFIC ATA- LEVEL OF SUM

81 1001 D. CHMG~E JU U L N C.. .109UII0. *.COOCE* PROGRAM ELEU--T PROjECT MUN TASK AREA .M..CU WR UNIT MUNGE

IOMAN, 62775A 3_16-775A825 AB13I.u'..'.0 62775A 3S162775A825 AD

M/yyo I///II SIOG 80-7.2:5I1. TITLE (~.&* WO aN.a CIAUSANSNM C.W*

(U) Epidemiological Investigation of Dental EergenciesIS. SCIENTIFIC AND TECCINOS.OOICAL AMREAS*

012900 Physiology 012600 Pharmacologyi. START DATE 4. ESTIMATED COMPLETION DATE I. FUNING AGENCY 0IS. PE OIACe METHOD

81 10 I C T DA I C. IN-IfJSEI?. CONTRACT/IGANT 16. RESOURCES ESTIMATE M PAPOFESSIONAL MAN YRS L FUNDS (I. 0,..d.)

. SAYES/aFiICTIVI 611PINATtON* W......

M..MSC," FISCAL 82 1.0 40a TYP 4. AMOUNTi YEAQ

AEN, KILO AAR (.CUU. AMT. 83 1.2 75IS. RESPOMSLE 000 ORGANIZATION ML PEtPOMIG DRSiANI I

MA" USA Institute of Dental Research NAS'* USA Institute of Dental ResearchADS, Washington, DC 20012 ADDRESS Division of Clinical Operations

Washington, DC 20012PRINCIPAL INVSIAO 5~. SN91 .~ ue.-

NSOUN LE I DIVIAL "NA L: e

M SYweeey, T.P. TELEPSOM1! 202-576-3258TELEPONCI 202-576-3484 SOIAL SECURITY ACCOUNT WUNDER: C :DAII. EERNAL. tSE ASSOCIATE INVESTIGATORS

Foreign Intelligence Considered ..a, Carpenter, W.M.NAN, Grover, P.

axRYMMOS IF=(E-1ACE u- ...to 1I. (U) Lip Pathology; (U) Herpetic Lesions; (U) ColdWeather Survey; (U) Xerocheiliosis

N& TECHNICAL. OGECTIVE. 5 4S APPMOACH. IL 004i0611 (F.OR *"MEA I W0 8 lA . I rl0d 4 t .b F -*. Sl I. - Etl S-Io. ChM#"a.IeS CAIu .

23. (U) To determine the causes of dental emergencies in a population of soldiersreceiving regular dental care and to determine "at-risk-profiles" for those soldiersin critical occupation specialties so as to minimize problems with dental casualtiesduring deployment.

24. (U) Studies will be conducted amcng soldier populations to pinpoint the causes ofdental emergencies, their frequency and diagnostic strategies which will permit theprediction of the potential of each soldier for such emergencies. The goal is toselect out the "at-risk" group for dental treatment and thus minimize dental casualtiesduring deployment.

25. (U) (81 10 - 82 10) Cold weather surveys conducted at Ft Drum, New York were com-pleted in January 1982. A total of 1422 soldiers were surveyed. Preliminary analysisof data reveals there were 71% in the 20-29 age range, 98.5% were male and 78% spent50% or more time outdoors. The incidence of acute xerocheilosis was found to be 1.1%.The incidence of herpetic lip lesions was 3.1%. There is no marked difference inincidence rate when compared to complexion. Incidence rates increase with severity ofweather and increased exposure. The use of protective medication did not decrease theincidence of acute lip damage.

.SES.M. RI 0.US -atefw Nou *Wft s *mo,

D RAlam4 ANO 1l'441. I MAR 55 FOR ARMY USEI ARE OGSOLETE.

PRDJECT: 3S162775A825

WORK UNIT TITLE: EPIDEMOLOGICAL INVESTIGATIONS OF DENTAL EMERENCIES

PRINCIPALINVESTIGATOR: LTC DAVID M. LEWIS, DC

The Prevalence of Environmentally Induced Lip Pathology

Among Active Duty Soldiers: A Cold Weather Survey

PROBLEM: This study is necessary in order to establish base line datarelative to the prevalence of lip pathology amng the active duty militarypopulation. No data exist that identifies the prevalence of lip pathologyamong active duty service personnel and is related to occupational exposureto environmental factors. Military operations frequently require exposureof soldiers to extreme environment conditions for prolonged periods oftime. While the acute problems do not usually constitute a medicalemergency they do present problem with voice transnission, nutrition andmorale.

APPROACH: This is a survey of active duty personnel on field trainingexercises during adversely cold weather conditions. The age, sex, dutyenvironment, complexion, and lip condition will be recorded and weatherconditions correlated.

RESULTS: It appears that while actinic exposure is an etiologic factor inacute lip injury, the relative humidity of the environment is even moresignificant. Acute lip damage is prevalent among troops exposed to adverseenvironments. Apprecia6le amounts of acute damage were found in coldenvironments and hot dry climates. The data do not support the hypothesisthat sun exposure (actinic radiation) is the major risk factor. Relativehumidity appears to be a more significant factor based on both the hotweather and cold weather survey data. The modifying effects of caiplexionwere significant in cold weather, dark ccoplexion is a risk factor, whileit is a protective factor in hot weather. Age and amount of time spentoutdoors are not significant risk factors in acute cold weather lip injury.Based on these findings, the use of emollient preparations is recomended.Such preparations should protect the lips frcm acute lip damage caused bydessicaton in a dry climate regardless of the temperature.

38

I CY ACCESSION A OATE Of V napoar conO, sMoL,

RESEARCH AND TECHNOLOGY WORK UNIT SUMMARY DA OG 8674 82 10 01 DD-DR&E(AR)LJ68.ST RE UR . - -*OPWUMANY 8. S °MARY - -CNL TRACTOR ACCESS81 10 01 H. TERM OR QCUI

1W. NO./COOCBtS e'.RAN EL.EMENT o oj•c nUmoeN TAR AREA NVNUICK WORK UNIT NUMEER

0. P6A77YA 33162775A82 AA 012b. comuTG 62775A 3S162775A825 AB 012

STOG 80-7.2:_5,, ITE *N . - ,-.€-.---U--.ds~ (U) Development anc Evaluation of Meto and Materials ?or

KTreating Traumatic InjuryI& SCIENTIFIC AND TECHNOLOGICAL AREAS

010300 Miscellaneous Materials 002400 BioengineeringIL &TART DATE I4. ESTMAT U COMP.ETION OATE IS WuIA"O AIENCv IL- PE FOR -ACE NE OS

81 10 I82.09 DA I C. IN-lEUSEIf- C ACT ANT W. NSIOUNCES ISTIMATE PROFECSIONAL NAN VMS IL FUNOS (I. 6*4

A, OATC"FPECTIVC1 4EKPINATON:

I* "uUS R," FISCAL 82 1.0 61& TY P. a AMOUNT! Y A . V

ft EIN.F AWAM, f, CUM. AMT. 83 0 00. REWORUWE O00 OMMANI RATION . PC"FO NOOskmt 6 AN I I A I

"'ago USA Institute of Dental Research NAME" USA Institute of Dental ResearchWashington, DC 20012 Division of Oral Biology

Re.," ADOEMrI Washington, DC 20012

PRINCIPAL INVESYIGATOR (PWAf UANA 00 U.S. AAE. p..,.-V

MREPONS19LE INDIVIDUAL -a:* Tortorelli, A."me, Sweeney, T.P. TCELEPNOu: 202-576-3764.tELENOI, 202-576-3484 SOCIAL SECURITY ACCOUNT NUM.EM PC :DA

It. ENErMAL USE ASSOCIATE INVELTIOATONS

Fbreign Intelligence Considered %AMC, Vermilyea, S.

,L tTWOS,--ACW £LITV , *-150.. CON, (U) Polymer Devices; (U) Ceramic Devices;(U) Biodegradable Polymers; (U) Biodegradable Ceramic

EL TCNNICAL. OgJECTIVE. 1. APPROACN. IL PNO EM (rPkld.5 dW p. S wa- Ip 0 a*mA0 . p o *el# &4 A -J see' 5d*woft cl-* M C-

23. (U) To develop the technqlogy and methodolog for applying synthetic nmterials tothe effective management of maxillofacial injuries in the field.

24. (U) Biodegradable and partly biodegradable polymeric and ceramic devices will bedesigned and constructed. Designs will be directed at providing the potential forsimplifying the managnt of maxillofacial mounds from the point of initial injurythrough early definitive. treatment.

25. (U) (81 10 - 82 10) Testing was completed on a third generation device for thesegmental replacement of the trachea. This device was constructed of a polylactic-polyglycholic acid shell with polyvinyl chloride coated spring steel inserted in theshell to maintain patency. These prostheses as well as others of different design andconstruction which were previously reported have not been successful when implanted indogs. This particular effort has been terminated.. Additional devices planned fordesign and construction are being done umdercontract and will be evaluated in-house.These evaluations will be reported under DA Accession OH 6037 and OK 6020 in FY 83.

3904 :'2,14 8 ofo,, TooP I o 1,.o4om AR OoI.LaT 30 PORMS SA , Nov GoINAI MAI AMNO *LOI" 1 SiAN a0 AFOR ARMY USE) AMC OUSO LIETC


WORK UNIT TITLE: DEVELomET AND EVALUATION OF METHO[S AND MATERIALS FORTREATING TRAL1MATIC INJURY

PRINCIPALINVESTIGATOR: COL A. F. TORIORELLI, DC

Design and Construction of Prostheses

For the Reconstruction of the Trachea in Dogs

PROBLEM: Approximately 15% of all ccrnbat injuries are of the head andneck. A significant portion of these wounds will be avulsive. The devel-opment of materials and techniques to treat these wounds at the ccmbat sur-gical hospital level will result in a decrease in mortality and morbidityof ccmbat casualties. The trachea presents a particularly challengingproblem in devising a prosthesis for its replacement which will be inert,nonantigenic, allow no leaks, be rigid laterally but elastic anteroposteri-orly, have a noncollapsible lumen, and allow good fixation.

APPROACH: Several devices were constructed which had in camon a shell ofbiodegradable polylactic and/or polyglycolic acid. In order to provide forthe required rigidity, nondegradable frameworks were constructed of eitherceramic rings, a Teflon scaffolding, or a polyvinyl coated spring steelcoil. It was postulated that the biodegradable polymers would be slowlyreplaced by host tissue and inclose the nondegradable frame work.

RESULTS: Each prosthesis ised as its basic unit a cylinder constructed oflayers alternating between PLA polymer alone and mixtures of PLA-PGAcopolymer. The cylinders were prepared by spraying 10% solutions of thevarious polymer mixtures in methylene chloride onto a rotating mandrel ofappropriate dimensions. The resulting fibrous polymer cylinders were usedas inner cores slipped inside each of the three nondegradable system usedfor the different prostheses (ceramic rings, Teflon scaffold, polyvinylcoated spring steel). With the inner core in place inside the nondegrad-able system, a final outer core of polymer was placed on each device bycontinuing the polymer spraying procedure until the device had a wallthickness of 0.4am. The final prostheses were 5cm long with an insidediameter of 1.8cm. The ceramic rings used were constructed of alunina.The Teflon scaffold was constructed as a crib or partial cylinder in whichthe circumferential rings of the device did not meet. The spring steel wascomposed of medical grade stainless steel. All the devices were sterilizedby megadose gamma irradiation. The Teflon scaffolding was noticeablyermbrittled by the sterilization process. Ethylene oxide sterilization ofthe Teflon containing device caused separation of the Teflon from thepolymer. The polymer shells may have also been affected to same extent bythe gamma irradiation. All the devices constructed and subsequentlyimplanted in dogs failed within 6 weeks of placement.

40

0AGENCV ACCESAINO S. OATS o* SUNNA as.pp coamToL. suso,

RESEARCH AND TECHNOLOGY WORK UNIT SUMMARY DA OHI 6030 82 10 01 DD-DR&5(AR)*,S

5SAVE PREY SUWRY 4. KIND or gUmhARY L WNNARY SC? *OR TOCUR . NERA0NIN a BlWNMOWN 55 SPECI FIC OAT A-. LEVEL 01 .

81 100 D.U N(U U VL ONQ, A. "ItaUINT

14L NO./COOESe PROORAMN ELEMENT PROJECT NUmSaoR TASK AREA NUMISERI WoRK UNIT NUSeaR

b. COTOUII 6275 AD1277X8.- Efm"O ImSTOG 80_7 .2:5

1I. TITLE (f**w. "thSdESI

(U) Natural History of Oral Lesions Encountered in the SoldierIL. GCIETIC 44D TSCNNOLOGSCAL JIMIASN

012900 Physiology __________________

IL. START DATE JI.ESTIMATED COMPLETION OATE IL FUNUNGI AGENCY I.PERFORMANCE METHOD

690 ow DA 1 1 C. IN-HOUJSEIN. RESjORCEs eavuTIMA , PROFESSIONAL NAN VRS N& FUNDas (I . .

S DATEISAFEPCTIVEI EXPIRATIONR....

TYPE A ANOWNT, 'SAR

a KINGL P AWARDS I. CUMS. AMYT.83075

NAMEI USA Institute of Dental Research NAME,* USA Institute of Dental Research

SWashington, DC 20012 AMIRERE Washington, DC 20012

PRINCIPAL. INVEKSTIGATOR (Pr-WA SAJi It U.S. A-IC RIRDS.

.0 Carpenter, W.M.TELPAME

225638202-576-3484 ka. : 0257-38

TELEPHONE. 20-7-44SOCIAL SECURITY ACCOUNT NUSR: POC:DA1. 014HINA wagASSOCIATE INVESTIGATORS

Foreign Intelligence Considered NAME I Burnett, P.

ZXKLYOOS(-OtaAWW1%OM- -hJ~f -ft (U Ahous btw. 3. is; (U) Won ealig; (U eurrentAphthous Stcmtitis; (U) Lympho~cytes; (U) Laboratory Anima.l

SOL TECICAL OGJECTIVE6 IL APPROACH. JS. POGREM (FWN~d I.ER4WOAW VRIA hhNN00041 6, -. 0HSSR me# of 00..A I1fy Cs"086.N.. C d.)

23. (U) To recognize, characterize and develop effective therapeutic measures for thoslesions and conditions whicli affect the soldier due to military duty. The recognitionof environjmental and other factors which participate in the etiology of lesions andconditions unique to the military or are casually related to military duty will enablethe development of interceptive or therapeutic measures.

24. (U) To detect through clinical and/or microscopic observation oral lesions or acondition unique to the military population. To identify oral lesions or conditionswhich, though not unique to the soldier, are etiologically related to the performnceof duty. Onice identified the natural history including etiology, therapy, and prog-nosis will be established utilizing appropriate methods such as surveys, animal, andturmn investigations.

25. (U) (81 10 - 82 10) Idiopathic recurrent aphtbous stcmatitis (RAS) appears to be asignificantly greater problem in military as opposed to civilian populations and has anegative impact ca the healing of oral wunas. The involvmnt of immune ser.z and/orlymphocytes as possible mechanisms in the cytolyg4s of oral epethelial cells is beingevaluated by a radioisotope release assay using 'rcr. Results obtained to date sug-gest that blood nmnonuclear cells participate in the lvsis of autologous oral epithel-ial cells and is dependent on the presence of heat-stable factor(s) in the serumn ofRAS-prone individuals.

IA vNI S&SO to OWNS, dod" e mi. 4 1PRVOUS EDITIONSf OF THIS PORMA AREC OBSOCTE. 00 FORMS 1498A I NOY OfD D, "A8r14 98 AlSO 5500.0-1. 1 MAR Of IPOR ARM9Y USE I ARme OVSO NEYE.


WORK UNIT TITLE: NATURAL HISTORY OF ORAL LESIONS

PRINCIPALINVESTIGATOR: LTC PAUL R. BURNETT, DC

In Vitro Correlate of Recurrent Aphthous Stonatitis:

Evaluation of Immune Cytotoxicity by Radioisotope Release

PROBLEM: Studies show that the military loses approximately one millionman-hours per year to morbidity associated with maxillofacial injuries.Individuals prone to idiopathic recurrent oral ulceration or recurrentaphthous stomatitis (RAS), a group commonly believed to comprise a signi-ficantly greater proportion of the military than of the general population,manifest variably prolonged healing of oral wounds ranging from a few daysto several weeks. The objective of this research is to help reduce theimpact of oral and maxillofacial wounds on military manpower by elucidatingthe pathophysiological mechanism responsible for the amplification ofinflanmation, destruction of tissue, and acccopanying prolongation ofhealing associated with RAS.

APPRFOACH: A radioisotope ( 5 Cr)-release assay is being developed to quan-tifycytolysis within suspensions of nonkeratinizing oral epithelial cells.Using blood coaponents from paired RAS-prone and RAS-free subjects, thefour possible combinations of heat-inactivated serum with mononuclearleukocytes are assayed for their ability to lyse radiolabelled, autologousoral epithelial cells in order to evaluate the involvenent of immune serunand/or lymphocytes in this lytic process.

RESULTS: Results have been modest to this point, largely due to difficul-ties experienced in obtaining adequate quantities of isolated oral epithe-lial cells by trypsinization. The variability in cell yields has beenfound to correlate with the supplier-determined removal times for the vari-ous lots of trypsin we have used. Best results have been obtained withenzyme lots having recorded removal times of less than 10 minutes. Tryp-sinized oral epithelial cells incorporated SCr reasonably well; but thespontaneous release of the radioisotope has been somewhat greater thanoptimal. Preliminary data suggest that in our in vitro model, the partici-pation of blood mononuclear cells in the lysis0 autologous oral epithe-lial cells is dependent on the presence of a heat-stable factor(s) in theserum of RAS-prone individuals.

42

____ ~ ~ Gis.e., Accesse,OM 8OT T*tA~ *6PO~r corMOe PFMAO

RESEARCH AND TECHNOLOGY WORK UNIT SUMMARY DA OH 6037 82 106017 DD-DR&E(AR).ja

8110 01 D.CHANGE U U N' .... .c.. A. vat UI0s. "o./C00gel" PR*GRAM ELEMENT PftOjECT MNGMER TAMA AREA NUMBSIER Mone UNIT NUsmBee

bo~n PUA77Jbd(IA 007~5Ai2

S. C~..........

11711711, RiU 80-7.2:511 TTL ~-t afe-frcI.dBNsseo (d1 ,U ) New and Improve[ Techniques Lor uats and Boe Rgenera-

tion in 'fraizatic Wounds 3IL SCIESNTIPIC AND TECHOOGICAL AREAS"

012900 Physiology 002400 Bioengineer'n_______IL TAR? Aft 104. CITIMAT90 COMPLETION OATS IL. FUNIN A49ENCV 4 PcErfsIOASca WIETICO

69 01 'XtrD I I I *. NHOS-T. CONTRACT/BRMANT11.01UAISETMT "F61NL ANVS LFNDm

& OAEZtPCeVTEPIATIOII,...D

a tvPEc 4. AMOU*NT, VA

AKinclo ANARo. f.CUM. AMT. 8 . 1

M. seaSPOIUSLE DM 0OOBAMIZATION $8. PCAPOMNG @NB4ANIZATIO

Aes* USA Institute of Dental Research -*USA Institute of Dental Research

AMOB*Washington, DC 20012 Washington, DC 20012

PRINCIPAL 161VST IAT*R (FMb SIAN I# U.S. A &MC P1111AWS.J

REgPSPOOLt INRIVIGMAL "4;0 'Ibrtoeli , A.N-,Sweeney, T.P. TELECPHONE, 202-576-3764

aLleMI,202-576-3484 SOCIAL SCCUIT CCOUNT NUMER: PC:DA1. 4GENERAL WSE ASAOCIATC IMV9STI*ATOePJ

Foreign Intelligence Cosdee NANE, Woodyard, S.NAE0'Neal, R.

YORON(P(U BA- - ' icacium- Phosphate; (U) Ceramic Bl0ck; (U) Segment-al Mandibular Defects; (U) Granular Tricalcium Phosphate; (U) Laboratory Animals

83. TeCIONICAA oSECcTIvr." s. APPROACH. I&. PROBesmens (F"A ~.AM@*A 4p.,. § I FEO .AM. P-.4. As03 4 . 1 0 off -I&...se SC. C~o.)

23. (U) Current methodologies for anaging comnbat maxillofacial w~ounds and preventing/treating dental emergencies in thie field will be extrmly difficult to apply under theconditions anticipated in future war. New methods are required which will permit morerapid definitive care, reduce morbidity and decrease logistic load. Thus the objectiveof this wrk unit is to develop simrple, rapid methods for soft tissue or bone graftingutilizable by the dental specialist in the field.

24. (U) The fate, metabolism, osteogenic potenLal and tissue compatibility of ceramicand copolymer materials will be studied alone and in combination. The application ofthese and other mraterials to avulsive type wounds in both animals and humrans will bepursued.

25. (U) (81 10 - 82 10) Long-term evaluation (12-24 months) of biodegradable a-trical-ciiin phosphate (7CP) ceramic blocks for the segmental replacement of mandibular bonehas been comnpleted. Although biodegradation of and bone ingrowth into the ceramic isimproved by the presence of unidirectional porosi~ties in the ceramic, the process isvery slow. Approximately one to tw-thirds of the ceramic remins at 24 months post-placement. Healing is good but further inprovements in ceramic designs are needed andare being constructed. The use of granular 'ICP to treat bone defects around teeth hasbeen completed. It is concluded that with good post-operative care the defects can beresolved and tooth stability improved. Experiments are in progress to determine theefficacy of 7U' in stabilizing tooth implants in the presence and absence of traai.

PA OI -slaleA @-*-a* -P 66. , . 43

DD, 2^--41149 PNC.VI@UB 6OITIOMS OF 71415 FORM ARE OSS10CEYE00 P00RMB 14118A. I Nov as

I MAR AMO 400I.t. IMAR me ON0 hAllY USES ARE 03005 LT


WORK UNIT rITLE: NEW AND IMPROVED TECHNIQUES FOR GRAMT AND BONEREGENERATION IN TRAUMATIC WOUNDS

PRINCIPALINVESTIGATOR: COL A. F. TORTORELLI, DC

Tricalcium Phosphate Blocks in the Treatment of

Mandibular Discontinuities

PROBLE4: The loss of bone is a common sequelae of combat trauma. The useof autogenous bone where possible is a proven approach to resolving suchdefects. However, the extent of bone loss often makes this approachimpractical and the increased chance of morbidity due to the removal ofautogenous bone is a negative factor to be dealt with. The availability ofan off-the-shelf material which is plentiful, storage-stable, nontoxic andbiodegradable could provide a means of filling bone defects which wouldultimately be resolved as new bone. Tricalcium phosphate ceramic has beenstudied as a bone replacement material and found to be non-toxic and havean excellent shelf-life. However, it was found to biodegrade very slowlywith a tendency to crumble at a given point in time rather than maintainits integrity until ccimpletely replaced by bone. The block ceramic wasredesigned to include long unidirectional channels in the expectation thatthis would improve bone ingrowth and biodegradation properties in-vivo.

APPROACH: Ceramic blocks were used to replace surgically removed segmentsin the mandibles of dogs. The blocks were placed in the discontinuities insuch fashion as to align- the unidirectional channels (Approximately 300microns in diameter) parallel to the long axis of the mandible. Animalswere killed at intervals of 2,6,8,12 and 24 weeks and the tissues examinedhistologically for bone ingrowth. Long-term animals were maintained for 24months.

RESULTS: Previously reported results (Annual Report 1981) indicated cam-plete ingrowth of bone into the unidirectional channels at 24 weeks post-operative with only slight biodegradation of the ceramic. Renaining ani-mals have thus far been maintained for 24 months. Radiographic evaluationand sacrifice of one animal indicates that between 33 to 66% of the ceramicremains 24 months post-operative. However, the remaining material does notdemonstrate the catastrophic fragmentation previously noted using ceramicblocks without unidirectional porosity. While the results are prcmising,it is concluded that the slow biodegradation rate is not conducive to atimely and uneventful resolution of bone defects. Efforts are continuingto design a material with a more rapid degradation rate.

AA


WORK UNIT TITLE: NEW AND IMPROVED TECHNIQUES FOR GRAFTS AND BONEREGENERATION IN TRAUMATIC WOUNDS

PRINCIPALINVESTIGATOR: COL STEPHEN G. WOODYARD, DC

Biodegradable Ceramic Powder in Human Periodontal Defects

PROBLEM: The resolution of serious periodontal defects is possible onlywit- prolonged treatment. The accessibility of the periodontal defect tooral microorganism and the pocket configuration of the lesion site mili-tate against its ready resolution. The filling of these defects withautogenous bone has been demonstrated effective in resolving them. How-ever, the increased opportunity for morbidity provided by the harvesting ofautogenous bone make this approach questionable. A material which could insome way stimulate or precipitate osteogenesis and be available off-the-shelf could considerably reduce treatment time and provide a more effectivemnxoality for resolving periodontal defects.

APPROACH: Periodontal defects in a total of 51 individuals have beentreated with tricalciun phosphate ceramic in powdered form. Re-entry andbiopsy were taken at eight months post-operative and evaluated for osseousregeneration.

RESULTS: This study has been ongoing since 1974 and has been described inprevious reports. The last patients were scheduled for re-entry proceduresin March 1982. There were a number of difficulties. They included thefailure of some patients to return for final evaluation and the absence ofcontrols due to the necessity to treat all lesions observed rather thanallow them to progress. The overall results suggest that there was adecrease in pocket depth and increase in attachment levels in the samplepopulation. In no case were any undesirable effects noted. It is con-cluded that the tricalcium phosphate powder is useful in the resolution ofserious periodontal defects.

45

IAGENCY ^CC "G&174 O9 ATC OF *UNAR REPORT CorPOL .,.M,

RESEARC AND TECHNOLOGY WORK UNIT SUMMARY DA OK 6020 82 100 D-DR&E(AR)636

L*AsTEPREY SIWRY 4. 6150oPr R ARY a SUMMARY 6EUN 1EGAL $ W5INTR R PCIPIC OA A LIEVELI *F 6U,,

81 10 01 D. CHANGE NL ff ?CONO ACCESSUNI

40. N0./CoE5s' PROGRAM ELEMENT PRkOjECT MUSR TASKC AREA NUMOECR WORK UNIT NUMOUR_

"NSAfv 275 3S162775AS25 A ~b. CONTNISSJTIRG

ITITE (PR.ftE 4f R141 CIR.4IMMO c- d (U) Biodegradable Materials for the Treatment of Fractures and

IL. SCIENTIFIC ANW TECNNOLOOI4CAL AR A&

DAVESRPIPCTIVE, KRPIRATtON.sm

T YPEi A AMouNis YEa

&ENf&*PAWARD, I. CUk. Am T.83075

19. 01ESPONSISS.E boo 4"1GANAIAIOw 311 ICFQ . ..FM AIIAI-I

NANE.* USA Institute of Dental Research NRRIEF USA Institute of Dental ResearchDivision of Basic Sciences

AommCim. Washington, DC 20012 A00Rig' Washington, DC 20012PRINCIPAL INVIESIGAIOR (PCA& SSAOV It U.S. A£., MMI£*jv

M95PON06LLE INOIV40PAL NAMSR Tbrtorelli, A."a-L Sweeney, T.P. YELEPNONE: 202-576-3764TI[L9PN@NEI 202-576-3484 SOCIAL SE9CURITY ACCOUNT NUNSIER! POC :DA

f l I. GENERL USE ASSOCIATE INVIKSTIGATO S

Ebreign Intelligence Considered N.aE Hollinger, J.

AX KIUYONMS &A* 1 S- CI-II- '--(U) Biodegradable Copolymer; (U) Tracheal Grafts;(U) Diphospboinositide-Lsozyme: (U) Laboratory AnimalsZTECNICAL. OdJECTIVE.* 34 APPRO*ACN. SR. P010441921 (P 10 kl"11 0-1 I5 ~ . Sr .. G PF£. S- .. tl d -A 10R~I I.IRIEcE.)

23. (U) To develop rapid and ipp~roved methods of treating ccnbat injuries of the headand neck in the fieald using biodegradable materials. To develop premdicated biodegrad-able tissue fixation devices.

24. (U) Biodegradable polylactic acid, polyglycolic acid and various combinat ions ofthese polymers as well as other polymers being developed will be applied in the development of surgical procedures for a variety of hard tissue, soft tissue and hollow organinjuries in animals and extended to man where appropriate.

25. (U) (81 10 - 82 10) Studies on the segrwntal replacement of the trachea ir 10 dogshave not been successful in spite of a number of rmdifications of the device used.Most recently a combination of biodegradable copolymer (lactide-.co-glycolide) and coilespring steel were constructed to produce an open cylindrical device. This device wascalculated to be partially replaced by tissue ingrowth while retaining sufficient func-tional rigidity. All in vivo hollow organ studies have been terminated at this time.A study is in progress on the use of 50:50 poy-L(-)-lactide-c-o-glycolide containing adiphospboinositide-lysozyme cczalex for the segmntal replacement of ma~ndibular bone indogs. Necessary chemistries and presurgical preparations have been made and one imn-plant has been inserted.

D D, "NT 01498 PRE:VIOUS aDITIONS1 OPF TIS PoRM ARE OISOLETE. 00 PORM1R 1496A NOV 45AI54*&.1 ma MASO (won ARMYV USG) ARE OUS1OLETEK


WORK UNIT TITLE: BIODEGRAIABLE MATERIALS FOR THE TREATMENT OF FRACTURESAND SOFT TISSUE WOUNDS IN THE MILITARY SITUATION

PRINCIPALINVESTIGATOR: COL A. F. TORTORELLI, [C

Replacement of an Excised Tracheal Segment

in Dogs Using Combination Biodeqradable

and Biocompatible Prostheses

PRfBLEM: Hollow organ injuries such as segmental loss of the trachea aredifficult if not impossible to treat in some cases. A prosthetic replace-ment for the trachea would be a highly desirable means of treating seg-mental loss of the trachea resulting from ccmbat wounds. Many materialshave been used experimentally with varying degrees of success. To date nosatisfactory prostheses for the trachea has been developed. The use ofbiodegradable materials in a prosthetic device offers the possibility ofultimate replacement of the biamaterial with functional host tissue. Thepurpose of this research effort has been to determine whether a fibrillarnetwork of polylactic and polyglycolic acid copolymer inclosing a frameworkof biocanpatible material would allow for ingress and replacement by hosttissue with subsequent repair of the trachea.

APPROACH: Tracheal prostheses constructed with alternate layers of PLA andPLA-PA preformed into cylinders and placed within frameworks of alumina,Teflon or spring steel cbils; then sprayed with a polymer covering wereused to replace 3cm segments of the trachea in 10 dogs. The animals werefollowed postoperatively for functional capability until the prosthesesfailed. Histochernical evaluations were made at sacrifice.

RESULTS: Prosthesis using a framework of ceramic rings bound together by aPLA-PGA cylinder were placed in 3 animals. A Teflon frame formed into acrib like shape rather than a cylinder and enclosed within a PLA--_A coverwas placed in 3 animals. Finally a coil of spring steel was inclosed inPLA-PFA and placed in 4 animals. The animals survived an average of 4.3weeks with the longest survival at 5.6 weeks. In all cases advancingcyanosis and respiratory difficulties necessitated sacrifice. The pros-theses using ceramic rings failed for two reasons. As the polymer biode-graded it distorted, pulling the rings together and in same cases rotatingthem so as to occlude the lumen. Some, rings fractured and blocked thelumen. The Teflon cribs were apparently severely weakened by the method ofsterilization (gamma radiation) and crnbled within four weeks. The springsteel prosthesis were the most successful. However, polymer broke awayfrom the coils and occluded the lumen. Histological examination did revealtissue ingrowth into the polymer as early as two weeks postoperatively. Itcan be concluded that dimensional instability of the polymer cylinders dur-ing biodegradation was one of the principal causes of failure for all theprosthesis.

47

PRJECT: 3S162775A825

WORK UNIT TITLE: BIODEGRADABLE MATERIALS FOR THE TREATMENT OF FRACIURESAND SOFT TISSUE WOUNDS IN THE MILITARY SITUATION

PRINCIPALINVESTIGATOR: L I C JEFFREY 0. HOLLINGER, DC

A Study to Evaluate Copolymers of PLA-PGA and

Diphosphoinositide-Lysozyme Ccimplex for Bridging

a Surgically Prepared Bone Continuity Defect in the Mandible of Dogs

PROBLEM: Nearly 10% of all combat injuries are to the region of the headand neck. The mandible is frequently involved in these injuries.Reconstruction of discontinuity defects of the mandible is a challengingtask for military oral and maxillofacial surgeons. Restoration of thecontinuity of the mandible is paramount for prevention of scar contractureand to obviate mandibulofacial complex collapse. A variety of implantmaterials have been studied to determine their effectiveness for initiatingbone repair. Most of these materials have a significant drawback. Themost promising substance for bone induction appears to be a biocampatiblebiodegradable copolymer of PLA:PGA combined with an acidicphospholipid-lysozyme complex (DPI-L).

APPROACH: Seven female dogs under endotracheal nitrous oxide-oxygen andhalothane general anesthesia will have selected teeth extracted. Aftereight weeks, under similar anesthesia, a 2 an segment of the mandible withits periosteum will be surgically removed. An implant block of 50:50PLA:PGA-diphosphoinositide-lysozyme will then be fixed in the site using astainless steel plate and screws. Dogs will be sacrificed at 28, 42, 56,84, 120, and 168 days. At necropsy the implant site will be radiographedand clinically assessed and subsequently removed for histochemistry, totalprotein and hydroxyproline assay, enzyme analyses, cyclic nucleotideevaluation, and calcium to phosphate molar ratio will be determined.

RESULTS: To date one dog has been prepared for mandibular resection byextracting ipsilateral maxillary and mandibular dentition and performing analeveoloplasty. Eight weeks postoperatively a 2 cn segment of the mandiblewas removed and a PLA:PGA-DPI-L block was inserted in the defect. Reduc-tion and fixation have been accomplished and the animal is in function.The remaining animals will be prepared by the same protocol.

48

PRN AIIC, ACCESSI.0. j 0-C 0. S ... keON C.roT~ *VmL

RESEARCH AND TECHNOLOGY WORK UNIT SUMMAARY (n, PXI&1,,AH,6 6

S. OAYU wy 1U' 4. NINO ofs"G E aSa6 jI. f Act,- I 1 11 .0.0 IIcunty' I (606 RERDIG 041.. I6 ~ I(44 IN W. O"C

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A.PRMAY 6234A 3MI62734A875 Ab. CO TMISUTIAIG

k/56Y TOC FO-7-2-111. TITLE (P~- N1 Sfl7 c!***'11A C.&IIJ"

(U) Study of Saliva ai a Diaenostir Tol for Prp.'qpncp nf T.I-halAg1 n,VS. SCIENTIFIC AN0 TECHNOLOICAL. AREAS

... QQESTIMATEhDi y ~ COMPLETION DATE It '43lOIG AG(CC I If- -C1 ODA.CU11. STARnT DATEDA

791 7 CONT DC. IN-HOUSE7.CONt TRCTGRANT I* RESUR E AS -. 1111- -j(SOA IAM 1S - f

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66 NIOP *or .C . CUM. AMT. 83~.+Is_ 6i06Io~m,.g 008 ONG". l*?406 10 P4[RP0N610P061,ZAV"0

w-ogw*U.S. Army Institute of Dental Research AIAN-U.S. Army Institute of Dental Research

ADDRESS: Washington, DC 20307 ADDRESS* Washington. DC 20307

6(SPO.SISILt-I DA "ARE4.* * Wynkoop, J.R.--- SWEENEY, T.P. VfLE-. (202) 576-3393

(202) 576-3484 SOCIAL SE11T ACCOV436'6m*s4 pOjCDA21 Cl~qiIRA . as ASSOCI.hI "iS1.GA '0-$

A1MILLER, R.A.FIG .. SETTERSTROM- I A_

fono O$(p..C. ZAC #&S-0'r CI ... In--C046 (U) Salivary Amylase; (U) Nerve Agent; (U) SalivaryPhysiology; (U) Salivary Enzyme; (U) Laboratory Animal

21. CCMSICA.. OBJECTIVE.* 24 APPMOAC. SS P006.ess (F.- : h* N*fllA t4* INt - 1 C,. ...*. . * CI*~4t~

23. (U) To determine if saliva can be used as a diagnostic tool in evaluating theexposure of combat troops to chemical agents. To determine if constituents insaliva can be used to monitor the progress of therapy for chemical agent exposure.Develop a rapid, simplified field technique for diagnosis of chemical agent exposurein the combat soldier.

24. (U) Changes in saliva produced by chemical agents and prophylactic antidoteswill be evaluated. The particular areas of study will be enzyme, nucleotide, andprotein components. Possible methodology developed will be evaluated in the fieldand at the hospital level.

25. (U) (81 10 - 82 10) Work to date has suggested that variations in Salivaryamylase/total protein ratios may serve as the basis for a method for the diagnosisof chemical agent exposure in the field. Basal levels of several enzyme systemshave been evaluated in the saliva of anesthetized Rhesus monkeys. In addition,these levels have been compared to experimental levels of the same systems followingexposure to one.-of the chemical agents. Work is in progress to determine theeffects of prophylactic pretreatment upon the salivary enzyme systems previouslyexamined. Future work will compare enzyme levels from the current pretreatmentstudy with levels from animals pretreated and subsequently exposed to a chemicalagent.

PA -. "-'-- -1- -- - 149

DD, 2'.A41 498 A..,c.sr~0 ,'.. . .

PROJECT: 3M162734A875

WORK UNIT TITLE: A STUDY OF SALIVA AS A DIAGNOSTIC TOOL FOR THE PRESENCEOF LETHAL AGENTS

PRINCIPALINVESTIGATOR: CPT JUDSON R. WYNKOOP, DC

A Study of Saliva as a Diagnostic Tool for the

Presence of Lethal Agents

PROBLEM: The possible use of chemical warfare agents is a very real andpotentially devastating aspect of modern warfare. The current method ofdiagnosing chemical agent intoxication is by clinical signs of anticholin-esterase poisoning including increased secretions and muscle fascicula-tions. However, without specific prior knowledge of patient contact withchemical agents, diagnosis is tenuous. The situation is further compli-cated by the fact that certain prophylactic chemicals elicit signs andsymptoms similar to the agents. Both situations are unacceptable. If a"dip stick" could be developed that would rapidly test salivary secretionsfor the effects of chemical agents, then the confusion surrounding a diag-nosis of poisoning would not occur.

APPROACH: In an attempt to develop a fast, accurate diagnostic test forchemical agent poisoning, specific biochemical constituents in Rhesussaliva have been studied before and after agent intoxication. Theseconstituents were found in parotid and extraparotid saliva specimens whichwere collected during anesthesia base-lines and incremental dosings ofspecific agent. Flow rates were measured at each of the samplings and thefollowing tests were done on the specimens: (1) total protein, (2) amylaseactivity, (3) amylase zymogram, (4) lysozyme activity, (5) kallikreinactivity, and (6) cyclic AMP.

RESULTS: Coaparing base-line values of rhesus saliva collected under twowidel -used anesthetics, it is evident that each anesthetic causes a dis-tinct secretion frcm the parotid glands as well as the other major salivaryglands. The parotid and extraparotid flow rates for the Ketalar injectedanimals were significantly higher (p-.006 and p-.0001, respectively) thanthe flow rates of the Innovar injected animals. In general, the total pro-tein and enzyme activities of the salivas collected from Innovar injectedanimals had higher values than those injected with Ketalar. The resultsindicate that Innovar slows the flow rate and indirectly increases theenzyme activity in the salivas. Ccmparing the values obtained from theKetalar :njected animals with the values obtained from the saliva ofanimals injected with a specific agent, it should be noted that the flowrates of the agent treated animals decreased consistently fran the base-line values. The amylase/protein ratios in parotid saliva demonstrated asignificant decrease with agent.

53

PUBLICATIONS

SUBMITTED:

1. Grover, P.S., Carpenter, W.M., and Allen, G.W.: PanographicSurvey of U. S. Army Recruits: Analysis of Dental Health Status.Milit Med

2. Grover, P.S., Carpenter, W.M., and Allen, G.W.: DentalEmergencies Occurring Among U. S. Army Recruits. Milit Med

3. Grover, P.S., Carpenter, W.M., and Allen, G.W.: Predictability ofDental Emergencies by Panography. J Dent Res

4. Hollinger, J.O. and Gee, S.A.: Evaluation of the Parietal Bonesin the Rat as a Specific Site for the Testing of OsteogenicMaterials SHORT TITLE: A Simple Animal Model to Study BoneImplant Material. J Dent Res

5. Hollinger, J.O.: In Vivo Calcification Induced by a ProteolipidCcoplex (Lysozyme-Acidic Phospholipid). Bicmater, Mea Devices,Artif Organs

6. Hollinger, J.O.: A Preliminary Report on the Osteogenic Potentialof a Biodegradable Copolymer of Polylactide:Polyglycolide(PLA:PGA). J Bicmed Mater Res

7. Mader, C.: Mandibular Talon Cusp. J Am Dent Assoc

8. Finley, S.L. and Hollinger, J.O.: Separation of AlkalinePhosphatase Isoenzymes from Various Rat Tissues Using Flat-BedAcrylamide Gel Isoelectric Focusing. J Dent Res

9. Russell, M., Lorton, L., St. Hoyme, L.E., and Snyder, A.J.: TheCorrelation Between Bone Loss and Tooth Malalignment. J PeriodontRes

10. Vermilyea, S.G., Kuffler, M.J., and Tamura, J.J.: Observations of"Economical" Fixed Prosthodontic Alloys. Milit Med

11. Lorton, L., Peters, D.D., and Wong, M.: Testing the Sturdiness ofClinical Criteria with Different Methods of Simple StatisticalAnalysis. J Dent Educ

12. Grover, P.S. and Lorton, L.: A Bifid Mandibular Nerve as aPossible Cause of Anesthetic Deficiency in the Mandible. J OralSurg

13. Stalkler, W.H. and Allen, G.W.: Bilateral Blind EpithelializedTracts Associated with the Inferior Labial Frenum: Report of aCase. Oral Surg

451

14. Lorton, L. and DeNucci, D.: Pre-Cementation Stabilizing ofEtch-On Fixed Prostheses. J Prosth Dent

15. Peters, D.D., Lorton, L., Mader, C.L., Augsburger, R.A., andIngram, T.A.: Evaluation of the Effects of Utilizing CarbonDioxide as a Pulpal Test. Part I: In Vitro Effect on HumanEnamel Surface. J Endodont

16. Ingram, T. and Peters, D.D.: Evaluation of the Effects ofUtilizing Carbon Dioxide as a Pulpal Test. Part II: In VivoEffect on Canine Enamel and Pulpal Tissues. J Endodont

17. Grover, P.S. and Lorton, L.: Impaction of First Permanent Molars.A Case Report. Oral Surg

18. Shulman, J.D.: Linear Progranming: A Resource AllocationMethodology for Dental Managers. J Pub Health Dent

19. Grover, P.S., Lorton, L., and Hollinger, J.: Incidence of Pain orDiscomfort Following One-Visit Operative Treatment: A Review. JProsthet Dent

20. Grover, P.S., Lorton, L., and Hollinger, J.: Incidence of Pain orDiscomfort Following One-Visit Operative Treatment: A ClinicalStudy. J Prosthet Dent

21. Vincent, Jack W., Falkler, W.A. Jr., and Craig, J.A.: Comparisonof Serological Reactions of Typed Fusobacterium nucleatun Strainswith Those of Isolates from Humans, Canines, and a Macaca mulattaMonkey. J Clin Microbiol

IN PRESS

1. DiFiore, P.MN., Peters, D.D., Setterstrom, J.A., and Lorton, L.:The Antibacterial Effects of Calcium Hydroxide ApexificationPastes on Streptococcus sanguis. Oral Surg

2. Vermilyea, S.G., Kuffler, M.J, and Tamura, J.J.: Techniques forthe Accurate Casting of Base-Metal Alloys. Milit Med

3. Guinn, J.W.III, Griswold, W.H., and Vermilyea, S.G.: The Effectof Cooling Rate on the Apparent Bond Strength of Porcelain-MetalCouples. J Prosthet Dent

4. Vermilyea, S.G., Tamura, J.J., and Mills, D.E.: Observations onNickel-Free Beryllium-Free Alloys for Fixed Prostheses. J Am DentAssoc

52

ARTICLES

1. Freccia, W.F., Peters, D.D., and Lorton, L.: An Evaluation ofVarious Permanent Restorative Materials' Effect on the Shade ofBleached Teeth. J Endodont 8:265-268, 1982.

2. Machian G.R., Peters, D.D., and Lorton, L.: The ComparativeEfficiency of Four Types of Endodontic Instruments. J Endodont8:398-402, 1982.

3. Setterstram, J.A.: Controlled Release of Antibiotics fromBiodegradable Microcapsules for Wound Infection Control. ArmyScience Conference Proceedings, 111:215. 1982.

4. Skirvin, D.R., Vermilyea, S.G., and Brady, R.E.: Polymethyl-methacrylate Reinforcement; Effect on Fatigue Failure. Milit Med142: 1037-1040, 1982.

ABSTRACTS

PRESENTED AT THE ANNUAL MEETING OF THEAMERICAN ASSOCIATION FOR DENTAL RESEARCH

PUBLISHED IN THE JOURNAL OF DENTAL RESEARCH, SPECIAL ISSUE 61, 1982

1. Tortorelli, A.F.: Biodegradable Materials Research in the U. S.Army - A Review.

2. Setterstrcm, J.A.: In Vivo Evaluation of Controlled ReleaseMicroencapsulated Antibiotics Topically Applied to Wounds.

3. Hawley, C.E.: Factors in Bone Growth and Repair: The Challengefor the Military in the Future.

4. Finley, S.L.* and Hollinger, J.O.: Separation of AlkalinePhosphatase of Rat Tissues by Isoelectric Focusing (IEF).

5. Hollinger, J.O.: The Osteogenic Potential of a 50:50 PolylacticAcid-Polyglycolic Acid (PLA:PGA) and Diphosphoinositide-LysozymeComplex.

6. Hawley, C.E. and Heath, J.R.*: Isolation of Active OsteoclastActivating Factor (OAF) Subunits by Ultrafiltration.

I

7. Miller, R.A.*, Wynkoop, J.R. II, and Johannsen, K.: Analysis ofHydroxyproline in Gingival Exudate by High Performance LiquidChromatography.

8. Wynkoop, J.R.II*, Miller, R.A., Cheong, V., and Lorton, L.:Levels of Neuroactive Substances Following Exposure to MethylMethacrylate Monomer.

53

4L mm mm

9. Wynkoop, J.R.II, Woodyard, S.*, and Miller, R.A.: A Ccuparison ofIndividual Crevicular Fluid Hycroxyproline Levels to PeriodontalAssessments.

10. Miller, R.A., Wynkoop, J.R.II, Allen, G.A.*, and Lorton, L.: SomeEffects of Methyl Methacrylate Monomer Upon Renal Function.

11. Tortorelli, A.F.* and-Grower, M.F.: Attempts at Replacement of anExcised Tracheal Segment in Dogs Using Combination Biodegradableand Biocampatible Prostheses. Proceedings of 14th InternationalBicmaterials Symposium, Society for Bicmaterials.

12. Grover, P.S.* and Hollinger, J.O.: The Potential of a ProteolipidCcmplex (Lysozyme-Acidic Phospholipid) to Produce Osteogenesis.Proceedings of 14th International Bianaterials Symposium, Societyfor Bicniaterials.

13. Hollinger, J.O.: The Osteogenic Potential of a Ccuplex Consistingof a Biodegradable Copolymer and Diphosphoinositide-Lysozyme.Proceedings of 14th International Biamaterials Symposium, Societyfor Bicmaterials.

54

• --- -----, . -- m ,M mm, m , mnmmmm 4L m -I- - ,-- nn n nu

DISTRIBUTION LIST

5 copies to:

US Army Medical Research and Developnent ConandFort Detrick, Frederick, MD 21701

12 copies to:

Defense Technical Information CenterATIN: DDC-DDAAlexandria, VA 22314

1 copy to:

SuperintendentAcademy of Health Sciences, US ArmyAT1N: AHS-COMFort Sam Houston, TX 78234

1 copy to:

DirectorBiological & Medical Sciences DivisionOffice of Naval Research800 N. Quincy StreetArlington, VA 22217

1 copy to:

CommanderNaval Medical R&D CommandNational Naval Medical CenterBethesda, MD 20014

1 copy to:

HQ AFMSC/SGPABrooks AFB, TX 78235

i copy to:

Director of Defense Research and EngineeringATqN: Asst Dirctor (Environmental and Life Sciences)Washington, DC 20301

55

i, l eiiiadz - dtic · the number of osteoclasts and heightens their ltic activity. ccnbat injury...

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