56S Proceedings of the NASS 22nd Annual Meeting / The Spine Journal 7 (2007) 1S–163S

METHODS: The volunteers filled out our original questionnaire and ex-

amined neurological findings of BTR, TTR, PTR, ATR, Hoffmann and

Wertenberg reflex. The questionnaire consisted of following 2 questions;

(1) ‘‘do you have any pain or numbness in your neck, shoulder, arm or

hand in a last week?’’, (2) ‘‘do you have any difficulties with buttoning

your dress, using a fork or spoon, walking, one foot standing or stairway

gait?’’. The volunteers who answered ‘‘yes’’ both above questions and

had one of positive neurological findings were defined as ‘‘myelopathy

suspect’’. Statistical significance level was set at 0.05 for chi-square test.

RESULTS: The rate of respondents ‘‘yes’’ was 6.0% under forty, 7.3% in

forties, 13% in fifties, 16% in sixties, 30% in seventies and eighties. Statis-

tical significance was found. The rate of the volunteers with positive neuro-

logical findings was 31% under forty, 25% in forties, 32% in fifties, 27% in

sixties, 19% in seventies and eighties. The rate of ‘‘myelopathy suspects’’

was 3.6% under forty, 1.6% in forties, 4.2% in fifties, 3.8% in sixties,

5.0% in seventies and eighties. Statistical significance was not found.

CONCLUSIONS: Morbidity of symptoms suspected of cervical myelop-

athy did not increase with aging in the countryside of Japan.

FDA DEVICE/DRUG STATUS: This abstract does not discuss or include

any applicable devices or drugs.

doi: 10.1016/j.spinee.2007.07.135

Friday, October 26, 200710:25–11:05 AM

General Session: Biologics - Disc Regenerationand Repair

115. Nucleus Pulposus Regeneration Using Fresh Autologous

Adipose-derived Cells

Hans-Jorg Meisel, MD, PhD1, Tim Ganey2, William Hutton3,

Tim Moseley4, Marc Hedrick4; 1Berufsgenossenschaftliche Kliniken

Bergmannstrost, Halle, Germany; 2Atlanta Medical Center, Atlanta, GA,

USA; 3Atlanta Veterans Affairs Hospital, Decatur, GA, USA; 4Cytori

Therapeutics, San Diego, CA, USA

PURPOSE: Adipose-derived stem and regenerative cells (ADRCs) differ-

entiate into a nucleus pulposus-like phenotype when exposed to in vivo en-

vironmental factors. A study was initiated to evaluate whether autologous

cells derived from adipose tissue would be effective in an established dog

model for acute disc injury.

METHODS: Adipose tissue was harvested in 4 adult dogs (25–30 kg)

from the thigh and ADRCs were collected from the tissue using enzymatic

digestion and cell concentration. Nucleotomy was performed on three

levels. One million cells were suspended in either (a) hyaluronic acid

(HA) or (b) saline, and placed centrally into two of the three levels.

MRI was performed on all four dogs at six and twelve weeks. Dogs were

euthanized at six and twelve weeks following the placement of the cells.

Radiographic anatomy, MRI, histology and gross pathology were used to

assess the outcomes of the intervention.

RESULTS: Disc levels receiving cells with HA carrier had thicker discs,

a higher cell content with better organized colonies of cells, more matrix pro-

duction compared to the levels receiving cells with a saline carrier. Levels

that had only the nucleotomy lacked significant regeneration of matrix.

CONCLUSIONS: This feasibility study provides additional evidence that

ADRCs can be transplanted at the time of surgery and injected safely by

percutaneous means. Coupled with encouraging results described here,

plus other clinical studies using autologous disc chondrocyte transplanta-

tion, this project has now been extended to include six-month and one-year

assessments of ADRCs injected with an HA carrier.

FDA DEVICE/DRUG STATUS: This abstract does not discuss or include

any applicable devices or drugs.

doi: 10.1016/j.spinee.2007.07.137

116. The Action of Resveratrol, A Phytoestrogen Found in Grapes

and Red Wine, on the Intervertebral Disc

Frank Phillips, MD1, Hyun-Hee Kim2, Xin Lee2, Howard An, MD3,

Hee-Jeong Im Sampen2; 1Rush University Medical Center, Chicago, IL,

USA; 2Chicago,, IL, USA; 3Chicago, IL, USA

BACKGROUND CONTEXT: Degeneration of the intervertebral disc

(IVD) is considered a competent cause of back pain. With disc degenera-

tion, anabolic processes are inhibited and catabolic factors such as metal-

loproteases (MMPs) are up regulated. These result in progressive loss of

proteoglycan (PG). The phytoestrogen resveratrol (trans-3,40,5-trihydrox-

ystilbene) is a natural polyphenol compound found in various plants in-

cluding grapes. It’s anti-inflammatory, anti-catabolic and antioxidant

properties have been documented and it is believed to be one of the com-

pounds that explain the health benefits of moderate red wine consumption.

PURPOSE: The purpose of the study is to determine the effect of resver-

atrol on IVD regeneration by assessing the production of PGs and MMP-13

and also evaluate whether resveratrol counteracts catabolic factors such as

basic fibroblast growth factor (bFGF) or IL-1 in bovine IVD chondrocytes.

STUDY DESIGN/SETTING: A biologic study assessing the effects of

resveratrol on anabolic and catabolic pathways in the IVD.

METHODS: IVD tissue was harvested from bovine coccygeal tissue and

chondrocytes were isolated from nucleus pulposus (NP). After enzymatic

digestion, cells were resuspended in 1.2% low viscosity sterile alginate

beads at 2 million cells/ml. The alginate beads were cultured in complete

medium with media changes at two day intervals. After 5 days of pre-cul-

ture, the beads (quadruplicate culture, 10 beads each) were treated with

different growth factors and cytokines in the presence or absence of resver-

atrol in conditioned medium containing 1% FBS for 14 days. BMP7 was

used as a reference. PG content was measured with Dimethylmethylene

Blue assay and normalized to DNA content. DNA assay representing cel-

lular proliferation was performed using PicoGreen (Molecular Probes).

Cell survival was measured using Calcein AM to stain live cells and ethid-

ium bromide homodimer 1 to stain dead cells. Anti-MMP-13 antibody

(R&D System) immunoblotting was performed. ANOVA was performed

using StatView 5.0 software (SAS Institute, NC). P values less than 0.05

were considered significant.

RESULTS: Incubation of NP cells with resveratrol for 14 days signif-

icantly promoted PG production in a dose dependent manner. PG pro-

duction in cells treated with resveratrol at a concentration of 100uM

was greater than in cells incubated with BMP7 (100 ng/ml), suggesting

a potent anabolic impact on NP cells. There was no effect of resveratrol

on cellular proliferation. Incubation with resveratrol greatly promoted

cell survival. Stimulation of NP cells with bFGF or IL-1 augmented

the production of MMP-13. This augmentation of MMP-13 was signif-

icantly reduced by resveratrol in a dose-dependent manner. Exclusion

assays that visualize the formation of cell-associated matrix further con-

firmed the anabolic effect of resveratrol and its antagonistic action on

bFGF and IL-1.

CONCLUSIONS: Resveratrol which is found in grapes and red wine was

found to be a potent stimulator of PG production and improve cell viability

in bovine NP cells cultured in alginate beads. In addition resveratol was

able to reverse the catabolic effects of bFGF and IL-1 that have been im-

plicated in disc degeneration. Resveratrol imbibed in red wine has been

proposed to have beneficial effects on various organ systems and warrants

further study for its effect on the IVD.

FDA DEVICE/DRUG STATUS: This abstract does not discuss or include

any applicable devices or drugs.

doi: 10.1016/j.spinee.2007.07.138

117. EuroDisc study – Assessment of Efficacy and Safety of

Sequestrectomy Plus Autologous Disc Chondrocytes – Second

Analysis of a Subgroup

Rudolf Bertagnoli, MD, PhD1, Hans-Joerg Meisel, MD, PhD2,

H. Michael Mayer3, Axel Reinhardt, MD4, Friedrich Weber, MD5,