nanotechnology in biomedical research

7/24/2019 Nanotechnology in Biomedical Research

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Nanotechnology in BioMedical SciencesNanotechnology in BioMedical Sciences

Earl J. Bergey, Bruce A. Holm, Tapas De, Yudhisthira Sahoo, DavidRodman,Christopher Friend, Indrajit Roy, and Paras N. Prasad

Institute for Lasers, Photonics and Biophotonics and

School of Medicine and Biomedical SciencesThe State University of New York at Buffalo

Buffalo, New York 14260


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AbstractAbstract

Nanotechnology is rapidly expanding into the biomedical field. At the Institute for Lasers,Photonics and Biophotonics, research in nanotechnology has focused on the development ofmultifunctional nano-sized particles we have termed as nanoclinics. They consist of a nanosizedbubble fabricated to carry therapeutic or diagnostic components to targeted cells. The outer surfaceof these particles can be functionalized to incorporate biologically active targeting agent. Thistechnology provides a platform for the development of new imaging, diagnostic and therapeuticmodalities. For bioimaging and diagnostics, nanoclinics are fabricated containing rare-earth ions,which exhibit two-photon, anti-stokes luminescence by frequency up-converting infrared to visiblelight. At the Institute, we have successfully prepared the phosphors containing nanoclinics having asize ~25nm with a silica shell around it which helps in aqueous dispersability and also allows

functionalization for covalent attachment of bioprobes for targeting. The coupling of specificpeptides, proteins or nucleic acid sequences to the silica shell will allow for the selective detectionof biological entities that will have applications in microscopy, flow cytometry, ELISA andDNA/RNA hybridization systems. This adaptable nanoparticle platform was used to developnanoclinics, which selectively target specific cell or tissue types. These targeted nanoclinics cancarry therapeutic drugs, DNA for gene transfer therapy, or provide externally activated therapy as inphotodynamic therapy or magnetic field activation. The core of our prototypic nanoclinic consists

of either Fe2O3 or Fe3O4 to which a fluorophore is coupled. These nanoparticles are encased ineither silica or by a fatty acid bilayer, respectively. The final step is to attach the targeting ligandLH-RH (luteinizing hormone-releasing hormone) to complete the nanoclinic. This design allowsfor detection of specific cells or tissues using standard MRI equipment and can be usedtherapeutically with the same instrumentation (stronger DC magnetic field). These nanoclinicscause magnetocytolysis of LH-RH receptor-positive cancers, in vitro, in a DC magnetic field..These therapeutic nanoclinic systems could provide the clinician ith a ne tool in the fight againsthuman diseases.


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NanoClinicsNanoClinics

Composed of !ilica or Ca"#$shell encapsulating , Rare-earth

elements, iron o%ide, D&' or DrugsLess than ( nm in size *tuna+le)

"roduced +y a reverse micelle process

!urface can +e modified *targeting)

Rare arth elements fficient upconversion *+lue, red and green)

!ta+le *little photo+leaching)

Diagnostic 'pplications in ioimaging and !ensing in/icroscopy, 0lo Cytometry, L1!', 0luorescent 1magingsystems, /R1 contrast agent

Therapeutic 'pplications in 2ene therapy, Drug Delivery!ystems, "hotodynamic Therapy, and Direct Cancer Therapy


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First Generation of NanoclinicsFirst Generation of Nanoclinics

Cancer diagnosis and targeted therapy


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Luteinizing Hormone!eleasing Hormone "LH!H#Luteinizing Hormone!eleasing Hormone "LH!H#

$argeted Nanoclinic$argeted Nanoclinic

!ilica

LHRH

&anoclinic

To photon dye

1ron #%ide

LHRH

LHRH


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%pta&e of LH!HNanoclinics in MCF' Cells%pta&e of LH!HNanoclinics in MCF' Cells

3min 27min21min15min9min

"articles saturation ith time

T"L!/ image of

+reast carcinoma cells.

The arro shos

nanoparticles

accumulating on and

inside the cell.

transmission Fluorescence


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0

1

2

3

4

5

6

0

1

2

3

4

5

6

-

+

+

LH-RH

LH-RH

LH-RH

Num

ber

ofCe

lls

Lysed

(10

4!

"2

#rs$

2

#rs$

%C

&-10"

'C-"

'

C-"

%C

&-10"

Num

ber

ofCe

lls

Lysed

(10

3!

Specificity of Magnetocytolysis by NanoclinicsSpecificity of Magnetocytolysis by Nanoclinics

Nanoclinics incubated (ith adherent cell for )* hours+ Cells (ashed, released by

trypsinization and sub-ected to .C magnetic field+ Magnetocytolysis determined by

counting remaining cells+

MCF'/ Human breast carcinoma LH!H receptor positi0e%C123'/ Human uterine carcinoma-LH!H receptor negati0e


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0

2

4

6

)

10

12 -*C LH-RH + LH-RH

Num

bero

fLyse

dCe

lls

(10

3!

4ffect of %p and .o(n !egulation of the LH!H !eceptors4ffect of %p and .o(n !egulation of the LH!H !eceptors

on Magnetocytolytic Acti0ity of LH!HNanoclinicson Magnetocytolytic Acti0ity of LH!HNanoclinics

4GF "epidermal gro(th factor# 5 upregulation

!C263 "synthetic somatostatin# 5 do(nregulation


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Surface $opography of MCF' Cells 1ncubated (ithSurface $opography of MCF' Cells 1ncubated (ith

LH!HNanoclinics AFMLH!HNanoclinics AFM


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/*au)

H*2auss)

.3

.$

.4

.5

6

6.3

( 6 6( 3 3(

a

a

ab

b

"articles ith cells

"articles in media

b

a +

Free particlesCell Bound particles

cell cell

4ffect of Cellular 1nteraction4ffect of Cellular 1nteraction

Magnetic 7roperties of NanoclinicsMagnetic 7roperties of Nanoclinics


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Tm78+983#:

r983#:r78+983#:

%pcon0ersion 4mission by%pcon0ersion 4mission by

!areearth Nanophosphors!areearth Nanophosphors

Tm - thulium

r - er+ium

8+ - ytter+ium

83#:- yttrium o%ide


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%con,erin. Nanoec#nolo.y for%con,erin. Nanoec#nolo.y for

/io-&ma.in./io-&ma.in.

; Cellsmission from

&anophosphor

; Cells and

mission from

&anophosphor


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$4M of .NA.oped Ca78$4M of .NA.oped Ca78**NanoparticlesNanoparticles


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In VitroIn Vitro$ransfection 4fficiency of$ransfection 4fficiency of

.NA.oped Ca78.NA.oped Ca78** NanoparticlesNanoparticles

Cell line 9 :ur&at

.NA 5 pS;

gal "

galactosidase#7ositi0e Control 5 Superfect See acknowledgementsSee acknowledgements


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In VivoIn Vivo$ransfection (ith .NA.oped Ca78$ransfection (ith .NA.oped Ca78**Nanoparticles andNanoparticles and

GalactoseSurface Modified .NA.oped Ca78GalactoseSurface Modified .NA.oped Ca78**NanoparticlesNanoparticles

!ee ac


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ConclusionsConclusions

Nanoclinics pro0ide the follo(ing ad0antages/

/ultifunctional nanoparticulate platform

!urface modifia+le for coupling of targeting agents Can carry different payloads *Therapeutic or 1maging)

Can e%ert therapy using e%ternal activation

>p-conversion +ioimaging ith reduced +ac


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Ac&no(ledgementsAc&no(ledgements

$he gene transfer studies (ere performed by .r+ 1 !oyunder the direction of /

Dr. '.&. /aitra

Department of Chemistry

>niversity of Delhi

Delhi, 1ndia

as partial re

nanotechnology in biomedical research

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