Nanomedicine
A vision for future health, utilizing cross-fertilization of
nanotechnology and biology to produce novel
approaches
• for probing biological processes at the
molecular, subcellular and cellular levels.
• For sensing and bioimaging of biological events
• For incorporating multimodal diagnostics
• For implementing effective and safe targeted
gene therapy
NANOPHOTONICS AND NANOMEDICINE
Control of Optical Transitions
• Quantum Dots for Bioimaging
Novel Optical Resources
• Rare-earth up-converters for bioimaging
• Plasmonic nanoparticles for biosensing or therapy
Nanocontrol of Excitation Dynamics
• Nanoscopic sub-cellular interactions using FRET
• Nonlinear optical techniques for bioimaging and light-activated therapy
Manipulation of Light Propagation
• Biosensing using photonic crystals
Nanoscopic Field Enhancement
• Plasmonic enhancement for apertureless near field
• Plasmonic enhancement for Raman and fluorescence
Multimodal Imaging
COOH
OH
HOOC
HOOC OH
COOH
HOOC COOH
HOOC
OH
HOOC
COOH
HOOC
HO
COOH
COOH
HOOC
HO
COOH
HOOC
i
Fluorescent dye
Fe3O4 H O O
OH
nanoparticleH O O C
50 µm
COOH
COOH
OH
HOOC
HOOC
COOH
OH
COOH
HOOC
HOOC OH
COOH
COOH
Enhanced Contrast
MRI
In vivo
fluorescence
imaging
Labeled Brain Tumor
Tumor site:
Enhanced image
ORMOSIL
HPPH
2
Enhance Fluorenscence Image
Targeting
Agent
Enhanced MRI
Contrast for Cancer
Enhanced In Vivo
Imaging for Drug
And Therapeutic Action
NANOMEDICINE:
Nanotechnology in
Biomedical Systems
Photodynamic Therapy
Porphyrin
hn
Porphyrin
+
O2
O2 singlet
( Localizes and
accumulates
at tumor sites )
Destroys
Cancerous Cells
Bifunctional Chromophores for Photodynamic Therapy.
Real time monitoring of drug distribution, localization and activation.
OC6H13
H3C
CH3 H3C
CH3
NH
Photosensitizer
N
C2H5
N
HN
H3C
CH3
O
HN C
O
c
P
P
S
N+ CH=CH
CH2
P P
CH CH
O
CH2CH2CH2 S OO
c
N
CH2
O
CH2CH2CH2 S ONa
O
Fluorophore for imaging
Conditions:
•Photosensitizer absorbs at a shorter wavelength than the fluorophore
•No significal energy transfer from the photosensitizer to the fluorophore
•At the excitation of fluorophore no photodynamic therapeutic action.
Collaboration: Dr. R. Pandey, Roswell Park Cancer Institute
Studies of PDT efficacy in vitro and uptake of the conjugate in vivo
Dr. R. Pandey, Roswell Park Cancer Institute
Treatm ent at 665 nm (0.6 uM)
Cell viability study
Treatm ent at 810 nm (0.6 uM)
100
Light controls
75
75
% Survival
% Survival
10 0
HPPH
50
#523
25
Excitation of photosensitizer
chromophore (665 nm),
PDT effect
Tumor
Kidney
Spleen
Brain
Heart
pancreas
Skin Adrenal
A
#523
0
0
1
2
3
0
4
Light Dose, J/cm2
0
1
2
3
Light Dose, J/cm 2
Blood
Muscle
ICG
25
Blood
Liver
Light controls
50
Muscle
Liver
Tumor
Kidney
Spleen
Brain
Heart
High
Excitation of cyanine
fluorophore (810 nm),
No PDT effect
Skin Adrenal
B
Low
Distribution of 5 in various
organ parts at (A) 48 and (B)
72 h post injection from RIF
tumor bearing mice; imaging
by fluorescence from cyanine
fluorophore
4
Two- Photon Photodynamic Therapy
Dye
2 hn
Dye+
( Two photon absorption
of light from a pulsed
laser at 800 nm)
Advantages of Up-Conversion Therapy
1. Deeper tissue penetration
2. Less collateral tissue damage
3. More Precision
+ Porphyrin
Porphyrin +
Energy transfer
O2
O2 singlet
Destroys Cancerous
Cells
Two-photon dendrimer-photosensitizer for photodynamic
therapy
1. IR Excitation
S
S
N
N
(TPA)
2. Energy Transfer
to Porphyrin
N
N
O
S
N
N
O
O
O
N
O
O
O
O
S
N
O
N NH
NH N
N
S
O
O
O
O
O
O
N
N
N
S
O
N
N
1O
3O
S
N
N
3. Singlet O2
Generation
S
In collaboration with Frechet Research Group
University of California, Berkeley
NANOPARTICLE PLATFORM FOR
PHOTODYNAMIC THERAPY
 Co-localization to Control Excitation Dynamics
 Up-conversion Photodynamics Therapy
 Multimodal Imaging Capability
 Added Targeting
 Enhanced Biodistribution
Organically Modified Silica (ORMOSIL) Nanoclinic
Encapsulating Hydrophobic Drugs for Diagnostic
Imaging and PDT
ORMOSIL Shell
HPPH
SiO2
ORMOSIL
HPPH
Targeting
Agent
20-50 nm
Schematic of HPPH doped ORMOSIL Nanoclinic
TEM image of HPPH doped ORMOSIL
nanoparticles
HPPH-ORMOSIL Nanoparticles
KB Cells
Transmission and Fluorescence Images HPPHORMOSIL nanoparticles cells and tissue
0.0309
100
Singlet Oxygen Generation
80
0.0301
60
% Cell-survival
Luminiscence Intensity (a.u.)
0.0305
0.0297
40
0.0293
20
0.0289
0
Human Tumor tissue
0.0285
1240
1250
1260
1270
1280
Wavelength (nm)
1290
1300
1310
In Vitro Cytotoxic Effect
Collocalization of a photosensitizer with heavy atom
External heavy atom effect
I
I
I
I2
I
I
I
Enhanced Intersystem Crossing
I2
I2
I
I
Enhanced Singlet Oxygen Generation
I2
I
I
I
I
I
ORMOSIL nanoparticle with
coencapsulated photosensitiser HPPH and I2
Nanoparticle shell can be modified by insertion of I.
Changes in HPPH fluorescence with the increase in I 2
concentration inside particles
Luminescence intensity, a.u.
14
12
HPPH only
1L of I2 was added to HPPH solution during preparation of particles
5L added
10L added
15L added
20L added
25L added
30L added
Excitation with 532 nm
CH3Cl solution
10
8
6
4
2
0
1200 1220 1240 1260 1280 1300 1320
Wavelength, nm
1O
580
600
620
640
660
680
700
720
740
760
generation by HPPH
manifested by 1O2 phosphorescence
780
Wavelength, nm
15
O2 Emission intensity at 1270 nm [a.u.]
180
170
160
150
140
130
120
110
100
90
80
70
60
50
40
30
20
10
0
1
Fluorescence intensity, a.u.
I2 inside nanoparticles influences on the absorption and emission of HPPH
14
13
12
11
10
9
8
7
6
5
4
3
2
1
1
Dependence of O2 generation efficiency
on concentration of I2 in ORMOSIL nanoparticles
Excitation with 532 nm
Absorbance was normalised at 532 nm
0
0
5
10
15
20
25
30
35
40
45
Quantity of I2 added during preparation
of ORMOSIL-HPPH nps, [L]
50
55
2
Gene Therapy
•Diseases (short list)
•Diabetes,
•Cystic fibrosis,
•Cancers (pancreatic, breast, prostrate, etc),
•Parkinson’s Disease
•Problems
•Genetic material susceptible to enviromental degradation
• Lack of effective in vivo delivery system.
•Viral based systems have not adequately provided answer.
•Solutions (non-viral based delivery systems)
•Liposomes
•Organic based particles (PEG, dextran, chitose, etc)
•Inorganic nanoparticles
•Silica based nanoparticles
Optically Trackable ORMOSIL Nanoparticles for Gene Delivery
FRET experiments
Encapsulated fluorescent dye And-10
(labs = 400 nm, lem = 461 nm),
DONOR
DNA stained with YOYO-1
(labs = 491 nm, lem = 509 nm)
ACCEPTOR
hn‘’
hn‘
N
HO
O
O
N
FRET
N
hn‘’’
~ 20 nm
In Vitro Uptake and Transfection of Cells by ORMOSIL/DNA
Nanoparticles
DNA delivered
into cell
nuclei
Cellular Uptake of DNA loaded ORMOSIL
nanoparticles and subsequent translocation
of DNA into the nucleus of the cell
eGFP expression
Expression of eGFP in cells
transfected with eGFP ORMOSIL
nanoparticles vector
In Vivo Transfection of Neuron
A
B
Transfection of neurons in Substantia Nigra of mouse brain (plate A) with
ORMOSIL-pEGFP. EGFP (green) is expressed in tyrosine hydroxylase
immunopositive (red) dopamine neuron (plate B).
Opportunities in Nanophotonics








Dendritic Structures for Up-conversion Lasing, Optical
Limiting and Electro-optic
Quantum-Engineering of Heirarchiacal Nanostructures
(Quantum Dot-Quantum Well, Multiple Shells)
Nanocontrol of Dynamics of Carrier Transport and Excitation
Transport
Quantum Confined Semi-Conductor: Polymner
Nanocomposties for Solar Cells and LEDs
Novel Supramolecular Templates for Self-Assemblying of
Nanostructures
Photonic Crystals Based Microcavities and Optical Circuitry
Photonically Directed Metallic Nanostructures
Molecular Electronics with Three Terminal Molecules
Opportunities in Biophotonics
• In vivo Bioimaging, Spectroscopy, and Optical
Biopsy
• Nano-Biophotonic Probes (Nanofluorophores)
• Single Molecule Biofunctions
• Multiphoton Processes for Biotechnology
• Real-Time Monitoring of Drug Interactions
• Nanomedicine
Acknowledgements
Researchers at the Institute:
Prof. E. Bergey
Prof. A. Cartwright
Prof. M. Swihart
Prof. E. Furlani
Dr. A. Kachynski
Dr. A. Kuzmin
Dr. Y. Sahoo
Dr. H. Pudavar
Dr. T. Ohulchanskyy
Dr. D. Bharali
Dr. D. Lucey
Dr. K. Baba
Dr. J. Liu
Outside Collaborators
Prof. R.Boyd
Prof. J.Haus
Prof. J M J Frechet
Prof. M. Stachowiak
Dr. A. Oseroff
Dr. R. Pandey
Dr. J. Morgan
Dr. P Dandona
DURINT/AFSOR
 Dr. Charles Lee
“Lighting the Way to
Technology through Innovation”
The Institute for Lasers, Photonics and
Biophotonics
University at Buffalo
Emerging Opportunities in
Nanophotonics and Biophotonics
www.biophotonics.buffalo.edu