Transcript Pui Kam Tse - Harlem Children Society

Detecting and Curing Skin Cancer by using
Gold Nanoparticles with Photosensitizer
Pui Kam Tse
Mentor :Dr. Henry Du
Stevens Institute of Technology
Key Terms
 Cancer: cancer develops when cells in the body begin to
divide and reproduce abnormally and have the potential
to spread through the body.
 Nanoparticle: a nanoparticle is a microscopic particle
whose size is measured in nanometer (nm).
 Photodynamic Therapy (PDT): the use of light to
induce reaction in the body which helps treating
diseases in patients.
 Photosensitizer / Photosensitizing Agent: a drug uses
in PDT.
 Surface-Enhanced Raman Spectroscopy (SERS): it
is a sensitive vibrational spectroscopy that gives
structural information on the molecule and its local
interactions
Introduction
 There are three types
of skin cancer
 Basal cell carcinoma
(75%)
 Squamous cell
carcinoma (20%)
 Melanoma cell
carcinoma (5%)
…Continued
How does PDT work?
…Continued
The roles of gold nanoparticles and photosensitizer
Gold nonoparticles
-have a greater attraction to cancer cells
than to healthy cells
-raman enhance the cells because gold
nanoparticles are very good at scattering
and absorbing light
photosensitizer
-kills cancer cells by laser excitation
Objectives
Synthesize gold nanoparticles
Apply gold nanoparticles with
photosensitizer to healthy cells and skin
cancer cells then observe what will
happen to both of them
Enhance the PDT effect and drug delivery
Procedures
 synthesize gold nanoparticles
solution 1
solution 2
solution 3
0.5ml citrate
+4.5ml H2O
2.5ml citrate
+ 2.5ml H2O
5ml citrate
5ml HAuCl4
5ml HAuCl4
5ml HAuCl4
Total: 10ml
Total: 10ml
Total: 10ml
Citrate: HAuCl4
1:1
…Continued
 Heat up the solutions by UV light for at least
one hour.
Testing the Sizes of Gold Nanoparticles
Zetasizer
Sub-culturing Cells (Fibroblasts)
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suck out the media from the flask
cover all the cells in the flask with 10 ml trypsin
Incubate it for about 3-5minutes
suck out the trypsin from the flask
put in 8 ml DMEM media to the flask
repeat the steps of sucking in and out to make sure
cells do not stick on the wall of the flask
transfer the cell suspension to a sterile centrifuge tube for 3
minutes
remove the supernatant and add new media to the pellet
place the cell suspension into 6 well places
Incubate the well plates for 1 hour and 30 minutes
add gold and silver nonoparticles to the well plates (Experiment 2,
add gold nanoparticles with photosensitizer -ALA (Deltaaminolevulinic acid) to the well plates.)
Incubate the well plates for 24 hours
(ALA)
Fibroblasts with Gold and Silver Nanoparticles
Before
After
Fibroblasts + Au
fibroblasts
Fibroblasts + Ag
MTT (Dimethylthiazol) Test
 Suck out the media from well
plates
 Put in 500μl HBSS in each well
plate
 Suck out the HBSS
 Put in 500μl MTT
 Incubate the well plates for 1 hour
 Suck out the MTT
 Put in 500μl DMSO media in each
well plate
 Cover the well plates and keep
shaking for a few minutes
 Transfer the solution into a 96 well
plates
 Add 100μl DMSO media to two
blank well plates as control
Plate reader
Procedures
Surface-Enhanced Raman Spectroscopy (SERS)
Cells
SERS
Results
Sizes of Gold Nanoparticles
Solution 1
Solution 2
Solution 3
27.2nm
29.0nm
39.3nm
…Continued
MTT Test 1(fibroblasts, Au & Ag nanoparticles)
0.4
0.35
0.3
0.25
0.2
0.15
0.1
0.05
0
Ag
Au
Ctrl
…Continued
MTT Test
Ct
r
Au
+1
00
pp
m
Series1
Au
+1
00
0p
pm
Au
+1
00
00
pp
m
0.45
0.4
0.35
0.3
0.25
0.2
0.15
0.1
0.05
0
SERS Results
100000
90000
80000
10000ppm
10000ppm+Ag
70000
60000
50000
40000
30000
20000
10000
0
0
500
1000
1500
2000
2500
3000
3500
4000
Conclusion
 Different concentrations of gold nanoparticles have
different colors and sizes.
 MTT Test 1 shows the survival rate of cells after adding
silver nanoparticles compares to survival rate after
adding gold nanoparticles are very close.
 Results from MTT Test 2 show that most of the cells
survive after adding nanoparticles with photonsensitizerALA; therefore, we conclude that without light source
ALA is nontoxic to healthy cells.
 Raman signal of ALA increases dramatically after
adding silver/gold nanoparticles.
Future goals
 apply gold nanoparticles and photosensitizer to
skin cancer cells and see if gold nanoparticles
with photosensitizer can kill the cancer cells
efficaciously.
 Raman spectrum responses from healthy and
diseased cells.
 Detect skin cancer cells more easily and
precisely through the use of gold nanoparticles.
References
 Kneipp, K.2002.Surface-Enhanced Raman Spectroscopy
in Single Living Cells Using Gold Nanoparticles. Society
for Applied Spectroscopy. vol. 56, number 2:pgs 150154.
 Dougherty, T. J and Levy, J .G. Biomedical Photonics
Handbook .38-1-13-14.
 www.cancer.org
 www.photochembgsu.com/applications/therapy.html
 www.answers.com
 http://www.cancercouncil.com.au/editorial.asp?pageid=5
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Acknowledgements
 Dr. Sat Bhattacharya
 MSKCC
 Harlem Children Society
 Dr. Henry Du
 Dr.Wong
 Malcolm
 Yun Han
 Stevens Institute of Technology
 All the people I forgot to mention