Detecting and Curing Skin Cancer by using Gold Nanoparticles

with Photosensitizer

Pui Kam TseMentor :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?

…ContinuedThe 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 nanoparticlesApply gold nanoparticles with photosensiti

zer 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) 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 (Delta-aminolevulinic acid) to the well plates.)

Incubate the well plates for 24 hours (ALA)

Fibroblasts with Gold and Silver Nanoparticles Before After

Fibroblasts + Au

Fibroblasts + Ag fibroblasts

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 controlPlate reader

ProceduresSurface-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

0.05

0.1

0.15

0.2

0.25

0.3

0.35

0.4

Ag Au Ctrl

…Continued

MTT Test

00.05

0.10.15

0.20.25

0.30.35

0.40.45

Series1

SERS Results

0

10000

20000

30000

40000

50000

60000

70000

80000

90000

100000

0 500 1000 1500 2000 2500 3000 3500 4000

10000ppm

10000ppm+Ag

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 photonsensitizer-ALA; 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 150-154.

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