cytotoxic and immunologic effects of silver nanoparticles
DESCRIPTION
Cytotoxic and Immunologic Effects of Silver Nanoparticles. Shaun Cote. What are Nanoparticles?. Any substance that has one dimension of 1-100 nanometers Commonly used nanoparticles include colloidal gold and titanium dioxide (TiO 2 ). Nano Scale. Why is this important?. - PowerPoint PPT PresentationTRANSCRIPT
![Page 1: Cytotoxic and Immunologic Effects of Silver Nanoparticles](https://reader030.vdocuments.us/reader030/viewer/2022033020/568134ad550346895d9bc4d6/html5/thumbnails/1.jpg)
Cytotoxic and Immunologic Cytotoxic and Immunologic Effects of Silver NanoparticlesEffects of Silver Nanoparticles
Shaun CoteShaun Cote
![Page 2: Cytotoxic and Immunologic Effects of Silver Nanoparticles](https://reader030.vdocuments.us/reader030/viewer/2022033020/568134ad550346895d9bc4d6/html5/thumbnails/2.jpg)
What are Nanoparticles?What are Nanoparticles?
Any substance that has one dimension of Any substance that has one dimension of 1-100 nanometers1-100 nanometers
Commonly used nanoparticles include Commonly used nanoparticles include colloidal gold and titanium dioxide (TiOcolloidal gold and titanium dioxide (TiO22))
![Page 3: Cytotoxic and Immunologic Effects of Silver Nanoparticles](https://reader030.vdocuments.us/reader030/viewer/2022033020/568134ad550346895d9bc4d6/html5/thumbnails/3.jpg)
Nano ScaleNano Scale
![Page 4: Cytotoxic and Immunologic Effects of Silver Nanoparticles](https://reader030.vdocuments.us/reader030/viewer/2022033020/568134ad550346895d9bc4d6/html5/thumbnails/4.jpg)
Why is this important?Why is this important?
““The biokinetics of NSPs are different from The biokinetics of NSPs are different from larger particles. When inhaled, they are larger particles. When inhaled, they are efficiently efficiently deposited in all regions of the deposited in all regions of the respiratory tractrespiratory tract; they ; they evade specific defense evade specific defense mechanismsmechanisms; and they can ; and they can translocate out of translocate out of the respiratory tractthe respiratory tract via different pathways and via different pathways and mechanisms (endocytosis and transcytosis).”mechanisms (endocytosis and transcytosis).”
Nanotoxicology: An Emerging Discipline Evolving from Studies of Ultrafine Particles.Environmental Health Perspectives, Oberdörster et al. 113(7), July 2005.
![Page 5: Cytotoxic and Immunologic Effects of Silver Nanoparticles](https://reader030.vdocuments.us/reader030/viewer/2022033020/568134ad550346895d9bc4d6/html5/thumbnails/5.jpg)
Downsides to Silver NanoparticlesDownsides to Silver Nanoparticles
Possible negative effects due to the Possible negative effects due to the chemistry of small moleculeschemistry of small molecules
![Page 6: Cytotoxic and Immunologic Effects of Silver Nanoparticles](https://reader030.vdocuments.us/reader030/viewer/2022033020/568134ad550346895d9bc4d6/html5/thumbnails/6.jpg)
How they are MadeHow they are Made
Metal salt Metal salt solutions are solutions are boiled and then boiled and then reduced via a reduced via a reducing agentreducing agent
![Page 7: Cytotoxic and Immunologic Effects of Silver Nanoparticles](https://reader030.vdocuments.us/reader030/viewer/2022033020/568134ad550346895d9bc4d6/html5/thumbnails/7.jpg)
Silver Nanoparticles in the MarketSilver Nanoparticles in the Market
SamsungSamsung Washing machine, refrigerator and air Washing machine, refrigerator and air
conditionersconditioners
NanohorizonsNanohorizons Smart Silver – silver nanoparticles intertwined Smart Silver – silver nanoparticles intertwined
with fabric to repel odor and microbeswith fabric to repel odor and microbes
Aigon Aigon SilverClene anti-microbial cleaning solutionSilverClene anti-microbial cleaning solution
![Page 8: Cytotoxic and Immunologic Effects of Silver Nanoparticles](https://reader030.vdocuments.us/reader030/viewer/2022033020/568134ad550346895d9bc4d6/html5/thumbnails/8.jpg)
Project GoalsProject Goals
To explore the effects of silver To explore the effects of silver nanoparticles by testing the:nanoparticles by testing the: Cytotoxicity on established cell lines:Cytotoxicity on established cell lines: Production of specific immune moleculesProduction of specific immune molecules
![Page 9: Cytotoxic and Immunologic Effects of Silver Nanoparticles](https://reader030.vdocuments.us/reader030/viewer/2022033020/568134ad550346895d9bc4d6/html5/thumbnails/9.jpg)
TestingTesting
1)1) MTT Assay – tests for metabolic activityMTT Assay – tests for metabolic activity
2)2) TNF-TNF-αα ELISA – Antibody test for TNF- ELISA – Antibody test for TNF-αα secretion from cellssecretion from cells
3)3) RT-PCR – Measure of RNA activity RT-PCR – Measure of RNA activity
![Page 10: Cytotoxic and Immunologic Effects of Silver Nanoparticles](https://reader030.vdocuments.us/reader030/viewer/2022033020/568134ad550346895d9bc4d6/html5/thumbnails/10.jpg)
Cell Lines TestedCell Lines Tested
RAW 264.7 – Mouse macrophageRAW 264.7 – Mouse macrophage Jurkat – Human T cellsJurkat – Human T cells WHTBF-6 – Human lung fibroblastWHTBF-6 – Human lung fibroblast
![Page 11: Cytotoxic and Immunologic Effects of Silver Nanoparticles](https://reader030.vdocuments.us/reader030/viewer/2022033020/568134ad550346895d9bc4d6/html5/thumbnails/11.jpg)
MTT AssayMTT Assay Cells are cultured, then treated with silver Cells are cultured, then treated with silver
nanoparticlesnanoparticles MTT is added, and blue crystals are MTT is added, and blue crystals are
formed if the cells are viableformed if the cells are viable Correlation between concentration of Correlation between concentration of
nanoparticles andnanoparticles andviability of the cellsviability of the cells
![Page 12: Cytotoxic and Immunologic Effects of Silver Nanoparticles](https://reader030.vdocuments.us/reader030/viewer/2022033020/568134ad550346895d9bc4d6/html5/thumbnails/12.jpg)
MTT Assay - ResultsMTT Assay - ResultsRAW 264.7 Cell Line
0.00%
20.00%
40.00%
60.00%
80.00%
100.00%
120.00%
0 0.5 1 1.5 2 2.5 3 3.5 4 4.5
Concentration
Per
cen
t V
iab
ility
Jurkat Cell Line
-20.00%
0.00%
20.00%
40.00%
60.00%
80.00%
100.00%
120.00%
0 0.5 1 1.5 2 2.5 3 3.5 4 4.5
Concentration
Per
cen
t V
iab
ility
WHTBF-6 Cell Line
-20.00%
0.00%
20.00%
40.00%
60.00%
80.00%
100.00%
120.00%
140.00%
0 0.5 1 1.5 2 2.5 3 3.5 4 4.5
Concentration
Per
cen
t V
iab
ility
![Page 13: Cytotoxic and Immunologic Effects of Silver Nanoparticles](https://reader030.vdocuments.us/reader030/viewer/2022033020/568134ad550346895d9bc4d6/html5/thumbnails/13.jpg)
TNF-TNF-αα ELISA ELISA
Test based on antibody capture of TNF-Test based on antibody capture of TNF-αα, , a protein molecule essential in triggering a protein molecule essential in triggering an immune responsean immune response
Measured OD of sample wells compared Measured OD of sample wells compared to a standard curveto a standard curve
![Page 14: Cytotoxic and Immunologic Effects of Silver Nanoparticles](https://reader030.vdocuments.us/reader030/viewer/2022033020/568134ad550346895d9bc4d6/html5/thumbnails/14.jpg)
TNF-TNF-αα ELISA Results ELISA Results
0
50
100
150
200
250
300
350
400
C C-2.0 C-4.0 0.125 0.25 0.5 1.0 2.0 4.0 LPS
>60,000
Concentration Concentration of silver of silver nanoparticles nanoparticles versus versus relative relative concentration concentration of TNF-of TNF-αα
![Page 15: Cytotoxic and Immunologic Effects of Silver Nanoparticles](https://reader030.vdocuments.us/reader030/viewer/2022033020/568134ad550346895d9bc4d6/html5/thumbnails/15.jpg)
RT-PCRRT-PCR
A method of amplifying target RNA to A method of amplifying target RNA to determine how silver nanoparticles affect determine how silver nanoparticles affect gene expression of specific cytokinesgene expression of specific cytokines
![Page 16: Cytotoxic and Immunologic Effects of Silver Nanoparticles](https://reader030.vdocuments.us/reader030/viewer/2022033020/568134ad550346895d9bc4d6/html5/thumbnails/16.jpg)
Results of RT-PCRResults of RT-PCR
Only tested GAPDH, a housekeeping Only tested GAPDH, a housekeeping genegene
Further tests include the cytokines TNF-α, Further tests include the cytokines TNF-α, IL-6, and IL-1β IL-6, and IL-1β
![Page 17: Cytotoxic and Immunologic Effects of Silver Nanoparticles](https://reader030.vdocuments.us/reader030/viewer/2022033020/568134ad550346895d9bc4d6/html5/thumbnails/17.jpg)
Further StudiesFurther Studies
Continuing RT-PCR studies on other Continuing RT-PCR studies on other cytokinescytokines
![Page 18: Cytotoxic and Immunologic Effects of Silver Nanoparticles](https://reader030.vdocuments.us/reader030/viewer/2022033020/568134ad550346895d9bc4d6/html5/thumbnails/18.jpg)
AcknowledgementsAcknowledgements
Dr. Ah-Kau Ng and LabDr. Ah-Kau Ng and Lab Theresa KnightTheresa Knight Jennifer WalkerJennifer Walker SURF programSURF program