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Cytotoxic and Immunologic Effects of Silver Nanoparticles

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?.

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Cytotoxic and Immunologic Effects of Silver Nanoparticles

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  1. Cytotoxic and Immunologic Effects of Silver Nanoparticles Shaun Cote

  2. What are Nanoparticles? • Any substance that has one dimension of 1-100 nanometers • Commonly used nanoparticles include colloidal gold and titanium dioxide (TiO2)

  3. Nano Scale

  4. Why is this important? • “The biokinetics of NSPs are different from larger particles. When inhaled, they are efficiently deposited in all regions of the respiratory tract; they evade specific defense mechanisms; and they can translocate out of the respiratory tract via different pathways and mechanisms (endocytosis and transcytosis).” Nanotoxicology: An Emerging Discipline Evolving from Studies of Ultrafine Particles. Environmental Health Perspectives, Oberdörster et al. 113(7), July 2005.

  5. Downsides to Silver Nanoparticles • Possible negative effects due to the chemistry of small molecules

  6. How they are Made • Metal salt solutions are boiled and then reduced via a reducing agent

  7. Silver Nanoparticles in the Market • Samsung • Washing machine, refrigerator and air conditioners • Nanohorizons • Smart Silver – silver nanoparticles intertwined with fabric to repel odor and microbes • Aigon • SilverClene anti-microbial cleaning solution

  8. Project Goals • To explore the effects of silver nanoparticles by testing the: • Cytotoxicity on established cell lines: • Production of specific immune molecules

  9. Testing • MTT Assay – tests for metabolic activity • TNF-α ELISA – Antibody test for TNF-α secretion from cells • RT-PCR – Measure of RNA activity

  10. Cell Lines Tested • RAW 264.7 – Mouse macrophage • Jurkat – Human T cells • WHTBF-6 – Human lung fibroblast

  11. MTT Assay • Cells are cultured, then treated with silver nanoparticles • MTT is added, and blue crystals are formed if the cells are viable • Correlation between concentration of nanoparticles and viability of the cells

  12. MTT Assay - Results

  13. TNF-α ELISA • Test based on antibody capture of TNF-α, a protein molecule essential in triggering an immune response • Measured OD of sample wells compared to a standard curve

  14. TNF-α ELISA Results • Concentration of silver nanoparticles versus relative concentration of TNF-α

  15. RT-PCR • A method of amplifying target RNA to determine how silver nanoparticles affect gene expression of specific cytokines

  16. Results of RT-PCR • Only tested GAPDH, a housekeeping gene • Further tests include the cytokines TNF-α, IL-6, and IL-1β

  17. Further Studies • Continuing RT-PCR studies on other cytokines

  18. Acknowledgements • Dr. Ah-Kau Ng and Lab • Theresa Knight • Jennifer Walker • SURF program

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