Discipline: Biological Sciences
Subcategory: Cancer Research
DeQuaysha Greene - Jarvis Christian College
Co-Author(s): Shakhawat Bhuiyan, Jarvis Christian College, Hawkins, Texas
Nanoscience and nanotechnology have evolved in a way that we now can diagnose, treat and prevent numerous diseases in all aspects of human life. Silver nanoparticles (AgNP) have unique optical, electrical, and thermal properties and are being incorporated into products that range from photovoltaics and biological and chemical sensors. Silver nanoparticles are one of the most important nanomaterials that are involved in the advance biomedical applications. In the present research, silver nanoparticles were synthesized from Spinacia oleracea (Spinach) and tested on human lymphoblastoid TK6 cells. We hypothesized whether synthesized silver nanoparticles would increase the apoptosis of lymphoblastoid TK6 cells. To make silver nanoparticles, 20g of spinach was boiled in 200 ml of distilled water to making an extract. After filtration, 1mM of silver nitrate solution was added to spinach extract and left at room temperature for 24h. The size of the nanoparticle was measured at 50 d/nm using a UV spectrophotometer. The human lymphoblastoid Tk6 cells (American Type Culture Collection) were grown in RPMI-1640 medium supplemented with 10% fetal bovine serum and 1.0% penicillin/streptomycin (Invitrogen) at 370°C in 5% CO2/95% air for 24h, 48h, and 72h. To determine the effects of silver nanoparticles, the TK6 cells were treated with different concentrations of AgNP for 3 days. The viability of the treated and untreated cells was measured by Trypan blue using a Hemocytometer. The results showed that the cells treated at various concentrations of AgNP (1, 3, 10 and 20 µM) showed the highest killing (100%) at 20µM of AgNP after 24h of treatment. After 3 days of treatment with silver nanoparticles, cells were harvested at -20℃ freezer and extracted proteins from treated cells. The protein concentrations were measured at 562 nm by BCA method using Multiscan FC. The results showed that total protein concentrations were reduced as concentrations of nanoparticles increased. Furthermore, the western blot analysis of P42/44 MAP kinase protein showed the decreased in phosphorylation as compared to control. Our results showed that the synthesized silver nanoparticles from spinach extract significantly inhibited the growth of human lymphoblastoid TK6 cells.
Funder Acknowledgement(s): National Science Foundation Grant (J-Cure Program)
Faculty Advisor: Shakhawat Bhuiyan, firstname.lastname@example.org
Role: I have completed all parts of the research including cell culture, maintenance of cells, extraction of silver nanoparticles from spinach, treatment of cells with various concentrations of silver nanoparticles, extraction of protein from cells, and perform western blotting analysis.