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Synthesis and Evaluation of the ability to inhibit gastric cancer cells of AgNPs-Croton tonkinensis Gagnep.

Description: Novelty of the study The combination of a herbal medicinal plant - Croton tonkinensis Gagnep. and silver nanoparticles Practicality of the study AgNPs-Croton tonkinensis Gagnep. complexes are silver nanosystems synthesized by biological method, so it is less toxic than many other anti-cancer drugs. This complication, if successfully tested, could be directed to a treatment for gastric cancer. Pharmacology Ability of Croton tonkinensis Gagnep. Croton used its’leaves for healing purpose, and has a bitter taste to help with heat. Croton contains substances with anti-inflammatory and antibacterial properties In the leaves of the croton contains 4 substances: Flavonoids, alcaloid, tannin, polyphenols that prevent and destroy Helicobacter pylori bacteria. One of the main ingredients in the leaves of the Diterpenoid plant is the diterpenoid that has been shown to be toxic to cancer cells. CONCLUSIONS Successfully fabricated AgNPs-Croton tonkinensis Gagnep complex from AgNO3 salt and extracts

Synthesis method of AgNPs-Croton tonkinensis Gagnep. Step 1: Prepare 5mM AgNO3 salt solution, stir well with magnetic stirring. Step 2: Transfer 42ml 5mM AgNO3 solution to a 50ml measuring cup and place on the magnetic stirrer at 1000C, 600 rounds for 10 minutes. Step 3: Add 2ml, 5ml or 8ml of croton’s extract, add water to the total volume of 50ml, stir for 10 minutes. Step 4: Lower the temperature to 60 ° C, stir for 30 minutes, observe the color change. Step 5: Cool the solution and measure UV-VIS spectrum (250 - 800 nm). Step 6: Store the solution at 4 ° C Method of culturing and processing cells with AgNPs-Croton tonkinensis Gagnep complex. Prepare the culture medium. Cell culture: Put cells into 96-well plate, add 100µl of culture medium, put into CO2 culture cabinet at 37 ° C, 5% CO2. After 48 h of culture, cells were treated with AgNPs-Croton tonkinensis Gagnep complex. at concentrations of 0.05; 0.1; 0.2; 0.5 and 1 μg / ml, the control samples were extracts with treatment time of 24 hours and 48 hours. Method of cell survival assessment (MTT method) The yellow MTT salt is produced by active metabolic cells by the dehydrogenase enzyme, giving a balanced amount of NADH and NADPH reduction products. The result is a soluble purple endothelial formazan molecule that is quantified by a spectrophotometer Flow cytometry analysis method of apoptosis Prepare a dye solution of Propidium Iodide (PI). Cell processing process: Cell collection, centrifuge to remove the solution, add 200µl of 75% ethanol, keep at -200C. Cell staining process: Step 1: Centrifuge the tube containing the fixed cells at 1300 rounds for 5 minutes. Step 2: Remove translation. Step 3: Add 100 µl PBS. Step 4: Centrifuge remove fluid at 1300 rounds for 5 minutes. Step 5: Add 100 µl of PI dyeing solution. Step 6: Incubate for 30 minutes at 4 ° C in the dark, analyzed by Flow cytometry. DAPI dye method to evaluate the morphology of the cell nucleus Process of staining cells with DAPI: Step 1: Remove the culture medium. Step 2: Wash the surface of the culture dish with 50µL of PBS 1X. Step 3: Remove PBS, add 50µL of paraformaldehyde 4% solution, leave at room temperature for 10 minutes. Step 4: Remove the paraformaldehyde and wash the surface of the culture plate 2 times with PBS 1X. Step 5: Add 70µL of DAPI nucleus dye solution (0.1 μg / ml). Step 6: Observe under reverse fluorescence microscope and record images Analytical method of ROS production in cells Step 1: After 24 h culture, the cells were treated with AgNPs-Croton tonkinensis Gagnep. complex for 48 hours at different concentrations. Step 2: Remove the cell culture medium. Step 3: Wash the surface of the cultured well with PBS 1X. Step 4: Fix with 4% paraformaldehyde at room temperature for 5 minutes. Step 5: Wash the surface of cultured wells 2 times with PBS 1X. Step 6: Add DCF-DA solution and incubate for 30 minutes in a cell culture cabinet of 5% CO2. Step 7: Wash the surface of cultured wells 2 times with PBS 1X. Step 8: Record cell images using a fluorescence microscope. Result Structure analysis results of nano silver (AgNPs) : Nanoparticles are synthesized using X-ray diffraction spectroscopy (XRD) to confirm the particles are silver and to obtain structural information Analysis results of transmission electron microscopy of AgNPs-Croton tonkinensis Gagnep complex : Analysis results of transmission electron microscope image (TEM) of the AgNPs-Croton tonkinensis Gagnep complex. shows that the particles are spherical, evenly distributed, with the most common average size of 14nm. Effects of complex AgNPs-Croton tonkinensis Gagnep. to speed up the growth of gastric cancer cells : Gastric cancer cells were cultured on culture plates for 24 hours, then treated with the AgNPs-Croton tonkinensis Gagnep. complex at all concentrations and observations, assess the effect after 24 hours and 48 hours of treatment. The proliferation decreased markedly with increasing concentration of the above co

Organisation: RESEARCH INSTITUTE OF CREATIVE EDUCATION

Innovator(s): Nguyen Trung Chien, Phan Ba Thai Duong, Pham Thu Huong, Cao Dieu Ngan, Ngo Thi Thu Thuy

Category: Biotechnology

Country: Vietnam

Gold Award