In-vitro study of cytotoxicity and genotoxicity of nanoamorphus calcium phosphate(NACP) in macrophage RAW264 cell line

ObjectiveThe biological assessment of cultured RAW264 macrophage to nano amorphous calcium phosphate particles by analyzing of cytotoxicity and genotoxicity tests.MethodsIn this study Nanoamorphus Calcium Phosphate particles produced by sol-gel method in Iran polimer center, then particle size and hemogenicity was analysed by XRD (X ray Diffraction). Cytotoxicity of nanoparticles was determinatedon mouse RAW264 macrophage. The cells cultured in 37 c in DMEM medium with 10% fetal calf serum and 1% antibiotic and antifangal under 5% CO2 atmosphere. the grown cellsin 96 part plates with concentration of 10000 cells in each part with 200 micro liter medium cultured in 72 hours, after these time medium removed and the second medium added to cells which contained two different concentrations of nano particle(200&400μg/ml) and the cells which didn t have any nano particle consider as control group. Then the cells was ancubated for 24 hours and the cell vitality assessed by MTT assay (calorimetric method use for counting vital cells), cell morphology,cell apoptosis analysis by Annexin V(method for counting apoptotic cells with flowcytometery) ;were done in BU-Ali immunohistochemical research center.ResultsResults reached from Annexin V assay show that apoptosis increase in macrophages with 200,400μg NACP but it is a few to count; and after 24 hours of ancubation, most viable cell were in 200&400μg group. Cytotoxicity measured by MTT assay showed that cell viability in three group contains of control,200 μg and 400 μg were 100,107,103 percents and so most living cell were in 200 μg and then 400 μg which were more than control group; exactly similar to result of Annexin V assay that shows that Nanoamorphus Calcium Phosphate has no toxicity.ConclusionNACP has no cytotoxicity and genotoxicity, so it can be used as non-toxic and benefical material for clinical use.

کلیدواژه ها:

Cytotoxicity ، Genotoxicity ، Nanoamorphus calcium phosphate