The Experiment Of Metal Ion Stimulate Human Mononuclear Cell Apoptosis And Produce TNF-α

Objective:To observe the mononuclear cell release TNF-αand apoptosis after stimulated by cobalt ion(Co2+) and chromium ion (Cr3+). Then discuss the mechanism of artificial joint wear production metal ion cobalt(Co2+) and chromium(Cr3+) on aseptic loosening.Method:CoCl2 powder and CrCl3 powder dissolve asepsis inject water, confirming asepsis before use. Using lymphofolt and Percoll gradient centrifugation acquire mononuclear cell from peripheral blood , mononuclear has been cultured in the 24 holes culture plate, 4×106/hole. This experiment is divided into 3 groups: group A is mononuclear cell culture alone as negative; group B is mononuclear cell culture with 10ppm Co2+; group C is mononuclear cell culture with 500ppm Cr3+. Assessing the production of TNF-αby enzyme-linked immunoabsorbent assay at and detecting the apoptosis by flow cytometer at 12,24,48-hour after co-cultured respective.Result:①12h experimental group supernatant liquid TNF-αvalue is obviously higher than control group, the TNF-αvalue of 24h experimental group Co2+ and Cr3+ is obviously higher than other time experimental group.②the cell apoptosis rate of experimental group at every time point is higher than control group, the cell apoptosis rate of 48h experimental group is obviously higher than other time experimental group (P<0.05).Conclusion:Co2+ and Cr3+ ions can stimulate human mononuclear cell release TNF-α, wihich is time-dependent, arrive to peak at 24h. Co2+ and Cr3+ can induce human mononuclear cell apoptosis, and apoptosis rate is heighten with the time. Co2+ and Cr3+ can stimulate human mononuclear cell release TNF-αand induce human mononuclear cell apoptosis which result in periprosethetic osteolysis. Then cuase prosethetic aseptic loosening.