Study On Biological Properties Of Antibacterial Bone Repair Material HAPw/nmZnO-nmCaO

[Objectives]In this experiment,based on the previous study of the research group,HAPw was modified by sol-gel method to prepare HAPw/nmZO-nmCaO composite material.The cytotoxicity and cell proliferation of the composite materials were further investigated on human SV40 transfected osteoblasts and mouse embryonic fibroblasts,to understand the biological properties of the composite materials.[Materials and Methods]?HAPw was modified via sol-gel method using calcium nitrate(raw material),zinc nitrate(raw material)and PEG6000(dispersant),to obtain HAPw/nmZnO-nmCaO composite materials which were characterized by XRD and SEM;?Human SV40 transfected osteoblasts and mouse embryonic fibroblasts were used as experimental models in vitro.Human SV40 cells at good logarithmic phase were transfected into osteoblasts and mouse embryonic fibroblasts which were treated using different concentrations of HAPw/nmZnO-nmCaO(0,0.1,1,10mg/ml)post-incubation 24h,48h and 72h,subsequently,CCK-8 assay was used to observe the proliferation ability of human SV40 transfected osteoblasts and mouse embryonic fibroblasts and the results were measured by OD value;?Additionally,the effect of HAPw/nmZnO-nmCaO on the expression of ALP activity in osteoblasts transfected with human SV40 was detected by real-time fluorescence quantitative PCR.Additionally,the of Runx2 and TGF-P mRNA in human SV40-transfected osteoblasts were monitored by real-time fluorescent quantitative PCR Variety.[Res?lts]?Under the optimal modification process,including the main phase hydroxyapatite,the characteristic peaks of CaO and ZnO appeared on the diffraction pattern of the modified material,which did not affect the peak position of the main phase HAP;?When concentrations of HAPw/nmZnO-nmCaO(0,0.1,1,10 mg/ml)acted on human SV40 transfected osteoblasts and mouse embryonic fibroblasts for 24h,48h,and 72h,the cells,respectively.The number of osteoblasts Increased significantly.And the OD values of all groups increased,especially at the concentration of 10 mg/ml.Compared with the blank control group,the experimental group is statistically significant(P<0.05).There is no significant difference between the experimental group and the experimental control group(P>0.05);?On the other hand,when HAPw/nmZnO-nmCaO(0,0.1,1,10 mg/ml)acted on human SV40-transfected osteoblasts for 72 h.Compared with the control group,HAPw/nmZnO-nmCaO could promote ALP viability in all groups.There is statistically significant(P<0.05).In addition,when different concentrations of HAPw/nmZnO-nmCaO(0,0.1,1,10 mg/ml)acted on human SV40-transfected osteoblasts for 72 h.The resuLts of PCR assay showed that the expression of Runx2 and TGF-? mRNA in human SV40 transfected osteoblasts were relatively increased,and the gene expression at 10 mg/ml increased significantly.There is a statistical difference between the control group and the experimental group(P<0.05).[Conclusions]?Using calcium nitrate and zinc nitrate as raw materials,PEG6000 as a dispersant,HAPw was modified by sol-gel method,and successfully prepared antibacterialbone repair material(HAPw/nmZnO-nmCaO);?HAPw/nmZnO-nmCaO has no cytotoxicity to human SV40 transfected osteoblasts and mouse embryonic fibroblasts and can promote the cells proliferation;?HAPw/nmZnO-nmCaO can promote.ALP activity in human SV40 transfected osteoblasts and have osteogenic function.This material may promote bone formation by increasing the expression of Runx2 and TGF-p mRNA in osteoblasts by human SV40.