Effects Of Inorganic Bovine Bone Treated With Low Temperature Argon-oxygen Plasma On MC3T3-E1 Cells

Objective:As a kind of artificial bone substitute,inorganic bovine bone has been applied in the field of oral cavity due to its good biocompatibility and bone conduction function,but it has defects such as poor osteogenic effect and incomplete bone remodeling,etc.This experiment attempts to find a simple,safe and efficient surface treatment method to enhance the osteogenic effect of inorganic bovine bone.The main purpose of this study was to study the adhesion,proliferation and differentiation of mouse embryonic osteoblast MC3T3-E1 on inorganic bovine bone treated with low temperature argon-oxygen plasma.Methods : The inorganic bovine bones were irradiated with ultraviolet lamp for disinfection for 2 hours.After the disinfection,the bones were prepared into cylindrical bones with a diameter of 5mm and a height of 2mm using sterile annulus cutter and sterile medical blade.A low temperature argon-oxygen plasma was generated with argon +5% oxygen as the output gas for surface activation of inorganic bovine bones.Inorganic bovine bones treated with low temperature argon-oxygen plasma were used as the experimental group,while inorganic bovine bones not treated were used as the control group.The surface morphologies of the experimental group and the control group were observed by scanning electron microscopy(SEM),and the surface element composition of the experimental group and the control group was detected by X-ray photoelectron spectroscopy(XPS).MC3T3-E1 cells were inoculate on the material surface of the experimental group and the control group,respectively,and the adhesion morphology of MC3T3-E1 cells on the material surface of the two groups was observed by scanning electron microscopy.Proliferation changes of the cells in the two groups were detected by CCK-8 method on day 1,3 and 5,and the differentiation state of the cells in the two groups was detected by alkaline phosphatase(ALP)method on day 7 and day 14.Results:Under scanning electron microscope,the surface morphology of inorganic bovine bone in the experimental group and the control group showed no significant change,showing rough surface.X-ray photoelectron spectroscopy analysis of the material element composition of the experimental group and the control group showed that compared with the control group,the surface carbon(C)element in the experimental group decreased,while the oxygen(O)element,calcium(Ca)element and phosphorus(P)element increased.Under electron microscope,the cells in the experimental group adhered more fully to the surface of the material,and the cells extended pseudopodia,while the cells in the control group were spherical.The cell proliferation test results showed that the cell proliferation in the 1,3,5 day argon oxygen group was significantly higher than that in the control group,and the difference was statistically significant(P < 0.05).The cell differentiation results showed that there was no significant difference in ALP activity between the experimental group and the control group at 7 days,but the ALP activity of the experimental group was higher than that of the control group at 14 days,and the difference was statistically significant(P < 0.05).Conclusion: Inorganic bovine bone treated with low temperature argon-oxygen plasma can promote the adhesion,proliferation and differentiation of MC3T3-E1,and the low temperature argon-oxygen plasma will not destroy the surface morphology and element composition of inorganic bovine bone.This experiment provides a new idea for the surface modification of artificial bone substitute.