Research Biocompatibility Of Osteoblasts With Nano Hydroxyapatite/Artificial Bone Collagen (NHAC) Adding Vascular Endothelial Growth Factor (VEGF)

Objective:Choose the osteoblast cultured in vitro and Nano Hydroxyapatite/Artificial bone Collagen (NHAC) to be cultured together with Vascular Endothelial Growth Factor (VEGF),evaluating the osteocytes compatibility of composite materials from the area of cytology, while comparing with the allogeneic bone and bone cement.To provid the basis for clinical application.Method:Culture the osteoblast in vitro using the 106 generation of MG63 cells, also take the NHAC/VEGF,allogeneic bone and bone cemen these three materials as the basement. Test the cell proliferation activity using four methyl azo thiazole salt trace enzyme reaction colorimetric method (MTT). And assay the total protein content using coomassie brilliant blue method. Compared with NHAC/VEGF, allogeneic bone and bone cement three materials for the influence on the cell activity.Assay the mRNA transcription of ALP and osteocalcin (OC)during the culture of osteoblast through Revorse Transcriptase-Polymerase Chain Reaction(RT-PCR).Estimate inorganic calcium quantitatively secreted by osteoblasts of those three materials,taking the calcium standard colorimetric method.To analysis of osteocytes compatibility and any significant difference of that between groups. Results:1,The results of dynamic measurement in MTT method show that the number of cell with each material increases with the extension of culture time, the absorbance values of cells between groups at all time points are significantly different.2,About the protein content of osteoblasts in different material on the 4th day,which in the allogeneic bone group is more than other groups, in the NHAC/VEGF group is more than control and bone cement group,and threre is little difference between control and bone cement group. As the Cultivating prolonged, protein content are increasing. Protein content in each groups increases obviously on the 7th day, comparing to the 10th day, no obvious difference exists.3,Determination of osteoblast differentiation capacity:ALP has expressed in each groups after osteoblasts in three kinds of materials cultured 7 days.On the relative level of ALP mRNA expression, allogeneic bone group>NHAC/VEGF group>bone cement group,with significant differences between the groups(P<0.05),OCN in each doesn't express.After 14 days of cultuer, on the relative level of ALP mRNA expression, there is no significant differences between allogeneic bone group and NHAC/VEGF ones,but both them are more than bone cement group. OCN mRNA doesn't espress in bone cement group,but a little expression in allogeneic bone group and NHAC/VEGFALP has expressed in each groups after 21 days.On the relative level of ALP mRNA expression, bone cement group>NHAC/VEGF group>allogeneic bone group,with significant differences between the groups(P<0.05).OCN mRNA in three groups have expressed, which in allogeneic bone group and NHAC/VEGF group is more than bone cement group, with no significant differences of its relative expression level between the two groups (P>0.05), but with obvious difference comparing with bone cement group(P<0.05).4,determination of inorganic calcium of Osteoblasts in different materials:With the increase of training time, the calcium content of the cell in each group significantly increased, the level of inorganic calcium secreted by cell in allogeneic bone group is more than other groups throughout,NHAC/VEGF group is the second.Conclusion:1,Compare to three kinds of materials, osteoblasts in allogeneic bone grows best, NHAC/VEGF composite bone is the second, and group bone cement is on the worst.2,NHAC/VEGF has good compatibility with cells, which can be a new kind of composite artificial bone grafts.