Enhance adhesion, proliferation and mineralization of osteoblasts cultured on nano-porous surface of titanium alloy

摘要

Objective: To study osteoblast adhesion, proliferation and ossification on nanoporous surface of titanium alloy. Material and method: Scanning electron microscope (SEM) and atomic force microscope (AFM) were used to identify the nanoporous surface on titanium alloy. Osteoblasts from Cranial bone of SD rat were cultured on nanoporous (N) surface titanium alloy and smooth (S) and rough (R) surface titanium alloy were references. The osteoblast adhesion, proliferation and ossification were observed and analyzed by fluorescent microscope (FM), SEM, MTT colorimetric method, Image-Pro Plus (IPP) software respectively. The data were analyzed by SPSS. Result: The osteoblast was stained by AO and counted by IPP. The result showed that the adhered cells increased significantly with time (P<0.05) in all three groups and the number of cells on N group was greater than other two at certain time (P<0.05). The SEM observation showed that osteoblast on the nano-surface spread earlier than other two. The growth curve according A which was determined by MTT colorimetric method showed that the curve of three different groups had similar appearance. But exponential phase of growth of N group was longer than S and R group. And its peak was higher than other two. The doubling generation time of S, R and N was 3.0d, 3.0d, 2.7d.The area of ossified nodules on the three different material also showed that the area of N group was larger than other two (P<0.05). Discussion: Osteoblast cultured on material showed that after the surface had been nano-structured, the osteoblast reached earlier adhesion, more powerful proliferation and ossification. The roughness of material increased a lot through nano-technology, so the osteoblast had much more surface area to contact with it. The other reason may be the change of electrochemistry which led more RGD proteins to cover the material which give more sites to bind with integrin. Conclusion: Nanoporous surface technology can increase the early adhesion, proliferation and ossification of osteoblast in vitro.