钛表面微弧氧化涂层的细胞生物活性

背景：微弧氧化技术可改善钛或钛合金的表面特征。目的：研究纯钛表面微弧氧化涂层的表面性能及其对MC3T3-E1细胞早期黏附、增殖及成骨能力的影响。方法：将46个直径10 mm、厚度2 mm圆盘状纯钛试件分为实验组和对照组。实验组置于含0.02 mol/Lβ-甘油磷酸二钠盐及0.2 mol/L乙酸钙的电解液中进行微弧氧化处理,对照组对试件进行机械抛光。扫描电子显微镜观察试件表面形貌,X射线能谱分析检测涂层表面钙磷比,X射线衍射分析检测涂层晶相构成。将MC3T3-E1细胞接种在两组试件表面,1,2,4 h电镜下观察细胞形态,在2,4,7 d通过CCK-8方法检测细胞增殖,并于7,14 d检测碱性磷酸酶活性。结果与结论：经微弧氧化处理后,钛表面形成粗糙多孔的钙磷涂层,微弧氧化涂层主要元素为Ca、P、O及Ti,微弧氧化膜层主要由氧化钛、钛酸钙、磷酸钙及偏磷酸钙构成。电镜观察显示1 h 微弧氧化涂层表面细胞已伸出伪足,4 h呈现较典型的细胞形态。细胞在微弧氧化处理钛表面4,7 d的细胞增殖和7,14 d的碱性磷酸酶活性高于对照组。表明微弧氧化技术生成的粗糙多孔钙磷涂层能显著促进MC3T3-E1细胞的早期黏附、增殖及成骨活性。

Cell bioactivity of micro-arc oxidation coating on the surface of pure titanium

BACKGROUND:Micro-arc oxidation technique is used to modify the surface properties of titanium and titanium alloy.  OBJECTIVE:To explore the surface properties of micro-arc oxidation film and its effect on the attachment, proliferation and alkaline phosphatase activity of MC3T3-E1. METHODS:Forty-six pure titanium discs, 10 mm in diameter and 2 mm in thickness, were randomly divided into two groups. The discs of the experimental groups were treated by micro-arc oxidation technique in an electrolytic solution containing 0.02 mol/L sodium β-glycerophosphate and 0.2 mol/L calcium acetate; while the discs of the control group was machine-polished. The surface appearance of the discs was observed by a scanning electron microscopy, the ratio of calcium to phosphorus on the coating surface was detected by X-ray spectroscopy, and the crystalline phase composition of the coating was detected by X-ray diffraction analysis. Cellular morphology in the process of attachment was observed under the scanning electron microscope. Cell proliferation was determined by cell counting kit-8 at 2, 4, 74 days, while alkaline phosphatase activity were determined at 7 and 14 days. RESULTS AND CONCLUSION:After micro-arc oxidation treatment, a rough and porous calcium-phosphate film was formed on the surface of titanium. The elements of micro-arc oxidation coating main mainly included Ca, P, O and Ti, and the micro-arc oxidation film was mainly composed of titanium oxide, calcium titanate, calcium phosphate and calcium metaphosphate. Under the scanning electron microscope, pseudopods appeared to grow out of the cells on the surface of micro-arc oxidation coating after 1 hour culture, and the typical morphology of the MC3T3-E1 cells could be observed at 4 hours. MC3T3-E1 proliferation (4 and 7 days of culture) and alkaline phosphatase activity (7 and 14 days of culture) were enhanced significantly in the micro-arc oxidation group compared with the control group. These findings indicate that the rough and porous calcium-phosphate coating produced by micro-arc oxidation technique on pure titanium surface could promote the early attachment, proliferation and osteogenic differentiation of MC3T3-E1.