人工颈椎间盘不同涂层影响骨髓间充质干细胞的黏附及分化

摘要

BACKGROUND:Titanium artificial cervical disc has good biocompatibility, but titanium al oy is shown to have poor biological activity, low bonding strength, easy release of metal ions under physiological conditions. OBJECTIVE:To observe the effects of different coating for the titanium plate of domestic cervical artificial disc on adhesion and differentiation ability of rat bone marrow mesenchymal stem cel s. METHODS:Passage 3 bone marrow mesenchymal stem cel s from Wistar rats were seeded into 24-wel titanium plates with hydroxyapatite coating, titanium powder+hydroxyapatite coating and bare titanium plate. Cel culture was terminated after 24 and 48 hours, and the cel growth was observed under scanning electron microscope. After 24 hours of inoculation, osteogenic inducer was added;then, cel supernatant was col ected at 3, 5, 7 days after cel lysis and centrifugation to detect the activity of alkaline phosphatase. RESULTS AND CONCLUSION:After composite culture with titanium plates with hydroxyapatite coating or titanium powder+hydroxyapatite coating, cultured cel s were in polygonal shape, and pseudopodia were extended into the micropores that were adhered closely to the material surface. Cel s cultured with bare titanium plates had poor differentiation and adhesion rate. With time, the expression of alkaline phosphatase was increased in each group, especial y in the groups of titanium plates with hydroxyapatite coating and titanium powder+hydroxyapatite coating (P<0.05). These findings indicate that titanium plates with hydroxyapatite coating or titanium powder+hydroxyapatite coating can promote adhesion and differentiation of rat bone marrow mesenchymal stem cel s.%背景：钛合金人工颈椎间盘具有良好的生物相容性，但钛合金存在生物活性差、与骨结合强度低、在生理环境下易造成金属离子释放等问题。　　目的：观察国产人工颈椎间盘钛合金终板不同涂层对大鼠骨髓间充质干细胞黏附、分化的影响。　　方法：将第3代Wistar大鼠骨髓间充质干细胞，接种于含羟基磷灰石涂层、钛粉复合羟基磷灰石涂层的钛合金板及裸钛合金板的24孔板内，培养的第24，48小时后分别终止培养，扫描电镜观察细胞生长情况；接种24 h后加成骨诱导剂进行诱导，并于第3，5，7天各收集细胞裂解后离心的上清，检测碱性磷酸酶表达。结果与结论：骨髓间充质干细胞与材料复合培养48 h后，羟基磷灰石组和钛粉复合羟基磷灰石组表面的细胞呈多角形，并伸出细长伪足伸入到材料的微孔内，与材料表面紧密黏附；而裸钛合金组骨髓间充质干细胞分化较差且黏附率低。随时间延长，各组碱性磷酸酶表达均增加，羟基磷灰石组和钛粉复合羟基磷灰石组各时间点的碱性磷酸酶表达均较裸钛合金组明显增高(P<0.05)。表明羟基磷灰石、钛粉复合羟基磷灰石涂层可有促进大鼠骨髓间充质干细胞的黏附和分化。