氯磷酸二钠表面改性羟基磷灰石对骨髓间充质干细胞多向分化的影响

Effect of Clodronate Modifying Hydroxyapatite on Mesenchymal Stem Cells Multi-directional Differentiation

目的观察羟基磷灰石经氯磷酸二钠表面改性后对骨髓间充质干细胞多向分化的影响。方法将块状羟基磷灰石与氯磷酸二钠复合制成氯磷酸二钠-羟基磷灰石,作为实验组;未复合氯磷酸二钠的羟基磷灰石作为对照组。分离,纯化大鼠的骨髓间充质干细胞,培养、传代,应用免疫细胞化学方法对培养至第3代骨髓间充质干细胞进行鉴定。将两组标本与骨髓间充质干细胞复合培养,利用MTT法对比骨髓间充质干细胞在两组材料表面生长有无差异,并且分别进行骨向、肌向、脂向诱导,观察氯磷酸二钠-羟基磷灰石对骨髓间充质干细胞多向分化有无影响。结果表面标志物SH3的免疫细胞化学检测证明本实验获得的第3代骨髓间充质干细胞具有原始干细胞的特性。利用MTT法吸光度分析检测第3代骨髓间充质干细胞在两组标本的生长曲线,经统计学检验P>0.05,两组之间无统计学差异。骨髓间充质干细胞经骨向、肌向、脂向诱导后,仍保持成骨、成肌、成脂能力。结论氯磷酸二钠复合在羟基磷灰石表面对骨髓间充质干细胞多向分化功能没有显著影响。

Objective To investigate the effect of clodronate modifying hydroxyapatite on mesenchymal stem cells （BMSCs） multi - directional differentiation. Methods Samples in the experimental group were combined with the clodronate to form compound materials, while samples in the control group were hydroxyapatite without clodronate. BMSCs of rats were cultured, and then identified by immunocytochemical method. Samples in two groups were co - cultured with BMSCs. Then MTT method and multi - directional induction were used to investigate the effect of clodronate modifying hydroxyapatite on BMSCs differentiation. Results Immunocytochemical method about surface marker SH3 demonstrated that BMSCs of the third generation still kept the characteristic of primitive stem cells. MTT absorbancc method detected the growth curve of the third generation BMSCs on sample surface in both groups,and there was no significant difference between the two groups（P 〉0.05）. With induction of multi -direction differentiation,BMSCs still kept the multi- directional differentiation capability. Conclusion Clodronate modified hydroxyapatite has no significant effect on multi - directional differentiation capability of BMSCs which were located on material surface.