新型多孔磷酸钙支架对骨髓间充质干细胞生物学行为的影响
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[摘要] 目的 评价新型多孔磷酸钙（CPC）作为骨组织工程支架对Beagle犬骨髓间充质干细胞（BMSCs）黏附、增殖及成骨分化等生物学行为的影响。方法 将Beagle犬BMSCs接种于新型多孔CPC三维支架表面，以磷酸三钙（TCP）和聚乳酸-聚羟基乙酸共聚物（PLGA）为对照组，通过观察细胞形态、绘制细胞生长曲线、测定碱性磷酸酶（ALP）活性、进行茜素红染色并半定量测定骨钙素等方法，检测BMSCs在支架材料上的黏附、增殖及成骨分化情况。结果 细胞形态和生长曲线结果显示BMSCs在新型多孔CPC三维支架材料表面分布均匀，生长及增殖活跃。ALP活性半定量结果表明，CPC、TCP组ALP表达强度明显高于PLGA组（P＜0.05），CPC组与TCP组间的差异无统计学意义（P＞0.05）。骨钙素染色与半定量检测结果均显示：各观察点PLGA组钙盐沉积量明显少于CPC和TCP组（P＜0.05），而TCP和CPC
组间的差异无统计学意义（P＞0.05）。结论 本实验所用多孔CPC材料具有与TCP类似但优于PLGA的良好生物相容性，
利于BMSCs的黏附、增殖及成骨分化，可作为支架材料与BMSCs共同培养，构建具有成骨能力的组织工程化骨。
[关键词] 多孔磷酸钙； 骨髓间充质干细胞； 黏附； 增殖； 分化
[中图分类号] R 78 [文献标志码] A [doi] 10.3969/j.issn.1000-1182.2012.05.022
Influence of novel porous calcium phosphate cement on biological behavior of bone marrow mesenchymal stem cells Wang Shuhong1，2, Zhang Xiong2, Zhang Jin’e1, Huang Yuanliang1，3. （1. School of Stomatology, Tongji Uni-versity, Shanghai 200072, China; 2. Dept. of Stomatology, Hangzhou First People’s Hospital, Hangzhou 310006, China; 3. Dept. of Stomatology, Shanghai East Hospital Affiliated to Tongji University, Shanghai 200120, China）
[Abstract] Objective To investigate the effect of novel porous calcium phosphate cement（CPC） scaffoldings on
attachment, proliferation and differentiation of bone marrow mesenchymal stem cells（BMSCs）. Methods BMSCs of
Beagle dogs were implanted and cultured with CPC scaffoldings in vitro, tricalcium phosphate（TCP） and poly（lactide-co-glycolide） （PLGA） scaffoldings as controls. The attachment, proliferation and differentiation of BMSCs were detected
through morphological characters, growth curve and the semi-quantitative detection of alkaline phosphatase（ALP） and osteocalcin（OC） activity. Results Cell morphology and growth curve illustrated that BMSCs attached to and grown
better on the surface of novel porous CPC scaffoldings than that of PLGA group（P
of ALP showed that ALP expression level in BMSCs on the CPC and TCP group were significantly higher than that of the PLGA group（P＜0.05）, the CPC group was slightly higher than the TCP group, but no significant difference
was found between the two groups（P＞0.05）. The staining and semi-quantitative analysis results of OC demonstrated
that calcium deposition of the PLGA group was significantly less than the CPC and TCP group on both observation point（P＜0.05）, but no significant difference between the CPC and TCP group（P＞0.05）. Conclusion The novel porous
CPC material used in this study has good biocompatibility similar to TCP but much better than PLGA which is fa-vorable of BMSCs adhesion, proliferation and osteogenic differentiation. The novel porous CPC material is a suitable scaffolding for BMSCs to fabricate tissue-engineered bone in vitro.