骨髓间充质干细胞膜片复合磷酸三钙陶瓷构建工程化骨组织的体内成骨研究(1)
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[摘要]目的：探讨骨髓间充质干细胞膜片复合磷酸三钙支架材料构建组织工程骨的可行性。方法：将兔骨髓间充质干细胞高密度接种于普通培养皿，在成骨诱导条件下连续培养2周，获得细胞膜片，修剪成长方形，并由一端卷起包裹圆柱状的磷酸三钙材料。静置孵育24h后将构建物移植到裸鼠背部皮下。术后6周取材，进行大体观察、组织学检查、组织定量学分析。结果：所获骨髓间充质干细胞膜片有多层细胞组成，保留了细胞外基质。实验组在材料表面及其孔隙内有较多的骨质形成；单一材料组空隙内为纤维组织，未见骨或软骨样组织；单一膜片组见片状编织骨形成。 结论：骨髓间充质干细胞膜片在体内具有良好的成骨能力，可作为细胞释放载体与磷酸三钙支架复合构建骨组织。本研究为骨组织工程构建提供了新的方法。
[关键词]骨髓间充质干细胞；骨组织工程；细胞膜片；磷酸三钙
[中图分类号]R318[文献标识码]A[文章编号]1008-6455(2011)03-0408-03
Engineering bone tissue using bone marrow mesenchymal stem cell sheet and β-tricalcium phosphate ceramic
MA Dong-yang1,MA Jing2,JIANG Dong-hong3,WANG Jian-feng3
(1.Department of Oral and Maxillofacial Surgery,2. Department of Otolaryngology,3. Department of Medical Administration，Lanzhou General Hospital,Lanzhou Command,PLA,730052,Gansu,China)
Abstract:ObjectiveTo investigate the feasibility of constructing bone tissue using bone marrow mesenchymal stem cell (BMSC) sheet and β-tricalcium phosphate ceramic (TCP).MethodsWe first harvested a cell sheet from rabbit BMSCs using a continuous culture method and a scraping technique. The cell sheet was then wraped around a cylinder of β-TCP. Finally,the constructs were implanted into the subcutaneous pockets of nude mice for in vivo experiments. Gross view and histological examinations were performed to evaluate the harvested specimens. Results The cell sheet, with an average thickness of 158 mm, was composed of multi-layered cells separated in the extracellular matrix. Six weeks after implantation, the new bone tissue was present both on the edge and at the center of the TCP in sheet-TCP group. A layer of woven bone formed in the cell-sheet group. In contrast, the TCP was filled only with fibrous tissue in the TCP group,without evidence of bone formation.ConclusionThe study indicates that the combination of osteogenic BMSC sheet and β-TCP ceramic can engineer bone tissue and the engineered construct might be considered as a promising substitute for bone repair.
Key words:bone mesenchymal stem cell; bone tissue engineering; cell sheet; β-tricalcium phosphate
近年来，随着再生医学的迅速发展，组织工程有望为骨缺损提供理想的修复方法[1]。组织工程骨的传统构建方法是将具有成骨分化功能的种子细胞与三维支架材料复合[1-2]。细胞在体外扩增后常用胰蛋白酶消化成离散细胞悬液，在此过程中，细胞自分泌的细胞外基质、细胞-基质连接、以及细胞-细胞连接等结构都被破坏，而这些结构对于细胞分化、特异性表型维持、分泌、组织形成等功能非常重要[3] ......
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