利用灌注式生物反应器体外构建大段组织工程化骨的实验研究  

Construction of large-scale tissue-engineered bone using a perfusion bioreactor

作　　者：李德强[1] 戴尅戎[1] 汤亭亭[1] 卢建熙 

机构地区：[1]上海交通大学医学院附属第九人民医院骨科,200011 [2]上海贝奥路生物材料有限公司

出　　处：《中华创伤骨科杂志》2009年第5期455-459,共5页Chinese Journal of Orthopaedic Trauma

基　　金：国际科技合作重点项目（2005DFA30120）;国家重点基础研究发展计划（973项目）（2005cB522700）;国家自然科学基金（30600629）;高等学校博士学科点专项科研基金（200802480070）;上海交通大学医学院博士创新基金（BXJ0821）

摘　　要：目的研究体外利用灌注式生物反应器构建大段组织工程化骨的可行性。方法把在体外培养扩增的第三代人骨髓基质干细胞与大段多孔β-磷酸三钙（β-TCP）支架复合。将细胞／支架复合体放入灌注式生物反应器中，进行连续灌注培养。28d后，检测细胞的增殖及碱性磷酸酶（ALP）活性，同时对培养后的细胞／支架复合体进行组织学检测及形态学计量，用以评价体外组织工程化骨的构建。以静态培养作为对照组。结果培养28d后，灌注培养组的细胞活性明显高于静态培养组。灌注培养组细胞的ALP活性显著高于静态培养组。静态培养组细胞仅在多孔β-TCP支架周缘增殖，形成的新骨量较少。灌注培养组细胞在整个β—TCP支架内增殖，形成的新骨量较多。结论利用灌注式生物反应器的灌注培养，可以使人骨髓基质干细胞在大段β-TCP载体内增殖并形成新骨，使体外大段组织工程化骨的构建成为可能。Objective To study the feasibility of constructing a large-scale tissue-engineered bone using a perfusion bioreactor. Methods Human bone marrow stromal cells （BMSCs） were isolated from human bone marrow and cultured. After passage and proliferation, the human BMSCs were seeded onto the critical-size porous β-tficalcium phosphate （β-TCP） scaffold and cultured in a perfusion bioreactor. After 28 days of culture, the construction of a large-scale tissue-engineered bone was assessed by the proliferation and osteogenic differentiation of cells and histological assay and histomorphometry of the cells β-TCP construct. The static culture served as a control. Results After 28 days of culture, the cell viability was higher in perfusion culture than in static culture. And the alkaline phosphatase activity was also higher in perfusion culture than in static culture. In static culture, human BMSCs only survived and proliferated in the periphery of the β-TCP scaffold, and the new bone volume was small. However, in perfusion culture, the cells survived and proliferated in the whole scaffold, and the new bone volume was larger. Conclusions After perfusion culture in a perfusion bioreactor, the human BMSCs can survive, proliferate and form new bone tissue in the critical-size porous scaffold. As a result, a large-scale tissue-engineered bone can be constructed in vitro using a perfusion bioreactor.

关 键 词：生物反应器 组织工程 骨髓细胞 β-磷酸三钙支架 

分 类 号：R686[医药卫生—骨科学]