CAJ | 학술논문

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.28 d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28 d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.
목적 연구체외이용관주식생물반응기구건대단조직공정화골적가행성. 방법파재체외배양확증적제삼대인골수기질간세포여대단다공β-린산삼개(β-TCP)지가복합.장세포/지가복합체방입관주식생물반응기중,진행련속관주배양.28 d후,검측세포적증식급감성린산매(ALP)활성,동시대배양후적세포/지가복합체진행조직학검측급형태학계량,용이평개체외조직공정화골적구건.이정태배양작위대조조. 결과배양28 d후,관주배양조적세포활성명현고우정태배양조.관주배양조세포적ALP활성현저고우정태배양조.정태배양조세포부재다공β-TCP지가주연증식,형성적신골량교소.관주배양조세포재정개β-TCP지가내증식,형성적신골량교다. 결론이용관주식생물반응기적관주배양,가이사인골수기질간세포재대단β-TCP재체내증식병형성신골,사체외대단조직공정화골적구건성위가능.
Objective To study the feasibility of constructing a large-scale tissue-engineered bone using a perfusion bioreactor. Methods Human hone 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-tricalcium 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 peffusion 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.