灌注式生物反应器中流体剪切力对大段组织工程化骨构建的作用  

Study on the effects of flow shear stress in constructing large-scale tissue-engineered bone using a perfusion bioreactor

作　　者：李德强[1,5,6] 杨爱玲[2] 汤亭亭[1,5] 卢建熙[1,4] 郭雪岩[2] 戴魁戎[1,3,5,6] 

机构地区：[1]上海交通大学医学院附属第九人民医院骨科,上海200011 [2]上海理工大学动力工程学院,上海200093 [3]上海生命科学研究院上海交通大学医学院健康科学研究所骨科细胞分子生物学实验室,上海200025 [4]上海贝奥路生物材料有限公司,上海200335 [5]上海市骨科内植物重点实验室,上海200011 [6]教育部数字医学工程研究中心,上海200030

出　　处：《医用生物力学》2009年第1期8-14,20,共8页Journal of Medical Biomechanics

基　　金：国际科技合作重点项目(2005DFA30120);国家自然科学基金资助项目(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);国家自然科学基金资助项目(30600628)

摘　　要：目的结合流体力学模型研究灌注式生物反应器中大段组织工程化骨的构建与多孔支架内流体剪切力的关系。方法利用灌注式生物反应器对复合骨髓基质干细胞的多孔磷酸三钙支架进行灌注培养。培养基的黏度分别为1.12mPa.s,2.23mPa.s及3.35mPa.s。通过细胞增殖、成骨分化及组织形态学评价组织工程化骨的构建,建立流体力学模型,求解支架内的流体剪切力。结果培养基黏度2.23mPa.s组,细胞增殖高于其他组。培养基黏度2.23mPa.s及3.35mPa.s组第28d的碱性磷酸酶活性及第7d后的骨钙素分泌高于1.12mPa.s组。培养基黏度越高,骨桥蛋白的分泌高峰出现越早。28d后,黏度3.35mPa.s组的钙化基质最多。流体力学模型分析,培养基黏度1.12mPa.s,2.23mPa.s及3.35mPa.s组中,支架内的平均流体剪切力分别为5mPa,11mPa和15mPa。结论在利用复合人骨髓基质干细胞的多孔磷酸三钙构建大段组织工程化骨的过程中,15mPa的流体剪切力最有利于组织工程化骨的构建。Objective To study the relationship between the construction of large engineered bone and flow shear stress inside the β-tricalcium phosphate (β-TCP) scaffold in perfusion bioreactor with the flow model. Method The scaffolds seeded with human bone marrow-derived mesenchymal stem cells were cultured in perfusion bioreactor. The viscosity of the media was 1.12 mPa·s ,223 mPa·s, and 335 mPa·s respectively. The construclion of the tissue-engineered bone was assessed through the proliferation and osteogenic differentiation of cells and histological assay of the constructs. The flow model was established to calculate the flow shear stress inside the β-TCP scaffold. Result The cell viability was highest in the group with the viscosityof223 mPa·s. The alkaline phosphatase activity at day28 and the osteocalcin secretion alter 7 days were higher in the two groups with the viscosities of 223 mPa · s and 335 mPa· s than in the group with the with the viscosity of 1.12 mPa ·s. Increasing the viscosity of the medium resulted in an earlier emergence of secretion peak of osteopontin. After 28 days of culture, the mineralized exacellular matrix formed inside the scaffold was most in the group with the viscosity of 3.35 mPa · s. The averaged flow shear stress in the scaffold was 5 mPa, 11 mPa, and 15 Pa accordingly when the viscosity of the media was 1.12 mPa ·s ,223 mPa · s, and 335 mPa ·s respectively. Conclusions In this study, a 15 mPa of flow shear stress was optimal in the construction of a large-scale tissue-engineered bone using the ^-TCP scaffold combined with human bone marrow-derived mesenchymal stem cells in a perfusion bioreactor.

关 键 词：生物反应器 骨组织工程 流体剪切力 计算流体动力学 骨髓基质于细胞 

分 类 号：R318.01[医药卫生—生物医学工程]