Formation of three-dimensional cell/polymer constructs for bone tissue engineering in a spinner flask and a rotating wall vessel bioreactor
Formation of three-dimensional cell/polymer constructs for bone tissue engineering in a spinner flask and a rotating wall vessel bioreactor
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DOI：
10.1002/jbm.10150
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发表时间：
2002-10-01
期刊：
影响因子：
--
通讯作者：
Mikos, AG
中科院分区：
文献类型：
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作者：
Sikavitsas, VI;Bancroft, GN;Mikos, AG
本研究的目的是研究不同生物反应器培养的大鼠骨髓基质细胞（MSCs）对三维聚合物支架细胞增殖、向成骨细胞分化和生成矿化细胞外基质能力的影响。从雄性Sprague-Dawley大鼠身上收获的MSCs进行扩增培养，将其植入三维多孔75:25聚（D， l -乳酸-羟基乙酸）可生物降解支架上，并在静态条件下或在两个模型生物反应器（旋转瓶和旋转壁管）中培养21天，以增强培养基的混合，并为种子细胞提供更好的营养运输。旋转瓶培养显示为60%。与静态培养相比，在第一周结束时增殖增强。在第14天，所有细胞/聚合物结构的碱性磷酸酶活性（AP）均达到最大值。第14天，在旋转烧瓶中培养的细胞/聚合物构建物的AP活性比在静态条件下培养的构建物高2.4倍。体外培养的骨钙素（OC）分泌量是静态培养的3.5倍，在第18天达到峰值。在21天的培养期间，在旋转壁血管培养中未检测到明显的AP活性和OC分泌。旋转瓶培养在第14天钙含量最高。在第21天，旋转烧瓶培养的钙沉积比静态培养的构建物高6.6倍，比旋转壁容器培养高30倍以上。组织学切片显示第21天泡沫表面细胞浓度和矿化。这种现象可能是由于支架内部可能存在营养浓度梯度引起的。在支架外表面外的旋转烧瓶中提供更好的混合，可能解释了骨髓基质成骨细胞增殖和分化的加速，以及支架外表面矿化增强的定位。(C) 2002 Wiley期刊有限公司
The aim of this study is to investigate the effect of the cell culture conditions of three-dimensional polymer scaffolds seeded with rat marrow stromal cells (MSCs) cultured in different bioreactors concerning the ability of these cells to proliferate, differentiate towards the osteoblastic lineage, and generate mineralized extracellular matrix. MSCs harvested from male Sprague-Dawley rats were culture expanded, seeded on three-dimensional porous 75:25 poly(D,L-lactic-co-glycolic acid) biodegradable scaffolds, and cultured for 21 days under static conditions or in two model bioreactors (a spinner flask and a rotating wall vessel) that enhance mixing of the media and provide better nutrient transport to the seeded cells. The spinner flask culture demonstrated a 60%. enhanced proliferation at the end of the first week when compared to static culture. On day 14, all cell/polymer constructs exhibited their maximum alkaline phosphatase activity (AP). Cell/polymer constructs cultured in the spinner flask had 2.4 times higher AP activity than constructs cultured under static conditions on day 14. The total osteocalcin (OC) secretion in the spinner flack culture was 3.5 times higher than the static culture, with a peak OC secretion occurring on day 18. No considerable AP activity and OC secretion were detected in the rotating wall vessel culture throughout the 21-day culture period. The spinner flask culture had the highest calcium content at day 14. On day 21, the Calcium deposition in the spinner flask culture was 6.6 times higher than the static cultured constructs and over 30 times higher than the rotating wall vessel culture. Histological sections showed concentration of cells and mineralization at the exterior of the foams at day 21. This phenomenon may arise from the potential existence of nutrient concentration gradients at the interior of the scaffolds. The better mixing provided in the spinner flask, external to the outer surface of the scaffolds, may explain the accelerated proliferation and differentiation of marrow stromal osteoblasts, and the localization of the enhanced mineralization on the external surface of the scaffolds. (C) 2002 Wiley Periodicals, Inc.