Akmal M, Anand A, Anand B, Wiseman M, Goodship A E, Bentley G
Institute of Orthopaedics, Stanmore, Middlesex, UK.
J Bone Joint Surg Br. 2006 Apr;88(4):544-53. doi: 10.1302/0301-620X.88B4.16498.
Bovine and human articular chondrocytes were seeded in 2% alginate constructs and cultured for up to 19 days in a rotating-wall-vessel (RWV) and under static conditions. Culture within the RWV enhanced DNA levels for bovine chondrocyte-seeded constructs when compared with static conditions but did not produce enhancement for human cells. There was a significant enhancement of glycosaminoglycans and hydroxyproline synthesis for both bovine and human chondrocytes. In all cases, histological analysis revealed enhanced Safranin-O staining in the peripheral regions of the constructs compared with the central region. There was an overall increase in staining intensity after culture within the RWV compared with static conditions. Type-II collagen was produced by both bovine and human chondrocytes in the peripheral and central regions of the constructs and the staining intensity was enhanced by culture within the RWV. A capsule of flattened cells containing type-I collagen developed around the constructs maintained under static conditions when seeded with either bovine or human chondrocytes, but not when cultured within the RWV bioreactor.
将牛和人的关节软骨细胞接种到2%的藻酸盐构建物中,在旋转壁式生物反应器(RWV)中以及静态条件下培养长达19天。与静态条件相比,RWV内的培养提高了接种牛软骨细胞构建物的DNA水平,但对人细胞没有产生增强作用。牛和人软骨细胞的糖胺聚糖和羟脯氨酸合成均有显著增强。在所有情况下,组织学分析显示,与构建物中央区域相比,其外周区域的番红O染色增强。与静态条件相比,在RWV内培养后染色强度总体增加。构建物外周和中央区域的牛和人软骨细胞均产生II型胶原蛋白,且在RWV内培养可增强染色强度。当接种牛或人软骨细胞时,在静态条件下培养的构建物周围会形成一层含有I型胶原蛋白的扁平细胞囊,但在RWV生物反应器内培养时则不会。