Caruso E M, Lewandrowski K U, Ohlendorf C, Tomford W W, Zaleske D J
Orthopaedic Research Laboratories, Massachusetts General Hospital, Boston, 02114, USA.
J Orthop Res. 1996 Jan;14(1):102-7. doi: 10.1002/jor.1100140117.
Growth of chondrocytes into a xenogeneic chondroepiphyseal matrix was investigated in an in vitro experimental model by combining viable calf chondrocytes with chick epiphyseal matrix devoid of viable chondrocytes. The chondrocytes were harvested from the wrist joints of newborn calves and cultured for 2 days. The epiphyses were harvested from the distal femurs and the proximal tibias of fetal chicks after development was arrested at 17 days by freezing. The epiphyseal specimens were prepared in four ways. These included femoral and tibial epiphyses without holes and femoral and tibial epiphyses with holes made by a laser. These epiphyseal specimens were co-cultured with calf chondrocytes for various periods. After digestion of the epiphyseal matrix, viable chondrocytes were counted in suspension. Chondrocyte division in the matrix was assessed by [3H]thymidine incorporation. The growth of calf chondrocytes into the xenogeneic chick matrix was evaluated by fluorescence microscopy on fresh thick epiphyseal sections. The percentage of viable chondrocytes in the xenogeneic epiphyseal matrix increased with culture time to a maximum at day 21. The addition of laser-drilled holes was found to extend a plateau of chondrocyte viability until day 29. A decrease in cell viability was detected at later observation points. This study demonstrates that xenogeneic matrix may serve as a morphogenetic scaffold for chondrocytic growth.
通过将存活的小牛软骨细胞与不含存活软骨细胞的鸡骨骺基质相结合,在体外实验模型中研究了软骨细胞在异种骨骺基质中的生长情况。软骨细胞从小牛新生犊牛的腕关节获取,并培养2天。在发育至17天时通过冷冻使其停止发育后,从胎鸡的股骨远端和胫骨近端获取骨骺。骨骺标本通过四种方式制备。这些方式包括无孔的股骨和胫骨骨骺以及有激光打孔的股骨和胫骨骨骺。将这些骨骺标本与小牛软骨细胞共培养不同时间段。在消化骨骺基质后,对悬浮液中的存活软骨细胞进行计数。通过[3H]胸腺嘧啶核苷掺入评估基质中的软骨细胞分裂。通过对新鲜厚骨骺切片进行荧光显微镜检查来评估小牛软骨细胞在异种鸡基质中的生长情况。异种骨骺基质中存活软骨细胞的百分比随培养时间增加,在第21天达到最大值。发现添加激光打孔可将软骨细胞活力的平稳期延长至第29天。在后期观察点检测到细胞活力下降。本研究表明,异种基质可作为软骨细胞生长的形态发生支架。