Department of Bioengineering, University of California-San Diego, 9500 Gilman Drive, Mail Code 0412, La Jolla, CA 92093-0412, USA.
Tissue Eng Part A. 2010 May;16(5):1717-27. doi: 10.1089/ten.TEA.2009.0663.
Understanding and controlling chondrocyte and cartilage metabolism in osteochondral tissues may facilitate ex vivo maintenance and application, both for allografts and tissue-engineered grafts. The hypothesis of this study was that maintenance of chondrocyte viability and matrix content and release of sulfated glycosaminoglycan (sGAG) in the articular cartilage of joint-scale osteochondral fragments are temperature and metabolism dependent. The aims were to assess, for adult goat joints, the effects of incubation temperature (37 degrees C vs. 4 degrees C) on cartilage chondrocyte viability and tissue matrix content and mechanical function, and the effects of temperature and cellular biosynthesis on sGAG release. Chondrocyte viability was maintained with 37 degrees C incubation for 28 days, but decreased by approximately 30% with 4 degrees C incubation. Concomitantly, with 37 degrees C incubation, cartilage sGAG was depleted by approximately 52% with the lost sGAG predominantly unable to aggregate with hyaluronan, whereas collagen content, tissue thickness, and tissue stiffness were maintained. The depletion of sGAG was diminished by slowing metabolism, with 4 degrees C decreasing release by approximately 79% compared with 37 degrees C incubation, and cycloheximide inhibition of cell metabolism at 37 degrees C decreasing release by approximately 47%. These results indicate that the articular cartilage of joint-scale grafts have enhanced chondrocyte viability with incubation at 37 degrees C, but may need anabolic stimuli or catabolic inhibitors to maintain sGAG content.
了解和控制软骨细胞和软骨代谢在骨软骨组织中,可以促进同种异体移植物和组织工程移植物的体外维持和应用。本研究的假设是,维持关节尺度骨软骨片段关节软骨中的软骨细胞活力和基质含量以及硫酸化糖胺聚糖(sGAG)的释放与温度和代谢有关。目的是评估成年山羊关节,孵育温度(37°C 与 4°C)对软骨细胞活力和组织基质含量和机械功能的影响,以及温度和细胞生物合成对 sGAG 释放的影响。37°C 孵育 28 天可维持软骨细胞活力,但 4°C 孵育时活力降低约 30%。同时,在 37°C 孵育时,软骨 sGAG 耗竭约 52%,丢失的 sGAG 主要不能与透明质酸聚集,而胶原含量、组织厚度和组织硬度保持不变。降低代谢速度可以减少 sGAG 的耗竭,4°C 孵育时的释放量比 37°C 孵育时减少约 79%,而 37°C 时细胞代谢的环己亚胺抑制作用使释放量减少约 47%。这些结果表明,关节尺度移植物的关节软骨在 37°C 孵育时具有增强的软骨细胞活力,但可能需要合成代谢刺激或分解代谢抑制剂来维持 sGAG 含量。