Stoker Aaron, Garrity Joeseph T, Hung Clark T, Stannard James P, Cook Jimi
Department of Veterinary Medicine and Surgery, University of Missouri, Columbia, Missouri 65211, USA.
J Knee Surg. 2012 May;25(2):117-25. doi: 10.1055/s-0032-1319809.
Fresh osteochondral allografts (OCAs) have been used clinically to treat cartilage focal defects of the knee for over 30 years. Over the last decade, significant research has been performed to develop and improve protocols for preservation of osteochondral tissue before transplantation into patients for treatment of cartilage defects. This work has resulted in preservation protocols that allow for maintenance of OCA tissues for time periods sufficient for clinical use based on disease testing requirements in the United States. However, graft quality and the window for clinical use of these tissues could be greatly enhanced from current levels.
Femoral condyles from 14 dogs were harvested and stored in one of the three proprietary media composition (M-1, M-2, M-3) and container condition (C-1, C-2, C-3) at 25 degrees C for 63 days. Viability of the OCA was determined using a proprietary media metabolic assay and live cell fluorescent microscopy. Media biomarker concentrations were analyzed to determine the metabolic activity of tissue.
Media protein biomarkers were detected throughout the culture period, indicating OCAs remain metabolically active at 25 degrees C, and biomarker levels correlated with tissue viability. Viable chondrocyte density was maintained at day 0 levels throughout the depth of the tissue in the M-3 media using container condition C-3 after 63 days in storage. The media metabolic assay correlated strongly to cell viability of the OCA tissue.
These data indicate that near day 0 tissue viability can be maintained for up to 63 days when OCAs are stored at 25 degrees C in the correct conditions. Further, tissue viability could be assessed nondestructively using media biomarkers and the media metabolic assay. If the preservation protocol reported here can be validated for safety and functional outcome, it could then be employed in tissue banks throughout the world, decreasing the number of grafts discarded and improving quality of life for thousands of patients affected by cartilage defects.
新鲜骨软骨异体移植（OCA）已在临床上用于治疗膝关节软骨局灶性缺损超过30年。在过去十年中，已经开展了大量研究，以开发和改进在将骨软骨组织移植到患者体内治疗软骨缺损之前的保存方案。这项工作产生了一些保存方案，这些方案能够根据美国的疾病检测要求，将OCA组织维持足够长的时间以供临床使用。然而，这些组织的移植质量和临床使用期限仍可从当前水平大幅提高。
从14只犬获取股骨髁，并在25摄氏度下，将其储存在三种专利培养基成分（M-1、M-2、M-3）和容器条件（C-1、C-2、C-3）之一中，保存63天。使用专利培养基代谢测定法和活细胞荧光显微镜检查来确定OCA的活力。分析培养基生物标志物浓度以确定组织的代谢活性。
在整个培养期间都检测到了培养基蛋白生物标志物，表明OCA在25摄氏度下仍保持代谢活性，并且生物标志物水平与组织活力相关。在储存63天后，使用容器条件C-3的M-3培养基中，整个组织深度的活软骨细胞密度在第0天的水平维持不变。培养基代谢测定与OCA组织的细胞活力密切相关。
这些数据表明，当OCA在正确条件下于25摄氏度储存时，接近第0天的组织活力可维持长达63天。此外，可以使用培养基生物标志物和培养基代谢测定法无损评估组织活力。如果此处报告的保存方案能够在安全性和功能结果方面得到验证，那么它就可以在世界各地的组织库中使用，减少废弃移植物的数量，并改善数千名受软骨缺损影响患者的生活质量。
