软骨细胞在 37°C 下长时间储存的存活率高于在 4°C 下储存的存活率，适用于骨软骨移植物。
Chondrocyte viability is higher after prolonged storage at 37 degrees C than at 4 degrees C for osteochondral grafts.
机构信息
Department of Bioengineering and Whitaker Institute of Biomedical Engineering, University of California-San Diego, 9500 Gilman Drive, La Jolla, CA 92093-0412, USA.
出版信息
Am J Sports Med. 2009 Nov;37 Suppl 1(Suppl 1):24S-32S. doi: 10.1177/0363546509351496. Epub 2009 Oct 27.
BACKGROUND
Osteochondral allografts are currently stored at 4 degrees C for 2 to 6 weeks before implantation. At 4 degrees C, chondrocyte viability, especially in the superficial zone, deteriorates starting at 2 weeks. Alternative storage conditions could maintain chondrocyte viability beyond 2 weeks, and thereby facilitate increased graft availability and enhanced graft quality.
PURPOSE
The objective of the study was to determine the effects of prolonged 37 degrees C storage compared with traditional 4 degrees C storage on chondrocyte viability and cartilage matrix content.
STUDY DESIGN
Controlled laboratory study.
METHODS
Osteochondral samples from humeral heads of adult goats were analyzed (i) fresh, or after storage in medium for (ii) 14 days at 4 degrees C including 10% fetal bovine serum, (iii) 28 days at 4 degrees C including 10% fetal bovine serum, (iv) 28 days at 37 degrees C without fetal bovine serum, (v) 28 days at 37 degrees C including 2% fetal bovine serum, or (vi) 28 days at 37 degrees C including 10% fetal bovine serum. Portions of samples were analyzed by microscopy after LIVE/DEAD staining to determine chondrocyte viability and density, both en face (to visualize the articular surface) and vertically (overall and in superficial, middle, and deep zones). The remaining cartilage was analyzed for sulfated glycosaminoglycan and collagen.
RESULTS
The 37 degrees C storage maintained high chondrocyte viability compared with 4 degrees C storage. Viability of samples after 28 days at 37 degrees C was approximately 80% at the cartilage surface en face, approximately 65% in the superficial zone, and approximately 70% in the middle zone, which was much higher than approximately 45%, approximately 20%, and approximately 35%, respectively, in 4 degrees C samples after 28 days, and slightly decreased from approximately 100%, approximately 85%, and approximately 95%, respectively, in fresh controls. Cartilage thickness, glycosaminoglycan content, and collagen content were maintained for 37 degrees C and 4 degrees C samples compared with fresh controls.
CONCLUSION
The 37 degrees C storage of osteochondral grafts supports long-term chondrocyte viability, especially at the vulnerable surface and superficial zone of cartilage.
CLINICAL RELEVANCE
Storage of allografts at a physiologic temperature of 37 degrees C may prolong storage duration, improve graft availability, and improve treatment outcomes.
背景
目前，同种异体骨软骨在植入前通常在 4°C 下储存 2 至 6 周。在 4°C 下，软骨细胞的活力，尤其是在浅层区域，从第 2 周开始恶化。替代储存条件可以使软骨细胞活力保持在 2 周以上，从而增加移植物的可用性并提高移植物的质量。
目的
本研究的目的是确定与传统的 4°C 储存相比，延长 37°C 储存对软骨细胞活力和软骨基质含量的影响。
研究设计
对照实验室研究。
方法
分析取自成年山羊肱骨头的骨软骨样本：（i）新鲜样本，或（ii）在 4°C 下储存 14 天，其中包含 10%胎牛血清，（iii）在 4°C 下储存 28 天，其中包含 10%胎牛血清，（iv）在无胎牛血清的 37°C 下储存 28 天，（v）在包含 2%胎牛血清的 37°C 下储存 28 天，或（vi）在包含 10%胎牛血清的 37°C 下储存 28 天。部分样本通过 LIVE/DEAD 染色后进行显微镜分析，以确定软骨细胞活力和密度，包括表面（观察关节表面）和垂直方向（整体和浅层、中层和深层）。其余软骨用于分析硫酸化糖胺聚糖和胶原蛋白。
结果
与 4°C 储存相比，37°C 储存能维持较高的软骨细胞活力。在 37°C 下储存 28 天后，样本在软骨表面的活力约为 80%，在浅层区域约为 65%，在中层区域约为 70%，这明显高于在 4°C 下储存 28 天后的约 45%、约 20%和约 35%，也略高于新鲜对照组的约 100%、约 85%和约 95%。与新鲜对照组相比，37°C 和 4°C 样本的软骨厚度、糖胺聚糖含量和胶原蛋白含量保持不变。
结论
同种异体骨软骨的 37°C 储存可维持软骨细胞的长期活力，特别是在软骨的脆弱表面和浅层区域。
临床意义
同种异体移植物在 37°C 的生理温度下储存可能会延长储存时间，提高移植物的可用性，并改善治疗效果。
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