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含表没食子儿茶素没食子酸酯的冷藏溶液保存的同种异体骨软骨的生物学和生物力学评估。

Biological and biomechanical evaluations of osteochondral allografts preserved in cold storage solution containing epigallocatechin gallate.

机构信息

Department of Medical Simulation Engineering, Research Center for Nano Medical Engineering, Institute for Frontier Medical Sciences, Kyoto University, Kyoto, Japan.

出版信息

Cell Transplant. 2010;19(6):681-9. doi: 10.3727/096368910X508771. Epub 2010 Jun 3.

Abstract

The beneficial effects of (-)-epigallocatechin-3-O-gallate (EGCG) on the nonfrozen preservation of mammalian cells and tissues are generally not well understood. A storage solution containing EGCG was employed to test the hypothesis that EGCG is capable of extending the storage duration for the cold preservation of articular cartilages. Human articular cartilages were preserved in a storage solution composed of serum-free RPMI-1640 medium with 1% antibiotic-antimycotic solution and 1 mM EGCG at 4°C for 1, 2, and 4 weeks. The chondrocyte viability (CCK-8 assay), biochemical and immunohistochemical composition [glycosaminoglycans (GAG) and (type II) collagen], and biomechanical property (compressive elastic modulus) were assessed. The chondrocyte viability of the cartilages preserved with EGCG was significantly well maintained for at least 2 weeks with high content of GAG and total collagen. These beneficial effects of EGCG were confirmed by the immunohistochemical observations of well-preserved cartilaginous structures and delayed denaturation of the extracellular matrix in preserved cartilages. There was no significant difference in the compressive elastic modulus (MPa) between the cartilages preserved with and without EGCG. These results suggest that EGCG may play an effective role in preserving osteochondral allografts, which can be exploited in devising strategies for the long-term preservation of other tissues under cold storage conditions.

摘要

(-)-表没食子儿茶素没食子酸酯(EGCG)对哺乳动物细胞和组织的非冷冻保存的有益作用通常不太了解。采用含有 EGCG 的储存溶液来检验假设,即 EGCG 能够延长关节软骨的冷藏保存时间。将人关节软骨保存在含有无血清 RPMI-1640 培养基、1%抗生素-抗真菌溶液和 1mM EGCG 的储存溶液中,在 4°C 下分别保存 1、2 和 4 周。通过 CCK-8 测定法评估软骨细胞活力、生物化学和免疫组织化学组成[糖胺聚糖(GAG)和(II 型)胶原蛋白]以及生物力学特性(压缩弹性模量)。用 EGCG 保存的软骨的软骨细胞活力至少在 2 周内保持良好,且 GAG 和总胶原蛋白含量高。保存的软骨中软骨结构保存良好以及细胞外基质变性延迟的免疫组织化学观察结果证实了 EGCG 的这些有益作用。用 EGCG 和不用 EGCG 保存的软骨之间的压缩弹性模量(MPa)没有显著差异。这些结果表明,EGCG 可能在保存同种异体骨软骨移植物中发挥有效作用,这可用于设计在冷藏条件下长期保存其他组织的策略。

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