The effects of cross-linking of collagen-glycosaminoglycan scaffolds on compressive stiffness, chondrocyte-mediated contraction, proliferation and biosynthesis
The effects of cross-linking of collagen-glycosaminoglycan scaffolds on compressive stiffness, chondrocyte-mediated contraction, proliferation and biosynthesis
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DOI：
10.1016/s0142-9612(01)00067-9
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发表时间：
2001-12-01
期刊：
影响因子：
14
通讯作者：
Spector, M
中科院分区：
文献类型：
--
作者：
Lee, CR;Grodzinsky, AJ;Spector, M
使用可植入的三维基质可以改善关节软骨缺损的愈合。研究了四种交联法对胶原-糖胺聚糖(CG)基质压缩硬度的影响以及成年犬关节软骨细胞与基质的相互作用：去水热处理(DHT)、紫外线照射(UV)、戊二醛处理(GTA)和1-乙基-3-(3-二甲氨基丙基)碳二亚胺(EDAC)。在体外四周的时间里，评估软骨细胞介导的收缩、软骨细胞增殖以及蛋白质和糖胺多糖合成的程度和动力学。细胞介导的基质收缩随着交联度的不同而不同：最顺应性的DHT和UV基质收缩最大(基质直径减少60%)和最硬的EDAC。基质收缩最小(基质直径减少30%)。所有的交联方案都允许细胞增殖和基质合成，分别通过DNA含量和放射性标记的硫酸盐和脯氨酸掺入来衡量。在培养的第一周，GTA基质的增殖水平较低，但在四周的培养期间，GTA和EDAC基质提供了最大的细胞增殖。在第2天，H-3-Pro掺入GTA基质中的速率明显较低(p<；0.003)，尽管在随后的时间点，EDAC和GTA基质显示出最高水平的基质合成。关于软骨特异性基质分子的合成，免疫组织化学显示早期DHT和UV基质中有较多的H型胶原。这些发现为进一步研究关节软骨的组织工程学以及软骨细胞收缩与有丝分裂和生物合成过程之间的关系奠定了基础。(C)2001爱思唯尔科学有限公司。保留所有权利。
The healing of articular cartilage defects may be improved by the use of implantable three-dimensional matrices. The present study investigated the effects of four cross-linking methods on the compressive stiffness of collagen-glycosaminoglycan (CG) matrices and the interaction between adult canine articular chondrocytes and the matrix: dehydrothermal treatment (DHT), ultraviolet irradiation (UV), glutaraldehyde treatment (GTA), and 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDAC). The degree and kinetics of chondrocyte-mediated contraction, chondrocyte proliferation, and protein and glycosaminoglycan synthesis were evaluated over a four-week period in vitro. Cell-mediated contraction of the matrices varied with cross-linking: the most compliant DHT and UV matrices contracted the most (60% reduction in matrix diameter) and stiffest EDAC. matrices contracted the least (30% reduction in matrix diameter). All cross-linking protocols permitted cell proliferation and matrix synthesis as measured by DNA content and radiolabeled sulfate and proline incorporation, respectively. During the first week in culture, a lower level of proliferation was seen in the GTA matrices but over the four-week culture period, the GTA and EDAC matrices provided for the greatest cell proliferation. On day 2, there was a significantly lower rate of H-3-proline incorporation in the GTA matrices (p < 0.003) although at later time points, the EDAC and GTA matrices exhibited the highest levels of matrix synthesis. With regard to cartilage-specific matrix molecule synthesis, immunohistochemistry revealed a greater amount of type H collagen in DHT and UV matrices at the early time points. These findings serve as a foundation for future studies of tissue engineering of articular cartilage and the association of chondrocyte contraction and the processes of mitosis and biosynthesis. (C) 2001 Elsevier Science Ltd. All rights reserved.