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在I型或II型胶原蛋白支架中培养的关节软骨细胞的比较表型分析。

Comparative phenotypic analysis of articular chondrocytes cultured within type I or type II collagen scaffolds.

作者信息

Freyria Anne-Marie, Ronzière Marie-Claire, Cortial Delphine, Galois Laurent, Hartmann Daniel, Herbage Daniel, Mallein-Gerin Frédéric

机构信息

Institut de Biologie et Chimie des Protéines, CNRS UMR 5086, Université de Lyon, IFR 128, Lyon, France.

出版信息

Tissue Eng Part A. 2009 Jun;15(6):1233-45. doi: 10.1089/ten.tea.2008.0114.

Abstract

Among the existing repair strategies for cartilage injury, tissue engineering approach using biomaterials and chondrocytes offers hope for treatments. In this context, collagen-based biomaterials are good candidates as scaffolds for chondrocytes in cell transplantation procedures. These scaffolds are provided under different forms (gel or crosslinked sponge) made with either type I collagen or type I or type II atelocollagen molecules. The present study was undertaken to investigate how bovine articular chondrocytes sense and respond to differences in the structure and organization of these collagen scaffolds, over a 12-day culture period. When chondrocytes were seeded in the collagen scaffolds maintained in free-floating conditions, cells contracted gels to 40-60% and sponges to 15% of their original diameter. Real-time polymerase chain reaction analysis indicated that the chondrocyte phenotype, assessed notably by the ratio of COL2A1/COL1A2 mRNA and alpha10/alpha11 integrin subunit mRNA, was comparatively better sustained in type I collagen sponges when seeded at high cell density, also in type I atelocollagen gels. Besides, proteoglycan accumulation in the different scaffolds, as assessed by measuring the sulfated glycosaminoglycan content, was found be highest in type I collagen sponges seeded at high cell density. In addition, gene expression of matrix metalloproteinase-13 increased dramatically (up to 90-fold) in chondrocytes cultured in the different gels, whereas it remained stable in the sponges. Our data taken together reveal that type I collagen sponges seeded at high cell density represent a suitable material for tissue engineering of cartilage.

摘要

在现有的软骨损伤修复策略中,利用生物材料和软骨细胞的组织工程方法为治疗带来了希望。在这种情况下,基于胶原蛋白的生物材料是细胞移植过程中软骨细胞支架的良好候选材料。这些支架有不同的形式(凝胶或交联海绵),由I型胶原蛋白或I型或II型去端胶原蛋白分子制成。本研究旨在调查在12天的培养期内,牛关节软骨细胞如何感知并响应这些胶原蛋白支架在结构和组织上的差异。当软骨细胞接种到保持自由漂浮状态的胶原蛋白支架中时,细胞将凝胶收缩至其原始直径的40%-60%,将海绵收缩至15%。实时聚合酶链反应分析表明,当以高细胞密度接种时,软骨细胞表型(特别是通过COL2A1/COL1A2 mRNA和α10/α11整合素亚基mRNA的比率评估)在I型胶原蛋白海绵中以及I型去端胶原蛋白凝胶中得到了相对更好的维持。此外,通过测量硫酸化糖胺聚糖含量评估,不同支架中的蛋白聚糖积累在高细胞密度接种的I型胶原蛋白海绵中最高。此外,在不同凝胶中培养的软骨细胞中,基质金属蛋白酶-13的基因表达急剧增加(高达90倍),而在海绵中则保持稳定。综合我们的数据表明,高细胞密度接种的I型胶原蛋白海绵是软骨组织工程的合适材料。

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