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首页> 外文期刊>Journal of orthopaedic research >Coordinate upregulation of cartilage matrix synthesis in fibrin cultures supplemented with exogenous insulin-like growth factor-I.
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Coordinate upregulation of cartilage matrix synthesis in fibrin cultures supplemented with exogenous insulin-like growth factor-I.

机译：补充外源胰岛素样生长因子-I的纤维蛋白培养物中的软骨基质合成的协调上调。

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The addition of insulin-like growth factor-I to cartilage cultures is known to stimulate the synthesis of proteoglycan and type-II collagen in explant and monolayer studies. The purpose of this study was to determine the effects of long-term supplementation with insulin-like growth factor-I in chondrocytes cultured in fibrin discs as a preliminary investigation to in vivo application of chondrocyte/insulin-like growth factor-I/fibrin grafts to articular-cartilage repair procedures. Chondrocyte-fibrin cultures were maintained for 14 days, with insulin-like growth factor-I added at varying concentrations of 0, 10, 50, or 100 ng/ml medium. Cultures supplemented with 50 or 100 ng of growth factor/ml had increased levels of aggrecan and type-IIB procollagen mRNA, and translation to aggrecan and type-IIB collagen was confirmed by dye-binding assay of total proteoglycan, type-II collagen immunohistochemistry, and determination of collagen content by high-performance liquid chromatography. Maintenance of the chondrocyte phenotype during the 14 days of culture was confirmed by round cell morphology on routine staining, expression of type-II procollagen mRNA on in situ hybridization, evidence of production of pericellular type-II collagen on immunocytochemistry, synthesis of large-molecular-size aggrecan monomer on CL-2B column chromatography, and lack of appreciable message expression for type I or IIA collagen on Northern blot hybridization. Dose-response effects of insulin-like growth factor-I on the expression of chondrocyte matrix constituents were most pronounced at 50 and 100 ng of growth factor per milliliter of medium. These data confirm that (a) culture of chondrocytes for extended periods in three-dimensional cultures of fibrin maintains the chondrocyte phenotype and (b) supplementation with increasing concentrations of insulin-like growth factor-I enhances chondrocyte matrix synthesis and may provide a means to enhance chondrocyte phenotypic stability and function during transplantation grafting procedures.
机译：在外植体和单层研究中，向软骨培养物中添加胰岛素样生长因子-I可以刺激蛋白聚糖和II型胶原蛋白的合成。这项研究的目的是确定长期补充胰岛素样生长因子-I在纤维蛋白盘中培养的软骨细胞中的作用，作为体内应用软骨细胞/胰岛素样生长因子-I /纤维蛋白移植物的初步研究进行关节软骨修复程序。软骨细胞纤维蛋白培养物保持14天，并添加浓度为0、10、50或100 ng / ml的胰岛素样生长因子-I。补充了50或100 ng生长因子/ ml的培养物的聚集蛋白聚糖和IIB型前胶原mRNA水平升高，并且通过总蛋白聚糖的染料结合测定，II型胶原免疫组织化学证实了向聚集蛋白聚糖和IIB型胶原的翻译，高效液相色谱法测定胶原蛋白含量通过常规染色的圆形细胞形态，原位杂交表达II型前胶原mRNA，通过免疫细胞化学产生周细胞II型胶原蛋白的证据，大分子合成的证据，证实了在培养的14天中软骨细胞表型的维持。 CL-2B柱层析上的大号聚集蛋白聚糖单体，Northern blot杂交中I型或IIA型胶原蛋白缺乏明显的信息表达。胰岛素样生长因子-I对软骨细胞基质成分表达的剂量反应效应在每毫升培养基中生长因子分别为50和100 ng时最为明显。这些数据证实了（a）在纤维蛋白的三维培养物中延长软骨细胞的培养可以维持软骨细胞表型，并且（b）补充胰岛素样生长因子-I的浓度增加可以增强软骨细胞基质的合成，并且可以提供一种手段在移植过程中增强软骨细胞表型的稳定性和功能。

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