van der Kraan P M, Vitters E L, Meijers T H, Poole A R, van den Berg W B
Department of Rheumatology, University Hospital Nijmegen, The Netherlands.
Osteoarthritis Cartilage. 1998 Nov;6(6):417-26. doi: 10.1053/joca.1998.0145.
Articular cartilage has only limited capacities for repair and it is not known what is the exact mechanism of matrix restoration. It was investigated whether the reparative process in murine articular cartilage after moderate proteoglycan depletion is accompanied by a change in the chondrocyte phenotype either to hypertrophy or to a less differentiated phenotype as assayed by the expression of specific collagen subtypes.
Moderate proteoglycan depletion was induced by injection of papain whereafter the expression of collagen type I mRNA, collagen IIA and IIB mRNA and type X collagen mRNA in patellar cartilage, as markers for chondrocyte phenotype, was investigated by RT-PCR during normal cartilage physiology and matrix restoration. In addition, in-situ expression of collagen subtypes was assayed by immunolocalisation.
In normal articular cartilage collagen I, collagen IIB and collagen type X transcripts were easily detected. Surprisingly, collagen type I sense as well as antisense mRNA was detected and in addition to IIB transcripts collagen IIA transcripts were detected in a number of samples. During cartilage matrix restoration no change in the expression of collagen I, collagen IIA or IIB or collagen type X mRNA transcripts could be detected. Immunolocalization demonstrated the presence of type I (pericellular) and type II collagen in the extracellular matrix. The pericellular matrix of hypertrophic chondrocytes showed collagen type X staining in the calcified cartilage in normal and papain-injected knee joints. Increased staining for collagen type X was found in the upper cartilage layer in the interterritorial matrix from day 7 after papain injection.
The absence of changes in collagen mRNA expression indicates that alteration of chondrocyte phenotype does not occur during the successful repair process after moderate proteoglycan depletion. Collagen type X appears to be deposited in the upper cartilage layer during this process.
关节软骨的修复能力有限,目前尚不清楚基质修复的确切机制。本研究旨在探讨在适度蛋白聚糖耗竭后,小鼠关节软骨的修复过程中,软骨细胞表型是否会转变为肥大或分化程度较低的表型,这可通过特定胶原亚型的表达来检测。
通过注射木瓜蛋白酶诱导适度的蛋白聚糖耗竭,随后在正常软骨生理和基质修复过程中,利用逆转录聚合酶链反应(RT-PCR)研究髌软骨中I型胶原mRNA、IIA和IIB型胶原mRNA以及X型胶原mRNA的表达,以此作为软骨细胞表型的标志物。此外,通过免疫定位检测胶原亚型的原位表达。
在正常关节软骨中,I型胶原、IIB型胶原和X型胶原转录本易于检测到。令人惊讶的是,检测到了I型胶原的正义和反义mRNA,并且在一些样本中除了IIB转录本外还检测到了IIA型胶原转录本。在软骨基质修复过程中,未检测到I型胶原、IIA型胶原、IIB型胶原或X型胶原mRNA转录本表达的变化。免疫定位显示细胞外基质中存在I型(细胞周围)和II型胶原。在正常和注射木瓜蛋白酶的膝关节中,肥大软骨细胞的细胞周围基质在钙化软骨中显示X型胶原染色。在注射木瓜蛋白酶后第7天,在区域间基质的上层软骨层中发现X型胶原染色增加。
胶原mRNA表达未发生变化表明,在适度蛋白聚糖耗竭后的成功修复过程中,软骨细胞表型未发生改变。在此过程中,X型胶原似乎沉积在上层软骨层中。