Diefenderfer D L, Brighton C T
McKay Laboratories for Orthopaedic Surgery Research, Department of Orthopaedic Surgery, School of Medicine, University of Pennsylvania, 36th and Hamilton Walk, Philadelphia, Pennsylvania 19104-6081, USA.
Biochem Biophys Res Commun. 2000 Mar 5;269(1):172-8. doi: 10.1006/bbrc.2000.2148.
Multipotential mesenchymal stem cells capable of chondro-osseous induction contribute to the endochondral callus of healing fractured bone. Microvascular pericytes serving the role of multipotential mesenchymal stem cells are considered osteoprogenitors because they express type I collagen, alkaline phosphatase enzyme activity, osteocalcin immunoreactivity, and bone sialoprotein mRNA. Previous electron microscopic studies indicate that this cell type has a contribution to the fracture callus. Limited data suggest that pericytes may also assume a chondrogenic phenotype. We undertook in vitro studies to understand how the chondro-osseous phenotype of the pericyte might be regulated. Using Northern analysis and semiquantitative reverse transcriptase-polymerase chain reaction (RT-PCR), we found that cultured pericytes produce aggrecan and type II collagen mRNA indicating their chondrogenic potential. Aggrecan message is elevated by BMP-2 as analyzed by both Northern hybridization and RT-PCR. This finding suggests a regulatory role for this morphogen on this phenotype in pericytes. RT-PCR amplified versican product was also associated with pericyte cultures but was not affected by BMP-2. Our data strongly support a chondrogenic role for the pericyte and that the phenotype is regulated at least in part by BMP.
具有软骨-骨诱导能力的多能间充质干细胞有助于愈合骨折的软骨内骨痂形成。充当多能间充质干细胞角色的微血管周细胞被认为是骨祖细胞,因为它们表达I型胶原、碱性磷酸酶活性、骨钙素免疫反应性和骨唾液蛋白mRNA。先前的电子显微镜研究表明,这种细胞类型对骨折骨痂有贡献。有限的数据表明,周细胞也可能呈现软骨生成表型。我们进行了体外研究,以了解周细胞的软骨-骨表型可能是如何被调控的。通过Northern印迹分析和半定量逆转录-聚合酶链反应(RT-PCR),我们发现培养的周细胞产生聚集蛋白聚糖和II型胶原mRNA,表明它们具有软骨生成潜力。通过Northern杂交和RT-PCR分析发现,BMP-2可使聚集蛋白聚糖信息增加。这一发现提示这种形态发生素对周细胞的这种表型具有调控作用。RT-PCR扩增的多功能蛋白聚糖产物也与周细胞培养物相关,但不受BMP-2影响。我们的数据有力地支持了周细胞的软骨生成作用,并且该表型至少部分受BMP调控。