Chondrogenic differentiation of growth factor-stimulated precursor cells in cartilage repair tissue is associated with increased HIF-1α activity
Chondrogenic differentiation of growth factor-stimulated precursor cells in cartilage repair tissue is associated with increased HIF-1α activity
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DOI：
10.1016/j.joca.2008.04.006
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发表时间：
2008-12-01
影响因子：
7
通讯作者：
Schneider, H.
中科院分区：
文献类型：
--
作者：
Gelse, K.;Muehle, C.;Schneider, H.
目的：研究生长因子刺激的骨膜细胞在低氧诱导因子1 α活性方面的软骨形成潜力。方法：应用腺相关病毒（AAV）和腺病毒（Ad）载体将胰岛素样生长因子1（IGF-1）或骨形态发生蛋白2（BMP-2）基因转染激活的支架结合自体骨膜细胞应用于小型猪膝关节软骨损伤。移植后6周，研究修复组织的I型和II型胶原含量以及HIF-1 α。表情通过使用特异性抑制剂在体外评估磷脂酰肌醇3-激酶（PI 3 K）/雷帕霉素哺乳动物靶蛋白（mTOR）和丝裂原活化蛋白激酶（MEK）/细胞外信号调节激酶（ERK）信号传导对BMP-2/IGF-1诱导的HIF-1 α表达的功能作用。未受刺激的骨膜细胞在修复组织的浅区形成纤维细胞外基质，在深区形成纤维软骨基质。这种区域差异反映在表面区域缺乏HIF-1 α染色，但中度HIF-1 α染色。在深层表达。相比之下，Ad/AAVBMP-2刺激的骨膜细胞，以及在较小程度上Ad/AAVIGF-1感染的细胞，采用了具有强细胞内HIF-lj的软骨细胞样表型。染色遍及修复组织的所有区域并形成透明样基质。在体外，BMP-2和IGF-1的补充增加了骨膜细胞中HIF-1 α的蛋白水平，这是基于转录后机制，而不是从头mRNA合成，主要涉及IVIEK/ERK途径。对相对少量动物的初步实验研究表明，前体细胞的软骨形成在软骨修复组织的较深缺氧区中得到促进，并受到以下刺激：增强HIF-1 α活性的生长因子。(C)2008年国际骨关节炎研究学会。由爱思唯尔有限公司出版。保留所有权利。
Objective: To investigate the chondrogenic potential of growth factor-stimulated periosteal cells with respect to the activity of Hypoxia-inducible Factor 1 alpha. (HIF-1 alpha).Methods: Scaffold-bound autologous periosteal cells, which had been activated by Insulin-like Growth Factor 1 (IGF-1) or Bone Morphogenetic Protein 2 (BMP-2) gene transfer using both adeno-associated virus (AAV) and adenoviral (Ad) vectors, were applied to chondral lesions in the knee joints of miniature pigs. Six weeks after transplantation, the repair tissues were investigated for collagen type I and type II content as well as for HIF-1 alpha. expression. The functional role of phosphatidylinositol 3-kinase (PI3K)/mammalian target of rapamycin (mTOR) and mitogen-activated protein kinase (MEK)/extracellular signal-regulated kinase (ERK) signaling on BMP-2/IGF-1-induced HIF-1 alpha expression was assessed in vitro by employing specific inhibitors.Results: Unstimulated periosteal cells formed a fibrous extracellular matrix in the superficial zone and a fibrocartilaginous matrix in deep zones of the repair tissue. This zonal difference was reflected by the absence of HIF-1 alpha staining in superficial areas, but moderate HIF-1 alpha. expression in deep zones. In contrast, Ad/AAVBMP-2-stimulated periosteal cells, and to a lesser degree Ad/AAVIGF-1-infected cells, adopted a chondrocyte-like phenotype with strong intracellular HIF-lj. staining throughout all zones of the repair tissue and formed a hyaline-like matrix. In vitro, BMP-2 and IGF-1 supplementation increased HIF-1 alpha, protein levels in periosteal cells, which was based on posttranscriptional mechanisms rather than de novo mRNA synthesis, involving predominantly the IVIEK/ERK pathway.Conclusion: This pilot experimental study on a relatively small number of animals indicated that chondrogenesis by precursor cells is facilitated in deeper hypoxic zones of cartilage repair tissue and is stimulated by growth factors which enhance HIF-1 alpha activity. (C) 2008 Osteoarthritis Research Society International. Published by Elsevier Ltd. All rights reserved.