Quaglino D, Bergamini G, Croce A, Boraldi F, Barbieri D, Caroli A, Marcuzzi A, Tiozzo R, Ronchetti I P
Department of Biomedical Sciences, University of Modena, Italy.
J Cell Physiol. 1997 Dec;173(3):415-22. doi: 10.1002/(SICI)1097-4652(199712)173:3<415::AID-JCP14>3.0.CO;2-A.
The present investigation has been performed to better characterize, in vitro, normal aponeurotic cells in comparison with dermal fibroblasts and with cells derived from Dupuytren's affected aponeuroses. Cells were cultured in monolayer and/or into three-dimensional collagen gels. Cell structure, adhesion, and spreading capability on different substrates, as well as integrin expression were investigated by light and electron microscopy and by flow cytometry. Cell-matrix interactions were also analyzed by gel retraction experiments in the presence, or absence, of RGD peptides and anti-integrin antibodies. Normal aponeurotic cells, compared with dermal fibroblasts, exhibited in vitro peculiar structural features, which were substantially maintained in Dupuytren's aponeurotic cells, irrespective of the substrate they were grown on. By contrast, the aponeurotic cell behavior was different in normal and diseased cells, these latter approaching that of dermal fibroblasts. Normal aponeurotic cells, in fact, were characterized by low efficiency in retracting the collagen gel, low alpha 2, alpha 1, and alpha 5 integrin subunit expression and low adhesion properties onto collagen and fibronectin, whereas cells isolated from the aponeuroses of Dupuytren's patients exhibited higher capability of retracting the collagen gel, increased adhesion properties toward collagen and fibronectin, and higher levels of integrin expression. No differences were observed between dermal fibroblasts from Dupuytren's patients or from normal subjects. These in vitro results are consistent with those previously obtained in situ, suggesting that palmar aponeurotic cells have a peculiar phenotype and that changes in cell-matrix interactions occur in Dupuytren's contracture. Moreover, by comparing data obtained from the retracted fibrotic cords and the still clinically unaffected aponeuroses of the same patients, it may be noted that Dupuytren's disease is not only confined to the clinically involved branches, but includes the whole aponeurosis of the affected hand.
本研究旨在更好地在体外对正常腱膜细胞进行表征,并与真皮成纤维细胞以及来自掌腱膜挛缩症患者受累腱膜的细胞进行比较。细胞在单层培养和/或三维胶原凝胶中培养。通过光学显微镜、电子显微镜和流式细胞术研究细胞结构、在不同底物上的黏附及铺展能力,以及整合素表达。还通过在存在或不存在RGD肽和抗整合素抗体的情况下进行凝胶收缩实验来分析细胞-基质相互作用。与真皮成纤维细胞相比,正常腱膜细胞在体外表现出独特的结构特征,无论其生长在何种底物上,这些特征在掌腱膜挛缩症患者的腱膜细胞中基本保持。相比之下,正常和患病细胞中的腱膜细胞行为有所不同,患病细胞的行为更接近真皮成纤维细胞。事实上,正常腱膜细胞的特征在于胶原凝胶收缩效率低、α2、α1和α5整合素亚基表达低以及对胶原和纤连蛋白的黏附特性低,而从掌腱膜挛缩症患者的腱膜中分离出的细胞表现出更高的胶原凝胶收缩能力、对胶原和纤连蛋白的黏附特性增加以及更高水平的整合素表达。在掌腱膜挛缩症患者或正常受试者的真皮成纤维细胞之间未观察到差异。这些体外结果与先前原位获得的结果一致,表明掌腱膜细胞具有独特的表型,并且在掌腱膜挛缩症中细胞-基质相互作用发生了变化。此外,通过比较从同一患者的纤维化挛缩索条和临床上仍未受影响的腱膜获得的数据,可以注意到掌腱膜挛缩症不仅局限于临床上受累的分支,还包括患手的整个腱膜。