Plastic and Reconstructive Surgery Research, Manchester Interdisciplinary Biocentre, University of Manchester, Manchester, United Kingdom.
Stem Cells Dev. 2012 Mar 1;21(4):609-22. doi: 10.1089/scd.2011.0140. Epub 2011 Jul 19.
Dupuytren's disease (DD) is a fibroproliferative disorder characterized by aberrant proliferation of myofibroblasts, the source of which remains unknown. Recent studies indicate that circulating and tissue-resident mesenchymal stem cells (MSCs) can differentiate into myofibroblasts. Therefore, the aim of this study was to profile MSCs from phenotypically distinct DD sites including cord, nodule, skin overlying nodule (SON), and perinodular fat (PNF) compared with unaffected internal controls, that is, distant palmar fat (DPF) and transverse palmar fascia (Skoog's fibers) as well as external control carpal tunnel (CT) tissue including skin, fat, and fascia. Freshly isolated primary fibroblasts as well as cells grown up to passage 5 (P5) from DD (n=27) and CT (n=14) samples were analyzed for the presence of established MSC markers CD73, CD90, and CD105 and absence of hematopoietic marker CD34 using fluorescence-activated cell sorting, in-cell quantitative western blotting, immunohistochemistry, and immunocytochemistry. Freshly isolated cells from SON, PNF, and cord biopsies had a higher number of CD34(-)73(+)90(+)105(+) cells compared with Skoog's fibers and CT controls. P3 cells obtained from all DD biopsies compared with CT samples differentiated into osteocytes, adipocytes, and chondrocytes. P3 cord and nodule cells expressed intense α-smooth muscle actin staining compared with skin and fat cells. Stem cell markers including stem cell factor, MSC-homing marker CXCR4, and Wnt/β-catenin downregulator Dkk-1 were all upregulated in SON and PNF compared with CT skin and CT fat, respectively, as shown by real-time quantitative polymerase chain reaction. However, osteogenic marker OSF-1 had a significantly higher expression in the PNF (P=0.002) and cord (P=0.01) compared with the nodule. In conclusion, we have shown the presence of MSCs in specific DD tissue phenotypes compared with internal and external control tissue. These findings provide preliminary support for a potential alternative source of disease myofibroblasts originating from sites such as SON and PNF as opposed to palmar fascia alone.
掌腱膜挛缩症 (Dupuytren's disease, DD) 是一种纤维增生性疾病，其特征是肌成纤维细胞异常增殖，但其来源尚不清楚。最近的研究表明，循环和组织驻留的间充质干细胞 (mesenchymal stem cells, MSCs) 可以分化为肌成纤维细胞。因此，本研究旨在对表型不同的 DD 部位（包括 cord、nodule、son、PNF）的 MSCs 进行分析，这些部位与未受影响的内部对照（即远端掌脂垫 DPF、掌腱膜横纤维 Skoog's fibers）以及外部对照腕管 CT 组织（包括皮肤、脂肪和筋膜）进行比较。使用流式细胞术、细胞内定量 Western blot、免疫组化和免疫细胞化学分析从 DD（n=27）和 CT（n=14）样本中分离的新鲜原代成纤维细胞以及培养至第 5 代（P5）的细胞中，检测已建立的 MSC 标志物 CD73、CD90 和 CD105 的表达情况，以及造血标志物 CD34 的缺失情况。与 Skoog's fibers 和 CT 对照相比，son、PNF 和 cord 活检的新鲜分离细胞具有更高数量的 CD34(-)73(+)90(+)105(+)细胞。与 CT 样本相比，从所有 DD 活检中获得的 P3 细胞可分化为成骨细胞、脂肪细胞和成软骨细胞。与皮肤和脂肪细胞相比，P3 cord 和 nodule 细胞表达强烈的α-平滑肌肌动蛋白染色。实时定量聚合酶链反应显示，与 CT 皮肤和 CT 脂肪相比，SON 和 PNF 中干细胞因子、MSC 归巢标志物 CXCR4 和 Wnt/β-catenin 下调因子 Dkk-1 等干细胞标志物均上调。然而，在 PNF（P=0.002）和 cord（P=0.01）中，成骨标志物 OSF-1 的表达明显高于 nodule。总之，与内部和外部对照组织相比，我们在特定的 DD 组织表型中发现了 MSCs 的存在。这些发现初步支持了源自 son 和 PNF 等部位的疾病肌成纤维细胞的潜在替代来源，而不仅仅是掌腱膜。
