Townley William A, Cambrey Alison D, Khaw Peng T, Grobbelaar Adriaan O
RAFT Institute of Plastic and Reconstructive Surgery, Mount Vernon Hospital, The Leopold Muller Building, Northwood, UK.
J Hand Surg Am. 2008 Nov;33(9):1608-16. doi: 10.1016/j.jhsa.2008.06.013.
Dupuytren's disease is a common fibroproliferative condition of the hand characterized by fibrotic lesions (nodules and cords), leading to disability through progressive digital contracture. Although the etiology of the disease is poorly understood, recent evidence suggests that abnormal matrix metalloproteinase (MMP) activity may play a role in cell-mediated collagen contraction and tissue scarring. The aim of this study was to investigate the efficacy of ilomastat, a broad-spectrum MMP inhibitor, in an in vitro model of Dupuytren fibroblast-mediated contraction.
Nodule-derived and cord-derived fibroblasts were isolated from Dupuytren patients; carpal ligament-derived fibroblasts acted as control. Stress-release fibroblast-populated collagen lattices (FPCLs) were used as a model of contraction. FPCLs were allowed to develop mechanical stress (48 hours) during treatment with ilomastat (0-100 micromol/L), released, and allowed to contract over a 48-hour period. Contraction was estimated by measuring lattice area compared with untreated cells or treatment with a control peptide. MMP-1, MMP-2, and MT1-MMP levels were assessed by zymography, Western blotting, and enzyme-linked immunosorbent assay.
Nodule-derived fibroblasts contracted lattices (69% +/- 2) to a greater extent than did cord-derived (55% +/- 3) or carpal ligament-derived (55% +/- 1) fibroblasts. Exposure to ilomastat led to significant inhibition of lattice contraction by all fibroblasts, although a reduction in lattice contraction by nodule-derived fibroblasts was most prominent (84% +/- 8). In addition, treatment with ilomastat led to a concomitant suppression of MMP-1 and MMP-2 activity, whereas MT1-MMP activity was found to be upregulated.
Our results demonstrate that inhibition of MMP activity results in a reduction in extracellular matrix contraction by Dupuytren fibroblasts and suggest that MMP activity may be a critical target in preventing recurrent contracture caused by this disease.
杜普伊特伦挛缩病是一种常见的手部纤维增生性疾病，其特征为纤维化病变（结节和条索），可通过渐进性手指挛缩导致残疾。尽管该病的病因尚不清楚，但最近的证据表明，异常的基质金属蛋白酶（MMP）活性可能在细胞介导的胶原收缩和组织瘢痕形成中起作用。本研究的目的是在杜普伊特伦成纤维细胞介导的收缩体外模型中研究广谱MMP抑制剂伊洛马司他的疗效。
从杜普伊特伦病患者中分离出结节来源和成条索状来源的成纤维细胞；腕韧带来源的成纤维细胞作为对照。应力释放的成纤维细胞填充胶原晶格（FPCL）用作收缩模型。在用伊洛马司他（0 - 100微摩尔/升）处理期间，使FPCL产生机械应力（48小时），然后释放，并使其在48小时内收缩。通过测量晶格面积并与未处理的细胞或用对照肽处理的细胞进行比较来估计收缩情况。通过酶谱分析、蛋白质印迹和酶联免疫吸附测定来评估MMP - 1、MMP - 2和MT1 - MMP的水平。
结节来源的成纤维细胞使晶格收缩（69%±2）的程度大于条索状来源（55%±3）或腕韧带来源（55%±1）的成纤维细胞。暴露于伊洛马司他导致所有成纤维细胞对晶格收缩的显著抑制，尽管结节来源的成纤维细胞对晶格收缩的降低最为显著（84%±8）。此外，用伊洛马司他处理导致MMP - 1和MMP - 2活性同时受到抑制，而发现MT1 - MMP活性上调。
我们的结果表明，抑制MMP活性可导致杜普伊特伦成纤维细胞引起的细胞外基质收缩减少，并表明MMP活性可能是预防该疾病引起的复发性挛缩的关键靶点。
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