Temporal and spatial expression of matrix metalloproteinases during wound healing of human corneal tissue
Temporal and spatial expression of matrix metalloproteinases during wound healing of human corneal tissue
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DOI：
10.1016/j.exer.2003.08.010
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发表时间：
2003-12-01
影响因子：
3.4
通讯作者：
Saarialho-Kere, U
中科院分区：
文献类型：
--
作者：
Daniels, JT;Geerling, G;Saarialho-Kere, U
我们对角膜修复过程中MMP表达的理解以前依赖于动物模型、分离的人类活检标本和细胞培养研究。本研究的目的是确定培养的人角膜组织创伤后基质金属蛋白酶的时空表达。培养人角膜并切成六片。用角膜刷去除上皮。然后将组织再培养，并将组织片固定至创伤后7天。原位杂交和免疫组化检测基质金属蛋白酶。细胞内层粘连蛋白-5是迁移上皮细胞的标志物，通过免疫组织学定位。在所研究的时间尺度内，来自9个角膜的组织系列实现了上皮细胞对基质的覆盖和受损基底膜的部分修复，这通过高碘酸-希夫反应和苏木精和伊红复染来证明。到第3天，迁移的上皮细胞和基质细胞下方的创伤区表达胶原酶-1（MMP-1）。基质溶解素-1（MMP-3）仅由成纤维样基质细胞表达。基质溶解素-2（MMP-10）在迁移的上皮细胞中被检测到，并且当基质被单层上皮细胞包围时仍然存在。到第7天，组织周围的多层上皮的发展与上皮细胞和基质细胞中MMP表达的停止相一致，除了MMP-9，其仍然存在于上皮基底细胞中。基质金属蛋白酶组织抑制剂-1主要与基质细胞相关，在多层上皮形成后减少。这项研究表明，基质金属蛋白酶的表达在上皮细胞和成纤维细胞样细胞受伤后，人角膜组织在文化中的细胞保持接触，其天然基质。(C)2003年爱思唯尔有限公司保留所有战斗。
Our understanding of MMP expression during corneal repair has previously relied upon animal models, isolated human biopsy specimens and cell culture studies. The aim of this study was to determine the temporal and spatial expression of matrix metalloproteinases following wounding of cultured human corneal tissue. Human corneas were cultured and cut into six pieces. The epithelium was removed with a corneal brush. The tissue was then re-cultured and tissue pieces were fixed up to 7 days post-wounding. Matrix metalloproteinases were detected by in situ hybridisation and immunohistochemistry. Intracellular laminin-5, a marker of migratory epithelial cells, was located immunohistologically. In the time scale studied tissue series from nine corneas achieved coverage of the stroma with epithelial cells and partial repair of damaged basement membrane, demonstrated by the Periodic acid-Schiff reaction and haematoxylin and eosin counter-staining. By day 3, migrating epithelial cells and stromal cells beneath the wounded area expressed collagenase-1 (MMP-1). Stromelysin-1 (MMP-3) was expressed only by fibroblast-like stromal cells. Stromelysin-2 (MMP-10) was detected in migrating epithelial cells and remained when the stroma was surrounded by a monolayer of epithelial cells. By day 7, development of multi-layered epithelium around the tissue coincided with cessation of MMP expression in both epithelial and stromal cells, except for MMP-9, which remained in epithelial basal cells. Tissue inhibitor of matrix metalloproteinase-1 was mainly associated with stromal cells and was reduced upon formation of a multilayered epithelium. This study demonstrates matrix metalloproteinase expression in epithelial and fibroblast-like cells following wounding of human corneal tissue in culture where the cells remain in contact with their natural matrices. (C) 2003 Elsevier Ltd. All fights reserved.