期刊
INVESTIGATIVE OPHTHALMOLOGY & VISUAL SCIENCE
卷 53, 期 3, 页码 1077-1086出版社
ASSOC RESEARCH VISION OPHTHALMOLOGY INC
DOI: 10.1167/iovs.11-8609
关键词
-
资金
- Research to Prevent Blindness, Inc., New York, NY
- [NIH R01 EY 013322]
- [NIH P30 EY020799]
PURPOSE. To assess how wound healing cytokines and the extracellular matrix (ECM) environment regulate the keratocyte mechanical phenotype. METHODS. Rabbit corneal keratocytes were plated within standard bovine or rat tail type I collagen matrices (2.5 mg/mL), compressed collagen matrices (approximately 100 mg/mL), or on collagen-coated dishes and cultured for up to 7 days in serum-free media, platelet derived growth factor BB (PDGF BB), insulin-like growth factor (IGF), TGF beta 1, TGF beta 2, or FGF2. F-actin, alpha-smooth muscle actin (alpha-SMA) and collagen fibrils were imaged using confocal microscopy. Cell morphology, local matrix reorganization, and global matrix contraction were quantified digitally. RESULTS. IGF and PDGF BB stimulated elongation of keratocytes and extension of dendritic processes within 3-D matrices, without inducing stress fiber formation or collagen reorganization. In contrast, treatment with TGF beta 1 and TGF beta 2 increased keratocyte contractility, as indicated by stress fiber formation and matrix compaction and alignment. This transformation was enhanced at higher cell densities within standard 3-D matrices, in which alpha-SMA was incorporated into stress fibers. In contrast, alpha-SMA was expressed within compressed 3-D matrices even at low cell density. FGF2 did not produce significant cytoskeletal or matrix reorganization in standard 3-D matrices; however, stress fibers were consistently expressed within compressed collagen matrices and on rigid two-dimensional substrates. Inhibiting Rho kinase blocked both TGF beta- and FGF2-induced stress fiber formation. CONCLUSIONS. Keratocytes cultured in IGF or PDGF BB maintain a quiescent mechanical phenotype over a range of matrix environments. In contrast, the mechanical phenotypes induced by FGF and TGF beta vary in response to the structural and/or mechanical properties of the ECM. (Invest Ophthalmol Vis Sci. 2012;53:1077-1086) DOI:10.1167/iovs.11-8609
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