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Transforming growth factor‐β3 regulates assembly of a non‐fibrotic matrix in a 3D corneal model
Author(s) -
Karamichos D.,
Hutcheon A. E. K.,
Zieske J. D.
Publication year - 2011
Publication title -
journal of tissue engineering and regenerative medicine
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.835
H-Index - 72
eISSN - 1932-7005
pISSN - 1932-6254
DOI - 10.1002/term.429
Subject(s) - extracellular matrix , chemistry , transforming growth factor , blot , downregulation and upregulation , matrix (chemical analysis) , gene isoform , cornea , transforming growth factor beta , type i collagen , microbiology and biotechnology , biochemistry , biology , ophthalmology , endocrinology , medicine , gene , chromatography
Corneal tissue engineering has attracted the attention of many researchers over the years, in part due to the cornea's avascularity and relatively straightforward structure. However, the highly organized and structured nature of this optically clear tissue has presented a great challenge. We have previously developed a model in which human corneal fibroblasts (HCFs) are stimulated by a stable vitamin C (VitC) derivative to self‐assemble an extracellular matrix (ECM). Addition of TGFβ1 enhanced the assembly of ECM; however, it was accompanied by the upregulation of specific fibrotic markers. In this study, we tested the effects of all three TGFβ isoforms (‐β1, ‐β2 and ‐β3) on ECM production, as well as expression of fibrotic markers. HCFs were grown in four media conditions for 4 weeks: control, VitC only; T1, VitC + TGFβ1; T2, VitC + TGFβ2; and T3, VitC + TGFβ3. The cultures were analysed with western blots, TEM and indirect immunofluorescence (IF). Compared to controls, all TGFβ isoforms stimulated matrix production by about three‐fold. IF showed the presence of type III collagen and smooth muscle actin (SMA) in T1 and T2; however, T3 showed little to no expression. In western blots, T3 stimulated a lower type III:type I collagen ratio when compared to the other conditions. In addition, TEM indicated that T3 stimulated a higher level of matrix alignment and organization. HCFs stimulated by VitC and TGFβ3 appear to generate a matrix that mimics the normal adult or developing human cornea, whereas TGF‐β1 and ‐β2 drive the constructs towards a more fibrotic path. Copyright © 2011 John Wiley & Sons, Ltd.