Previous studies suggest the existence of two separate and distinct mechanisms of endothelial wound healing (i.e., cell migration and cell spreading), which may be controlled by unique, injury-dependent, wound- related factors. The purpose of our study was to evaluate potential biologic mediators regulating healing of the growth arrested cat endothelium by using an ex vivo, organ-culture model. Three buttons were punched from each cornea of 11 cats with a 6-mm trephine. A 1- to 2-mm diameter endothelial scrape injury (SI) was made, and buttons were cultured in (a) serum-free media (SFM), (b) serum plus media (20% fetal calf serum), (c) SFM plus basic fibroblast growth factor (bFGF), (d) SFM plus bFGF and heparin, (e) SFM plus transforming growth factor-β1 (TGF(β1)), or (f) SFM plus TGF(β1) and anti-TGF(β1). At various times from 8-48 h after injury, buttons were stained with phalloidin and anti-ZO-1, and imaged by using laser scanning confocal microscopy. Evaluation of SI in cat corneal buttons under serum- free conditions showed maintenance of normal endothelial differentiation, indicating that the organ-culture SI model mimics in vivo SI. Addition of TGF(β1) produced a dramatic reorganization of apical F-actin and development of stress fibers, as well as the loss of normal cell border- associated ZO-1 distribution. The effects of TGF(β1) were blocked by the neutralizing antibodies to TGF(β1). Addition of serum or bFGF produced much less pronounced changes in F-actin and ZO-1 distribution. These results suggest that TGF(β1) may play a critical role in modulating the wound- healing response of the corneal endothelium.
|Original language||English (US)|
|Number of pages||8|
|State||Published - Aug 27 1996|
- Basic fibroblast growth factor
- Endothelial wound healing
- Transforming growth factor-β1
ASJC Scopus subject areas