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(Investigative Ophthalmology and Visual Science. 2002;43:2603-2608.)
© 2002 by The Association for Research in Vision and Ophthalmology, Inc.

Injured Corneal Epithelial Cells Promote Myodifferentiation of Corneal Fibroblasts

Kunihiko Nakamura, Daijiro Kurosaka, Mami Yoshino, Takeshi Oshima and Hiroyo Kurosaka

From the Department of Ophthalmology, Keio University School of Medicine, Tokyo, Japan.

PURPOSE. To determine whether injured corneal epithelial cells stimulate myodifferentiation in corneal fibroblasts and whether transforming growth factor (TGF)-ß is involved.

METHODS. Rabbit corneal fibroblasts were cultured on collagen gel, with or without cocultured corneal epithelial cells or with partially scraped epithelial cells, on a companion plate separated by a permeable membrane. To evaluate fibroblast-induced gel contraction, gel thickness was measured daily relative to the original thickness. Total fibroblasts on the gel were counted. Myofibroblasts were counted by using immunocytochemical identification with anti-{alpha}-smooth muscle actin ({alpha}-SMA). TGF-ß was assayed in the media on days 3 and 6. These procedures also were performed in the presence of anti-TGF-ß antibody.

RESULTS. Gel contraction, {alpha}-SMA-positive cells, and total cell number were significantly greater on gels with injured epithelial cells than on gels without epithelial cells or with uninjured epithelial cells, as was TGF-ß concentration in the media. Anti-TGF-ß antibody eliminated these differences.

CONCLUSIONS. Injured epithelial cells stimulate myodifferentiation in fibroblasts through one or more soluble factors, including TGF-ß.




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