Injured corneal epithelial cells promote myodifferentiation of corneal fibroblasts.
OBJECTIVE
To determine whether injured corneal epithelial cells stimulate myodifferentiation in corneal fibroblasts and whether transforming growth factor (TGF)-beta 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-beta was assayed in the media on days 3 and 6. These procedures also were performed in the presence of anti-TGF-beta 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-beta concentration in the media. Anti-TGF-beta antibody eliminated these differences.


CONCLUSIONS
Injured epithelial cells stimulate myodifferentiation in fibroblasts through one or more soluble factors, including TGF-beta.
Nakamura
Kunihiko
K
Department of Ophthalmology, Keio University School of Medicine, Tokyo, Japan. up4k-nkmr@asahi-net.or.jp
Kurosaka
Daijiro
D
Yoshino
Mami
M
Oshima
Takeshi
T
Kurosaka
Hiroyo
H
D000199:Actins / Q000378:metabolism; D000818:Animals; D002452:Cell Count; D002454:Cell Differentiation / Q000502:physiology*; D002455:Cell Division; D002478:Cells, Cultured; D018920:Coculture Techniques; D003094:Collagen; D003315:Cornea / Q000166:cytology*; D065306:Corneal Injuries; D004797:Enzyme-Linked Immunosorbent Assay; D004847:Epithelial Cells / Q000166:cytology* / Q000378:metabolism; D005347:Fibroblasts / Q000166:cytology* / Q000378:metabolism; D007124:Immunoenzyme Techniques; D011817:Rabbits; D016212:Transforming Growth Factor beta / Q000378:metabolism; D014945:Wound Healing
D000199:Actins; D016212:Transforming Growth Factor beta; D003094:Collagen