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Ryuji Yoshioka, Atsushi Shiraishi, Takeshi Kobayashi, Shin-ichi Morita, Yasuhito Hayashi, Shigeki Higashiyama, Yuichi Ohashi; Corneal Epithelial Wound Healing Impaired in Keratinocyte-Specific HB-EGF–Deficient Mice In Vivo and In Vitro. Invest. Ophthalmol. Vis. Sci. 2010;51(11):5630-5639. doi: 10.1167/iovs.10-5158.
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© 2017 Association for Research in Vision and Ophthalmology.
To study the role played by HB-EGF in corneal epithelial wound healing.
A 2-mm corneal epithelial wound was created in keratinocyte-specific, HB-EGF–deficient mice—HBlox/lox:K-5Cre (HB−/−)—and the speed of wound healing was compared with that in wild-type (WT) mice. Cultured confluent mouse corneal epithelial cells (MCECs) from WT and HB−/− mice were scraped, and the bare area was measured. The proliferation of MCECs was determined by BrdU incorporation. The degree of attachment of WT and HB−/− MCECs was also determined. The mRNA expression of EGF family and cell adhesion molecules was determined by real-time PCR.
Corneal epithelial wound healing was significantly delayed in HB−/− mice, and the expression of HB-EGF was detected at the leading edge of the wound in HBlox/+:K5-Cre (HB+/−) mice by the presence of lacZ staining. The wound closure was significantly impaired in HB−/− MCECs and was improved by adding HB-EGF. The number of BrdU-positive MCECs of WT and HB−/− mice was not significantly different, and both increased to different degrees when HB-EGF was added. The adhesion of isolated HB−/− MCECs was lower than that of WT MCECs, but the degree of adhesion was restored by adding HB-EGF. The expression of epiregulin was upregulated in HB−/− MCECs, and α6- and β1-integrin were upregulated by adding HB-EGF.
HB-EGF plays an important role in corneal epithelial cell healing by enhancing cellular attachment and part of cell proliferation.
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