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  • Title: Corneal epithelial wound healing impaired in keratinocyte-specific HB-EGF-deficient mice in vivo and in vitro.
    Author: Yoshioka R, Shiraishi A, Kobayashi T, Morita S, Hayashi Y, Higashiyama S, Ohashi Y.
    Journal: Invest Ophthalmol Vis Sci; 2010 Nov; 51(11):5630-9. PubMed ID: 20554614.
    Abstract:
    PURPOSE: To study the role played by HB-EGF in corneal epithelial wound healing. METHODS: A 2-mm corneal epithelial wound was created in keratinocyte-specific, HB-EGF-deficient mice--HB(lox/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. RESULTS: 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 HB(lox/+):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. CONCLUSIONS: HB-EGF plays an important role in corneal epithelial cell healing by enhancing cellular attachment and part of cell proliferation.
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