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Corneal epithelial wound healing impaired in keratinocyte-specific HB-EGF-deficient mice in vivo and in vitro.

AbstractPURPOSE:
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.
AuthorsRyuji Yoshioka, Atsushi Shiraishi, Takeshi Kobayashi, Shin-Ichi Morita, Yasuhito Hayashi, Shigeki Higashiyama, Yuichi Ohashi
JournalInvestigative ophthalmology & visual science (Invest Ophthalmol Vis Sci) Vol. 51 Issue 11 Pg. 5630-9 (Nov 2010) ISSN: 1552-5783 [Electronic] United States
PMID20554614 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
Chemical References
  • Cell Adhesion Molecules
  • Hbegf protein, mouse
  • Heparin-binding EGF-like Growth Factor
  • Intercellular Signaling Peptides and Proteins
  • RNA, Messenger
  • Epidermal Growth Factor
Topics
  • Animals
  • Cell Adhesion Molecules (genetics)
  • Cell Proliferation
  • Cells, Cultured
  • Epidermal Growth Factor (genetics)
  • Epithelium, Corneal (cytology)
  • Heparin-binding EGF-like Growth Factor
  • Intercellular Signaling Peptides and Proteins (physiology)
  • Keratinocytes (metabolism)
  • Mice
  • Mice, Inbred C57BL
  • Mice, Knockout
  • Mice, Transgenic
  • RNA, Messenger (genetics)
  • Reverse Transcriptase Polymerase Chain Reaction
  • Wound Healing (physiology)

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