May 2004
Volume 45, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2004
Thymosin beta 4 Expression In Corneal Wound Healing
Author Affiliations & Notes
  • G. Sosne
    Kresge Eye Institute/Ophthalmology and Anatomy/Cell Biology,
    Wayne State University, Detroit, MI
  • A.L. Goldstein
    Biochemistry and Molecular Biology, George Washington University School of Medicine, Washington, DC
  • M. Badamchian
    Biochemistry and Molecular Biology, George Washington University School of Medicine, Washington, DC
  • R.P. Barrett
    Anatomy/Cell Biology,
    Wayne State University, Detroit, MI
  • L.D. Hazlett
    Anatomy/Cell Biology,
    Wayne State University, Detroit, MI
  • Footnotes
    Commercial Relationships  G. Sosne, None; A.L. Goldstein, RegeneRx Pharmaceuticals I, C; M. Badamchian, None; R.P. Barrett, None; L.D. Hazlett, None.
  • Footnotes
    Support  NEI KO8 EY 13412
Investigative Ophthalmology & Visual Science May 2004, Vol.45, 1425. doi:
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      G. Sosne, A.L. Goldstein, M. Badamchian, R.P. Barrett, L.D. Hazlett; Thymosin beta 4 Expression In Corneal Wound Healing . Invest. Ophthalmol. Vis. Sci. 2004;45(13):1425.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Abstract: : Purpose: The purpose of these studies was to localize and to quantify the protein levels of thymosin beta 4 (Tß4) in the corneas of young vs old mice corneas after wounding. Methods:Corneas from young (6–8 weeks of age) and old (12–13 months of age) ICR mice were debrided with a razor blade to create a 2 mm size epithelial defect. A time course study was performed to examine the rates of corneal healing in the young vs old mice. Animals (n=10/group/time point) were sacrificed at various time points (0, 6, 24 and 48 hours) after wounding and the corneal buttons were excised and assayed by ELISA (ALPCO, New Hampshire) for Tß4 quantification. In addition, whole eyes (n=3/group/time point) from 0, 24, and 48 hours time points were harvested for corneal immunohistochemical (IHC) analysis of Tß4. Results: Young corneas re–epithelialized by 20 hours after wounding, 4 hours earlier than those in the aged mice. Normal, unwounded corneas from the young and aged animals demonstrated similar Tß4 protein levels (2.5 µg/ml cornea). After wounding, the old and young animals both exhibited increases in Tß4 protein levels compared to their respective normal controls but the aged corneal Tß4 expression levels were markedly blunted compared to those in the young. Immediately after wounding, young corneal Tß4 levels were almost 2.5 times more than the aged. These levels peaked at 24 hours when young corneas measured over three–fold more Tß4 than the aged (17.5 µg/ml and 5 µg/ml cornea, respectively) and they decreased at 48 hours (12.5 µg/ml and 4 µg/ml cornea, respectively). The IHC staining in the young corneas was more intense and regular in the epithelial nuclei and at the leading wound edges compared to the aged. The aged corneas localized Tß4 more to the epithelial and endothelial basement membrane regions layers. While IHC localized Tß4 to the leading epithelial edges of the wounds in both groups, staining was more intense in the young corneas. Conclusions: This is the first study to demonstrate the presence of Tß4 in corneas after wounding. In this epithelial wound model, young and aged corneas differ in the rates of healing and in quantity and staining patterns of Tß4. Since 4 is known to promote corneal wound healing, the findings herein may suggest a possible relationship between corneal Tß4 levels and rates of re–epithelialization.

Keywords: cornea: epithelium • wound healing • gene/expression 
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