June 2015
Volume 56, Issue 7
Free
ARVO Annual Meeting Abstract  |   June 2015
Platelet Recruitment is Associated with Limbal Venule Expansion in a Mouse Model of Corneal Abrasion
Author Affiliations & Notes
  • Sri Magadi
    College of Optometry, University of Houston, Houston, TX
  • Zhijie Li
    Pediatrics-leukocyte biology, Baylor College of Medicine, Houston, TX
  • Wanyu Zhang
    College of Optometry, University of Houston, Houston, TX
  • Debjani Phillips
    University of Wisconsin, Madison, WI
  • Rolando Rumbaut
    Pediatrics-leukocyte biology, Baylor College of Medicine, Houston, TX
    Michael E. DeBakey VA Medical Center, Houston, TX
  • Clifton Wayne Smith
    Pediatrics-leukocyte biology, Baylor College of Medicine, Houston, TX
  • Alan Robert Burns
    College of Optometry, University of Houston, Houston, TX
    Pediatrics-leukocyte biology, Baylor College of Medicine, Houston, TX
  • Footnotes
    Commercial Relationships Sri Magadi, None; Zhijie Li, None; Wanyu Zhang, None; Debjani Phillips, None; Rolando Rumbaut, None; Clifton Smith, None; Alan Burns, None
  • Footnotes
    Support None
Investigative Ophthalmology & Visual Science June 2015, Vol.56, 743. doi:
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      Sri Magadi, Zhijie Li, Wanyu Zhang, Debjani Phillips, Rolando Rumbaut, Clifton Wayne Smith, Alan Robert Burns; Platelet Recruitment is Associated with Limbal Venule Expansion in a Mouse Model of Corneal Abrasion. Invest. Ophthalmol. Vis. Sci. 2015;56(7 ):743.

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

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Abstract

Purpose: A hallmark of acute inflammation is vasodilation. The extravasation of neutrophils and platelets after corneal abrasion is beneficial to wound healing. In mast cell deficient mice (Kitw-sh/w-sh), limbal venule expansion is blunted, there is a marked delay in neutrophil extravasation, and little or no platelet extravasation. The purpose of this study was to examine the relationship between venule expansion and platelet extravasation.

Methods: Adult C57BL/6 mice were anesthetized and a 2mm central corneal abrasion was made using a golf-club spud or Alger brush. Prior to injury, one group of wild type mice was treated with anti-Ly6G antibody to deplete neutrophils which in turn limits platelet recruitment. A second group had reduced leukocyte CD18 expression (CD18 mutant) with normal neutrophil extravasation but limited platelet recruitment. The third group of wild type injured mice received topical treatment with rIL-20 (200 ng/ml) once every 4h for 24h which inhibits neutrophil and platelet extravasation. A control group of injured wild type mice received phosphate buffered saline topically. At 24h post-injury, excised corneas were immunostained for vessels and platelets. Limbal venule diameters were evaluated and analyzed using the Kolmogorov-Smirnov test.

Results: Prior to injury, limbal venule diameters were not different across mouse groups. At 24h post-injury, in wild type mice, the average limbal venule diameters increased from 15 μm to 25 μm (p<0.05 compared to uninjured baseline). Injured CD18 mutant mice and mice treated with anti-Ly6G or rIL-20 showed a marked reduction (>70%) in platelet extravasation compared to injured wild type mice. The average venule diameters in injured CD18 mutant mice and mice treated with anti-Ly6G or rIL-20 were similar across the groups, increasing to ~20 μm which was significantly less than that observed in injured wild type mice.

Conclusions: In three experimental models that significantly reduce platelet extravasation following corneal injury, limbal venule expansion was significantly reduced, suggesting possible functional links between venule expansion and platelet extravasation.

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