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S. Presley, A.L. Ripley, M. Osler, D.M. Bader, M.S. Chang; Alterations in a wound healing model by disrupting Bves, an adhesion molecule. . Invest. Ophthalmol. Vis. Sci. 2004;45(13):4900.
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© ARVO (1962-2015); The Authors (2016-present)
Purpose: Bves (blood vessel epicardial substance), is a novel adhesion molecule cloned from a subtraction library enriched for heart–specific gene products, and is believed to be important for cardiac development. Interestingly, we have observed Bves expression in the embryonic and adult eye and in cell culture of human corneal epithelial cells. We hypothesize that Bves is also important for development and maintenance of ocular tissue morphology. This study is initiated to examine the effects of Bves disruption on a cell culture model of corneal wound regeneration and proliferation using synthetic anti–sense morpholine oligonucleotide (GeneTools) against Bves mRNA. Methods: A SV40 immortalized human corneal epithelial cell line is used in cell culture models of cellular proliferation and corneal wounding. Wound assays are performed on cell cultures treated with anti–sense oligo specific to human Bves and a nonspecific control oligo, compared to untreated control cultures. When the culture are at full confluence, a circular wound defect is made in the confluent cells. The defect area is measured daily, using ImageJ software (NIH), from a digital image taken at a fixed magnification. Growth curves are also established for each of the treatment groups. Immunohistiological studies also are carried out using specific primary antibodies to Bves, beta–catenin, E–cadherin, and ZO–1 protiens. Results: In wounding models, the cultures treated with specific anti–sense oligo covered or "healed" the wounded area significantly faster compared to the untreated and control nonspecific oligo. However, treatment with antisense oligo did not affect the growth curve. Immunohistological studies confirmed that Bves expression is decreased with specific Bves anti–sense oligo. Interestingly, regions of decreased Bves treatment are associated with decreased expression of other adhesion molecules. Conclusions: This study demonstrates that disruption of Bves expression in human corneal epithelial cells also leads to disruption of other adhesion molecules at sites of cell–cell contact. The observed physiologic consequence of this interference is a more rapid wound healing in our culture model. However, effects on cellular proliferation are not observed, suggesting alterations in cellular migration.
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