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Exp Eye Res. 2006 Sep;83(3):502-7. Epub 2006 Apr 21.

Thymosin beta4 inhibits benzalkonium chloride-mediated apoptosis in corneal and conjunctival epithelial cells in vitro.

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Department of Anatomy and Cell Biology, Wayne State University School of Medicine, Scott Hall 8314, 540 E. Canfield Avenue, Detroit, MI 48201, USA.


Thymosin beta-4 (Tbeta(4)) is known to promote ocular wound healing, to decrease ocular inflammation, and to have anti-apoptotic effects on corneal epithelium. In this study, the effect of Tbeta(4) on the survival of human ocular surface epithelial cells exposed to benzalkonium chloride (BAK) was measured. Human conjunctival epithelial cells (HC0597) or human corneal epithelial cells (HCET) were treated with 0%, 0.001%, 0.01%, or 0.1% BAK for 15 min. After 3 or 24h of recovery in culture medium containing 1 microg/ml Tbeta(4), a dosage that has been demonstrated effective in several published studies, DNA synthesis was measured using a colorimetric BrdU incorporation assay. Both conjunctival and corneal epithelial DNA synthesis was inhibited by BAK in a dose-dependent manner. Tbeta(4) did not protect the epithelial cells from BAK-induced inhibition of proliferation. To assess the ability of Tbeta(4) to prevent apoptosis, epithelial cells were treated with 0.01% BAK+Tbeta(4) and cell death was measured using a colorimetric assay. BAK-induced apoptosis increased throughout the duration of the assay, which was carried out to 5 days in culture. Treatment of HC0597 cells with Tbeta(4) significantly inhibited the apoptosis shown to be initiated by BAK. Treatment of non-transformed human corneal epithelial cells (HCEC) with Tbeta(4) also significantly inhibited the apoptosis shown to be initiated by BAK at later times in culture. Ocular solutions containing BAK as a preservative are typically used for extended periods of time. This study suggests that Tbeta(4) may be able to overcome the apoptotic side effect of BAK, and may be a useful additive to solutions containing this preservative.

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