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N. Fujita, Y. Okada, S. Fujita, K. Fujita, T. Uede, S. Saika; Role of Osteopontin in Vascular Endothelial Canalization in vitro . Invest. Ophthalmol. Vis. Sci. 2009;50(13):4955.
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© ARVO (1962-2015); The Authors (2016-present)
To examine the effect of osteopontin (OPN), a multi-potential extracellular matrix component, for cultured vascular endothelial cells. We previously reported that lacking OPN retarded cauterization-induced neovascularization in cornea in mice (Fujita N, et al. 2008 ARVO). OPN modulates inflammation and arterialization as a ligand of alfa 9 integrin.
Human vascular endothelial cells (HUVECs) were cultured on a fibroblast-feeder layer in 11 days. Synthetic OPN-derived peptide SVVYGLR (0.02, 0.2 and 2.0 ng/ml) or anti-OPN neutralizing antibody (10 µg/ml) developed by us was added to the medium of co-culture. Tube tissue was detected by an immunostaining for PECAM/CD31. The number of the bifurcation and length per one lumen were evaluated.
HUVECs form a vessel-like tube structure when co-cultured with fibroblasts for 11days. Adding peptide SVVYGLR (0.02, 0.2 and 2.0 ng/ml) promoted the canalization and adding anti-OPN neutralizing antibody (10 µg/ml) depressed the canalization.
The present experiments showed that endogenous OPN play a significant role in the neovascularization of cultured cells. The results account for the impaired corneal neovascularization.
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