Abstract
Purpose: :
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.
Methods: :
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.
Results: :
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.
Conclusions: :
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.
Keywords: neovascularization • wound healing • inflammation