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S. Kanayama, K. Nishida, M. Yamato, R. Hayashi, H. Sugiyama, T. Soma, N. Maeda, T. Okano, Y. Tano; Fibroblast Growth Factor 2 is a Major Angiogenesis Factor in Corneal Vascularization Induced by Cultured Oral Mucosal Epithelial Cell Sheet. Invest. Ophthalmol. Vis. Sci. 2007;48(13):1707. doi: https://doi.org/.
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© ARVO (1962-2015); The Authors (2016-present)
This study was to quantify angiogenesis-induction capability of cultured corneal epithelial cells (CCE) and cultured oral mucosal epithelial cells (COE) in vitro and to investigate a candidate to induce corneal neovasculization after transplantation of cultured oral mucosal epithelial cell sheets.
Rabbit corneal and oral mucosal epithelial cells were cultured with mitomycin-C-treated NIH/3T3 cells on culture plates and culture inserts. After CCE and COE were multilayered, culture medium was replaced by basal medium and incubated. We applied CCE and COE for angiogenesis-induction capability of human umbilical vein endothelial cells (HUVECs) invasion and migration. The protein level of fibroblast growth factor 2 (FGF2), vascular endothelial growth factor (VEGF), Angiopoietin-1 and transforming growth factor ß1 were assessed in conditioned medium by ELISA. Gene expressions of FGF2 and VEGF were determined by real-time PCR. A neutralizing antibody against FGF2 and VEGF were used for blocking HUVECs invasion and migration.
COE induced HUVECs invasion and migration significantly greater compared to CCE. ELISA showed that FGF2 protein level was significantly lower from CCE than COE and that VEGF protein level from CCE was approximately equal to the level from COE. mRNA of FGF2 in COE were expressed significantly more than that in CCE. mRNA of VEGF showed no significant difference between COE and CCE. Anti-FGF2 neutralizing antibody suppressed HUVECs invasion and migration significantly toward COE, whereas no significant suppression was found toward CCE.
Angiogenesis-induction potential toward COE is greater than that toward CCE in vitro. These results suggest FGF2 is a candidate to induce corneal neovasculization after transplantation of cultured oral mucosal epithelial cell sheets.
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