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The Modulating Function of Osteopontin and Osteopontin Monoclonal Antibody in Human Tenon's Fibroblasts |
Weina Li, Jinwei Cheng, Ruili Wei. |
Department of Ophthalmology, Shanghai Changzheng Hospital, Second Military Medical University,Shanghai 200003, China |
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Abstract Objective: To assess the modulating function of osteopontin (OPN) on human Tenon's fibroblasts (HTFs). Methods: With the intervention of OPN in different concentrations (0, 0.05 μmol/L, 0.5 μmol/L,and 5 μmol/L), the proliferation rate of HTFs was measured by 3-(4,5-dimethylthiazol-2-Yl)-2,5-
diphenyltetrazolium bromide (MTT), and the motility was surveyed by the cell scratch method. Real-time polymerase chain reaction (PCR) was applied to measure the mRNA of vascular endothelial growth factor(VEGF) and transforming growth factor β (TGF-β) in HTFs in the presence of different concentration of OPN. Western blot was used to measure the collagen-I levels. Data were analyzed using ANOVA. Results:OPN induced the proliferation of HTFs as indicated by MTT and enhanced the motility of HTFs as shownby the cell scratch method. The effects were concentration dependent (F =174.5, 297.1, P <0.01). In the presence of anti-OPN 1A12 (5 μmol/L OPN+20 μg/ml 1A12), the effects of OPN on HTFs were inhibited. None of the concentrations of OPN affected mRNA levels of VEGF and TGF-β mRNA. OPN augmented HTF expression of collagen-I protein in a concentration-dependent manner (F =30.2, P <0.01). Conclusions:The enhancement of HTF proliferation, motility, and collagen-I expression OPN was concentration dependent. Incubation with the monoclonal OPN antibody 1A12 inhibited the effects of OPN on the HTFs.
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Corresponding Authors:
Jinwei Cheng, Department of Ophthalmology, Shanghai Changzheng Hospital,
Second Military Medical University, Shanghai 20003, China (Email: jinnwave@163.com)
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