The effect of vascular endothelial growth factor (VEGF) on mouse condylar articular cartilage cultured in vitro.

ABSTRACT

In this study, we explored the direct effect of vascular endothelial growth factor (VEGF) on temporomandibular joint osteoarthritis (TMJ-OA) by analyzing the transformation of mouse condylar cartilage treated in vitro with various concentrations of VEGF. Tissue samples from 126 condyles of four-week-old male C57 mice were randomly divided into 21 groups and treated with VEGF (0 ng/mL, 100 ng/mL, 500 ng/mL, 1 µg/mL, or 2 µg/mL). Furthermore, the samples were treated at different time points (1 d, 2 d, 4 d, and 7 d) and stained with hematoxylin and eosin (HE) and Safranin O and Fast Green stains to observe their morphology. The Mankin score was used to evaluate changes to the condylar cartilage tissues, and immunohistochemistry was performed to observe the expressions of VEGF receptor 2 (VEGFR2), matrix metallopeptidase 9 (MMP9), matrix metallopeptidase 13 (MMP13), and tumor necrosis factor-related activation-induced cytokine (TRANCE). An HE staining analysis revealed that the experimental groups treated with VEGF exhibited the destruction of their condylar cartilage and a proliferation of their hypertrophic cells, in comparison to the control group. Safranin O and Fast Green staining showed that the experimental groups had decreased levels of proteoglycan and degenerative changes in their condylar cartilage. The Mankin score of the samples increased with increasing concentration and treatment time of VEGF, and the differences between the groups were statistically significant (P < 0.05). Immunohistochemistry demonstrated that the expression levels of VEGFR2, MMP9, MMP13, and TRANCE significantly increased in the experimental groups, in comparison to those in the control group, suggesting that VEGF promoted TMJ-OA in mice in vitro.