Abstract

Background: The medial collateral ligament of the knee is frequently injured in sports. The medial collateral ligament healing is slow. Vanadate is a transition element that has been shown to be a nonspecific inhibitor of protein tyrosine phosphatases. It has notable effects on wound healing. This study sought to examine the effect of vanadate on proliferation, collagen type I, and α-smooth muscle actin (alpha-SMA) production in healing fibroblasts in rat (Sprague-Dawley).
Material/Methods: Fibroblasts were obtained from a healing medial collateral ligament. Cell cultures were treated with different concentrations of vanadate (5, 10, 20 micromol/L). Healing fibroblasts without vanadate treatment were used control group. Fibroblast proliferation was measured using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium (MTT) assay. Production of collagen type I in a supernatant culture was assayed by enzyme-linked immunosorbent assay (ELISA). Expression of alpha-SMA was assessed by Western blot.
Results: At concentrations of 5 micromol/L, 10 micromol/L, 20 micromol/L of vanadate, fibroblast proliferation was significantly increased. Treatment with vanadate significantly increased the production of collagen type I and decreased a-SMA expression in healing fibroblasts in a dose-dependent manner.
Conclusions: Our results showed that vanadate therapy improves ligament repair via increase in proliferation and collagen type I production, and a decrease in alpha-SMA expression in healing fibroblasts. The results provide a cellular and molecular basis for vanadate’s action as a therapeutic agent for enhancing medial collateral ligament healing and reducing scar formation.

Keywords: Collagen, fibroblast proliferation, vanadate, alpha-smooth muscle actin, Medial collateral ligament, Myofibroblast, Cell Proliferation - drug effects, Collagen - metabolism, Dose-Response Relationship, Drug, Enzyme-Linked Immunosorbent Assay - methods, Fibroblasts - drug effects, Gene Expression Regulation, Ligaments - pathology, Muscle, Smooth - metabolism, Tetrazolium Salts - pharmacology, Thiazoles - pharmacology, Vanadates - pharmacology