Vanadate affects proliferation of healing fibroblasts, cellular production of collagen, and expression of alpha-SMA in vitro.
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 alpha-smooth muscle actin (alpha-SMA) production in healing fibroblasts in rat (Sprague-Dawley).


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.
Jiang
Zhi-Tao
ZT
Department of Pediatric Surgery, Second Affiliated Hospital of Harbin Medical University, Harbin, 150086, China.
Li
Zhao-Zhu
ZZ
Jiang
Da-Peng
DP
D000199:Actins / Q000378:metabolism*; D000818:Animals; D049109:Cell Proliferation / Q000187:drug effects*; D003094:Collagen / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D005347:Fibroblasts / Q000187:drug effects*; D005786:Gene Expression Regulation; D066298:In Vitro Techniques; D008022:Ligaments / Q000473:pathology; D008297:Male; D009130:Muscle, Smooth / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D013778:Tetrazolium Salts / Q000494:pharmacology; D013844:Thiazoles / Q000494:pharmacology; D014638:Vanadates / Q000494:pharmacology*
D000199:Actins; D013778:Tetrazolium Salts; D013844:Thiazoles; D014638:Vanadates; D003094:Collagen; C022616:thiazolyl blue