[Effect of vanadate on proliferation and collagen production of medial collateral ligament fibroblasts].
OBJECTIVE
To investigate the effect of vanadate on proliferation and collagen type I production of rat medial collateral ligament (MCL)fibroblasts.


METHODS
A total of 12 adult male SD rats were included, weighing 350-375 g. MCL was cut into small pieces (1 mm x 1 mm x 1 mm) in aseptic conditions, and then placed and cultured in culture chambers. Fibroblasts were passaged with 0.25% trypsin. The vanadate (0, 1.0, 2.5, 5.0 ng/mL) was added in the 3rd passage MCL fibroblasts, respectively, and the samples were divided into 4 groups (0, 1.0, 2.5, 5.0 ng/mL groups). MTT was used to measure the cell proliferation. The production of collagen type I was measured by RT-PCR and ELISA. Twelve samples in each group were measured.


RESULTS
In fibroblast proliferation, the absorbency values of the 1.0, 2.5, 5.0 ng/mL groups were significantly different from that of the 0 ng/mL group (P < 0.05). The absorbency values of the 0, 1.0, 2.5, and 5.0 ng/mL groups were 0.213 +/- 0.016, 0.327 +/- 0.023, 0.449 +/- 0.137, and 0.561 +/- 0.028, respectively. In collagen secretion, vanadate in 1.0, 2.5, 5.0 ng/mL groups could significantly induce the production of collagen type I (P < 0.05) in a dose-dependent manner. The expressions of collagen type I of 0-5 ng/mL groups were 0.47 +/- 0.02, 0.51 +/- 0.03, 0.60 +/- 0.01, and 0.72 +/- 0.02, respectively. There was significant difference between the 1.0, 2.5, 5.0 ng/mL groups and 0 ng mL group (P < 0.05). RT-PCR displayed a dramatic increase of band density. The ratio of band density in the 0-5 ng mL groups was 1.37 +/- 0.76, 1.97 +/- 0.53, 2.41 +/- 0.94, and 2.73 +/- 0.82, respectively. The gene expression of collagen type I in the 1.0, 2.5 and 5.0 ng/mL groups was higher than that in the 0 ng/mL group, and there was significant difference (P < 0.05). There were statistical significant differences among 1.0, 2.5 and 5.0 ng/mL groups in each index mentioned above.


CONCLUSIONS
Vanadate can effectively induce the proliferation of fibroblasts and the production of collagen type I in vitro, which may provide a new approach to the treatment of MCL injury.
Jiang
Dapeng
D
Department of Pediatric Surgery, Second Affiliated Hospital of Harbin Medical University, Harbin Heilongjiang 150086, PR China.
Li
Zhaozhu
Z
Jiang
Zhitao
Z
D000818:Animals; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D003094:Collagen / Q000096:biosynthesis*; D024042:Collagen Type I / Q000096:biosynthesis; D005347:Fibroblasts / Q000187:drug effects* / Q000378:metabolism*; D008297:Male; D017888:Medial Collateral Ligament, Knee / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D014638:Vanadates / Q000494:pharmacology*
D024042:Collagen Type I; D014638:Vanadates; D003094:Collagen