Effect Of Anti-Vegf And Anti-Tgfβ molecules In A Rat Wound-healing Model

Abstract

Purpose: : The goal of this study was to test the functional role of a commercial anti-VEGF (Ranibizumab) and anti-TGFβ molecules (P17 and P144 peptides) in wound closure in a rat cell culture wound healing assay.

Methods: : Rat2 fibroblasts were cultured in supplemented serum media (10% bovine serum). Cell migration was assessed by scratch-wounding and the measured area was analysed by Image J software at 24 and 41 hours. Treatment groups were: (A group) serum free media (negative control), (B group) serum media (positive control), (C,D and E groups) Ranibizumab (0.5, 1.0 and 1.5 mg/ml respectively), (F,G and H groups) P17 (100, 150 and 200 mg/ml respectively) and (I and J groups) P144 (50 and 100 mg/ml respectively). B to J groups treatments were performed under medium supplemented with 0.25% rat serum. Statistical analysis was calculated with non-parametrical Kruskal Wallis test and U de Mann-Whitney for groups comparison in SPSS 15.0 software.

Results: : Twenty-four hours after treatment, wound closure area showed a statistically significant reduction in Ranibizumab 1.0 and 1.5 mg/ml (D and E groups; p<0.05) and in P17 100 mg/ml (F group; p<0.05) compared to positive control. Area measurements 41 hours after treatment showed a similar statistical differences in anti-VEGF treatment groups (D, E groups; p<0.05) meanwhile anti-TGFβ treatments only P17 200 mg/ml (H group; p<0.05) showed statistical reduction.

Conclusions: : Ranibizumab and P17 were found effective to inhibit wound closure in rat cultured cells. These results suggest that not only VEGF but also TGFβ inhibition could be an important target in wound healing processes such as corneal and choroidal neovascularization.

Keywords: wound healing • growth factors/growth factor receptors