Attenuation of oxidative stress and artificial wound closure in C2C12 myoblasts induced by sequential extracts of Boerhavia diffusa.

Abstract

Whole plants of Boerhavia diffusa L. are widely used medicine in Ghana and other tropical countries, for the treatment of wounds and other ailments. The aim of the study was to determine the ability of sequential extracts of B.diffusa to influence oxidation and wound closure in myoblast cells invitro. Sequential extracts were prepared from the whole plant using four solvents of increasing polarity (hexane, ethyl acetate, methanol and water). Cytotoxicity was determined using the sulforhodamine B staining assay, phase-contrast microscopy, plasDIC microscopy and live-dead staining. Extracts were tested for their ability to reduce 2,2'-azobis(2-amidinopropane) dihydrochloride (AAPH)-induced oxidation and mediate cell migration after artificial wound generation in C2C12 myoblast cells using the scratch wound assay. All extracts indicated negligible cytotoxicity (IC50 >100μg/ml), and microscopic evaluation showed no difference from negative controls. AAPH induced a 2.87-fold increase in reactive oxygen species compared to the negative control. Pretreatment with 100μg/ml of the extracts reduced AAPH-induced oxidation to 1.70-fold of the untreated controls (P<0.001). Wound closures in the methanol and water extract treatments were 18.08% and 20.76% higher than the negative control, respectively (P<0.01). These findings indicate that the hexane, methanol and water extracts of B.diffusa whole plant promote artificial wound healing and protection against oxidation invitro and therefore warrant further research into its mechanisms of wound healing.