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

Abstract

Objectives 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 in vitro.Methods 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 0 -azobis(2-amidinopropane) dihydrochloride (AAPH)-induced oxidation and mediate cell migration after artificial wound generation in C2C12 myoblast cells using the scratch wound assay. Key findings All extracts indicated negligible cytotoxicity (IC 50 > 100 lg/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 100 lg/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). Conclusions These findings indicate that the hexane, methanol and water extracts of B. diffusa whole plant promote artificial wound healing and protection against oxidation in vitro and therefore warrant further research into its mechanisms of wound healing.

Effect of ethnomedicinal extracts used for wound healing on cellular migration and intracellular reactive oxygen species release in SC-1 fibroblasts

“…All three plants have been shown to possess a diverse array of phytochemicals with multiple pharmacological properties to attest for their ethnomedicinal use (Okoli et al, 2007;Kuete et al, 2014;Patil and Bhalsing, 2016). In a previous study, the wound healing potential of fractions sequentially extracted from B. diffusa was demonstrated (Yahaya et al, 2018). The current study determined the ability of aqueous ethnomedicinal plant extracts prepared from Aspilia africana, Boerhavia diffusa and Erythrina senegalensis to effect fibroblast migration and ROS release.…”

“…The presence of major phytochemical classes in the extracts was determined using thin layer chromatography (Stahl, 1962). Ultra-performance liquid chromatography, coupled to time of flight mass spectrometry (UPLC-TOF-MS), was employed to verify marker compounds in the extracts (Yahaya et al, 2018). UPLC-TOF-MS was conducted on a Waters instrument coupled in tandem to a Waters photodiode array (PDA) detector.…”

Downregulated miR-204 Promotes Skeletal Muscle Regeneration

“…Scratch Test. The C2C12 myoblasts were manually scraped off with a p200 pipette tip to create scratches [20,21]. After the scratch defect was made, the cells were washed in PBS and cultured in DMEM.…”

AIM: A Computational Tool for the Automatic Quantification of Scratch Wound Healing Assays

“…It consists in using the selection tool of the image analysis software to approximate the wound with a rectangle and then measuring its area [8,9].…”

“…These methods use regular polygons [8,9] to approximate the cell-free area or straight lines [10,11] to estimate the gap width. As a consequence, their accuracy decreases with the irregularity of the wound's edges, and the results are heavily dependent on the operator and his/her ability of consistently evaluate all the images.…”