Investigation of “Wathupalu” (mikania cordata) plant extracts on wound healing using scratch assay

Abstract

Mikania cordata (Brum.) Robinson is a fast-growing perennial vine that belongs to the family of Asteraceae. Different parts of M. cordata are frequently used to treat cuts and wounds in traditional medicine. The aim of this study was to evaluate the wound-healing ability of different extracts of M. cordata leaves and separate the chemical constituents which are responsible for wound healing. Leaf extracts were prepared by sonicating dried M. cordata leaf powder in Dichloromethane (DCM), ethanol, and methanol. Each of these extracts was analyzed on Thin Layer Chromatography (TLC). The scratch assay was performed to analyze the collective cell migration during the wound healing process using the DCM extract on MCF7 epithelial cells. Cells were grown in culture dishes and the wound was created as a scratch when the cells were at 100% confluency. Cells were treated with different concentrations of 100 µg/ml, 500 µg/ml, and 1000 µg/ml of crude DCM extract dissolved in 2.5% (v/v) methanol. The “width of the scratch” was considered as an indicator of collective cell migration. Images were captured at different time intervals of the 0th, 6th and 24th hours during a 24-hour incubation period using an inverted microscope. The width of the scratches was compared with the control {methanol [2.5% (v/v)]}. TLC analysis of ethanol, methanol, and DCM extracts of M. cordata leaves showed that the DCM extract possesses the highest number of chemical components compared to the ethanol and methanol extracts. The results of the scratch assay showed that the DCM extract of M. cordata did not possess a positive effect on collective cell migration within the range of concentrations used during the 24-hour incubation period and methanol caused no effect for Michigan Cancer Foundation-7(MCF7) cells. Further studies with a wide range of concentrations of DCM extract need to be carried out to identify the chemical constituents responsible for scratch closure activity in MCF7 epithelial cells.