The mechanism of Ficus deltoidea extract in affecting the healing of chronic diabetic wound in vitro

Abstract

Non-healing wound is a serious complication of diabetes mellitus. Untreated chronic diabetic wound negatively impacts human life and increases the risk of mortality and morbidity. F. deltoidea, locally known as Mas Cotek in Malaysia, has been recognised as a significant source of antioxidant, anti-inflammatory, and anti-bacterial components. The current study was designed to evaluate the extract and fractions of F. deltoidea for cytotoxicity and wound healing activity under normal and hyperglycaemic conditions via in vitro study on Human Fibroblast Cells (HSF1184). The extraction and fractionation process was carried out on F. deltoidea dried leaves to obtain the ethanolic extract, chloroform and aqueous fraction. Then, the MTT assay was conducted to determine the cytotoxicity effect of the extract and fractions on HSF1184 cells. Meanwhile, the scratch assay was performed to investigate the wound healing activities for the different concentrations of the F. deltoidea extract and fractions. The MTT assay results revealed that the cytotoxicity was increased in a dose-dependent manner. The IC50 value was recorded for ethanolic extract (144.54 µg/mL), chloroform fraction (128.82 µg/mL) and aqueous fraction (162.18 µg/mL) under normal conditions. The IC50 value for the cells treated under hyperglycaemic conditions was observed at (138.03 µg/mL, 120.22 µg/mL 158.48 µg/mL) in an orderly manner for the three samples. The chloroform fraction was more toxic based on the IC50 value amongst all the treated samples in both conditions. Nonetheless, the results obtained from scratch assay showed that the ethanolic extract (80 µg/mL) and aqueous fraction (40 µg/mL) exerted their stimulatory effects on HSF1184 cells and significantly (*p < 0.05), (**p < 0.01) increased the cell migration rate at 18, 20, 22 and 24 hours in normal condition. The same concentrations for ethanolic extract and aqueous fraction were also statistically significant (*p < 0.05), (**p < 0.01) in terms of HSF1184 cell migration rate under the hyperglycaemic conditions at 22 and 24 hours. However, compared to normal conditions, the cell migration rate was slower under the hyperglycaemic microenvironment as the wound gaps were not fully closed after 24 hours. However, when compared to its control group, the treated group resulted in better migration activity. Meanwhile, the chloroform fraction has no stimulatory effect against HSF1184 in terms of cell migration under normal hyperglycaemic conditions. Taken together, from the findings it is concluded that the ethanolic extract and aqueous fraction significantly exerted their wound healing effect and significantly induced wound healing under normal and hyperglycaemic conditions. Therefore, the ethanolic extract and aqueous fractions need to investigate for further in vitro wound healing studies on other cell line such as keratinocytes and also in vivo animal model.