Abstract
Identification and assessment of therapeutic potential of natural products derived from medicinal plants have led to the discovery of innovative and economical drugs to treat several diseases, including chronic wounds. In vitro cell based scratch assay is an appropriate and inexpensive method for initial understanding of wound healing potential of medicinal plant extracts. The current study was aimed at investigating the wound healing capacity of Aristolochia saccata leaf extract by using scratch assay as a primary model, where proliferative and migratory capabilities of test compounds could be monitored through microscopy studies. A. saccata is an evergreen climbing shrub belongs to the family Aristolochiaceae.
Methanolic extraction of the plant material was done using Soxhlet apparatus and the cytotoxicity of the extract on L929 cells was studied by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. L929 is a human fibroblast cell line. In vitro scratch assay was performed to evaluate the wound healing properties of A. saccata leaf extract and possible mechanism of action was analyzed by flow cytometric expression studies of an extracellular matrix (ECM) factor, collagen type-1.
MTT assay revealed that A. saccata leaf extract had no cytotoxic effect on the cells and at higher concentrations, the extract showed mild toxicity resulting in the death of just 2.88% cells. Scratch assay showed 34.05%, 70.00%, 93.52% wound closure at 12hrs, 24hrs and 48hrs of incubation respectively. These results were similar compared to positive control which showed 37.60, 56.41 and 99.05% of wound closure. Further, flow cytometry-based studies revealed that the A. saccata leaf extract induced the expression of ECM remodelling factor collagen-1.
Our study revealed the wound healing capabilities of A. saccata In vitro. Hence, A. saccata could be recommended as a potential source of wound healing agents.