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Pharmacognosy Magazine ,2019,15,66,483-494.DOI:10.4103/pm.pm_81_19Published:nType:Original Article Authors:Murad Alsawalha, Abeer Mohammed Al-Subaie, Reem Yousuf Al-Jindan, Srinivasa Rao Bolla, Mohammed Salahuddin, Vishnu Priya Veeraraghavan, Dwaipayan Sen, Janardhana Papayya Balakrishna, Padma Kanchi Ravi, Joel Palpath Joseph, Aruthra Arumugam Pillai, Shiva Shankar Reddy Gollapalli, Shonima Pala, Fajre Pengateeri, George Dominic, and Surapaneni Krishna Mohan Author(s) affiliations:Murad Alsawalha1, Abeer Mohammed Al-Subaie2, Reem Yousuf Al-Jindan3, Srinivasa Rao Bolla4, Mohammed Salahuddin5, Vishnu Priya Veeraraghavan6, Dwaipayan Sen7, Janardhana Papayya Balakrishna7, Padma Kanchi Ravi8, Joel Palpath Joseph9, Aruthra Arumugam Pillai9, Shiva Shankar Reddy Gollapalli9, Shonima Pala10, Fajre Pengateeri11, George Dominic12, Surapaneni Krishna Mohan13 1Department of Chemical and Process Engineering Technology, Jubail Industrial College, Saudi Arabia 2Department of Clinical Laboratory Sciences, College of Applied Medical Sciences, Imam Abdulrahman Bin Faisal University, Dammam, Kingdom Saudi Arabia 3Department of Microbiology, Nazarbayev University School of Medicine (NUSOM), Nazarbayev University, Nur-Sultan City, Kazakhstan, Kazakhstan 4Department of Biomedical Sciences, Nazarbayev University School of Medicine (NUSOM), Nazarbayev University, Nur-Sultan City, Kazakhstan, Kazakhstan 5Department of Animal House, Institute of Research and Medical Consultations, Imam Abdulrahman Bin Faisal University, Dammam, Kingdom Saudi Arabia 6Department of Biochemistry, Saveetha Dental College, Saveetha Institute of Medical and Technical Sciences, Saveetha University, Chennai, Tamil Nadu, India 7Centre for Biomaterials, Cellular and Molecular Theranostics, VIT University, Vellore, Tamil Nadu, India 8Department of Biotechnology, Sri Padmavati Mahila Visvavidyalayam, Tirupati, Andhra Pradesh, India 9Department of Biotechnology, Stellixir Biotech Private Ltd., Bengaluru, Karnataka, India 10Central Animal House Facility, Pondicherry University, Puducherry, India 11Department of General Education, Directorate of Vocational and Higher Secondary Education, Government of Kerala, Thiruvananthapuram, Kerala, India 12Dairy Cattle Nutrition Division, ICAR-National Dairy Research Institute, Karnal, Haryana, India 13Department of Medical Biochemistry, College of Applied Medical Sciences in Jubail, Imam Abdulrahman Bin Faisal University, Jubail, Saudi Arabia Abstract:Background:Easy availability, relatively low cost with fewer side effects, has made the herbal extracts/fractions/pure compounds as prominent source of medicinally important molecules. Dioscorea villosa L. commonly known as wild yam belongs to the family Dioscoreaceae and has been used in various parts of India to treat joint pain, arthritis, and various other diseases. However, its role in wound healing has not been documented so far. In the current study, the in vitro wound healing capabilities of D. villosa were examined using L929 cells. Materials and Methods:Methanolic extraction of D. villosa leaves was prepared by applying inexpensive maceration method. 3-(4, 5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay was used to analyze the cytotoxicity of D. villosa extract and in vitro wound healing capabilities were investigated by applying scratch assay. The qualitative measurement of different secondary metabolites was determined by standard biochemical assays. Gas chromatography-mass spectrometry (GC-MS) was performed to identify the possible wound healing components present in the methanolic leaf extract of D. villosa, and the antioxidant properties of the plant extract were evaluated by α,α-diphenyl-β-picrylhydrazyl and ferric reducing antioxidant power assays. Furthermore, the possible molecular factors involved in the proliferation and migration of fibroblast in the presence of D. villosa extract was determined by flow cytometry technique. Results:The experiments to analyze the cytotoxic effect of D. villosa on L929 cells revealed that at the highest concentration used, i.e., 500 μg/mL after 48 h of incubation, 96.06% ± 0.42% of the cells were viable. The results of the scratch assay revealed that 125 μg/mL of plant extract induced the migration in 88.58% of fibroblast cells. Through GC-MS analysis, antioxidant and anti-inflammatory molecules such as 1 H-Indole-2,3-dione (Isatin) and Dexamethasone have been identified. In addition, flow cytometry data showed the influence of plant extract on the expression of Collagen-1 and transforming growth factor (TGF)-beta, which play a major role in the wound healing processes. 125 μg/mL of plant extract induced Collagen-1 in 22.18% cells and TGF-beta in 80.77% of cells, respectively. Conclusion:The presence of potent antioxidant and anti-inflammatory molecules and capability to induce the expression of fundamental wound healing molecular factors TGF-beta and collagen-1 in fibroblast cells, endorsed D. villosa as a potential wound healing agent. Keywords:Collagen-1, Dioscorea villosa, gas chromatography-mass spectrometry, scratch assay, transforming growth factor-beta, Wound healingView:PDF (3.08 MB)