In vitro wound healing potency of methanolic leaf extract of Aristolochia saccata is possibly mediated by its stimulatory effect on collagen-1 expression

Srinivasa Rao Bolla; Abeer Mohammed Al-Subaie; Reem Yousuf Al-Jindan; Janardhana Papayya Balakrishna; Padma Kanchi Ravi; Vishnu Priya Veeraraghavan; Aruthra Arumugam Pillai; Shiva Shankar Reddy Gollapalli; Joel Palpath Joseph; Krishna Mohan Surapaneni

doi:10.1016/j.heliyon.2019.e01648

In vitro wound healing potency of methanolic leaf extract of Aristolochia saccata is possibly mediated by its stimulatory effect on collagen-1 expression

Heliyon. 2019 May 20;5(5):e01648. doi: 10.1016/j.heliyon.2019.e01648. eCollection 2019 May.

Affiliations

¹ Department of Anatomy, College of Medicine, Imam Abdulrahman Bin Faisal University, P.O. Box 2114, Dammam 31451, Saudi Arabia.
² Department of Clinical Laboratory Sciences, College of Applied Medical Sciences, Imam Abdulrahman Bin Faisal University, P.O. Box 2435, Dammam 31451, Saudi Arabia.
³ Department of Microbiology, College of Medicine, Imam Abdulrahman Bin Faisal University, P.O. Box 2114, Dammam 31451, Saudi Arabia.
⁴ Department of Biotechnology, Stellixir Biotech Private Ltd, Peenya 2nd Stage Industrial Area, Bangalore 560058, Karnataka, India.
⁵ Department of Biotechnology, Sri Padmavati Mahila Visva Vidyalayam University, Tirupati, Andhra Pradesh, India.
⁶ Department of Biochemistry, Saveetha Dental College, Saveetha Institute of Medical and Technical Sciences (SIMATS), Saveetha University, 162, P. H. Road, Velappanchavadi, Chennai 600 077, Tamil Nadu, India.
⁷ Department of Medical Biochemistry, College of Applied Medical Sciences - Jubail (CAMSJ), Imam Abdulrahman Bin Faisal University, P.O. Box 4030, Al Ansar Rd, Deffi, Jubail Industrial City, Al Jubail 35816, Saudi Arabia.

Abstract

Background: 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.

Methods: 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.

Results: 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.

Conclusion: 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.

Keywords: Biochemistry; Biotechnology; Cell biology; Molecular biology; Pharmaceutical science; Plant biology.