In vitro Antioxidant and in vivo Wound Healing Activities of the 80% Methanol Extract and Solvent Fractions of Seeds of Brassica carinata A. Braun (Brassicaceae) in Mice

Abstract

Background

Brassica carinata is one of the traditional medicinal plants used in Ethiopia for the treatment of wounds and other diseases. However, the plant has not been scientifically validated, and thus the present study evaluated the in vitro anti-oxidant and the in vivo wound healing activity of the crude extract and solvent fractions of B. carinata seeds in mice.

Materials and Methods

The crude extract was prepared by maceration using 80% methanol and formulated as 5% and 10% w/w ointments for topical application. The acute dermal toxicity was performed in female albino rats based on Organization for Economic Cooperation and Development (OECD) guideline number 434. Excision and incision wound healing models were used to evaluate the wound healing activities of crude extract and solvent fractions ointments in mice. Wound healing parameters such as wound area contraction and the period of epithelialization were determined in an excision model, whereas tensile strength was determined in an incision model. Moreover, the crude extract and solvent fractions were evaluated for the free radical scavenging activities in DPPH assay.

Results

The acute dermal toxicity test showed that a limit dose of 2,000 mg/kg of 10% w/w crude extract ointment did not cause dermal toxicity in mice. In the excision wound model, the data revealed that 10% w/w ointment exhibited a significant wound contraction (from day 6 to 16, P<0.001) effect with a significant decrease in epithelization period (at day 14, P<0.001). In addition, 5% ointment of the crude extract showed a significant effect in wound contraction (from day 8 onwards, P<0.01) and epithelization period (at day 16, P<0.01). Despite all fractions being shown to promote wound healing, 10% w/w aqueous and ethyl acetate fractions showed a significant wound contraction (P<0.001) effect starting from the 4th day onwards. Besides, the maximum antioxidant activity was seen in the aqueous fraction with an IC50 value of 3.45±0.12 mg/mL.

Conclusion

The present study demonstrated that the 80% methanol extract and solvent fractions of the seeds of B.carinata possess potential wound healing and anti-oxidant effects, supporting the traditional use of the plant for wound management.

Keywords:

• wound
• epithelialization
• tensile strength
• incision
• excision
• B. carinata
• antioxidant

Abbreviations

ANOVA, one way analysis of variance; DPPH, 2, 2-diphenyl-2-picrylhydrazyl hydrate; OECD, Organization for Economic Cooperation and Development; SEM, standard error of the mean; SPSS, Statistical Package for Social Sciences.

Data Sharing Statement

All data generated or analyzed during this study are included in the manuscript and are also available from the corresponding author upon request.

Ethics Approval and Consent to Participate

All the experiments were conducted in accordance with a guide for the care and use of laboratory animals: Eighth Edition by National Research Council, Division on Earth and Life Studies, Institute for Laboratory Animal Research, Committee for the Update of the Guide for the Care and Use of Laboratory Animals. We also followed OECD guideline 434. Ethical approval was obtained from Research and Ethical Review Board of College of Medicine and Health Sciences, University of Gondar before the initiation of this study (protocol number sop4/73/09). At the end of the experiment, the animals were killed by Phenobarbital sodium at a dose of 150 mg/kg (euthanasia, mercy killing).

Acknowledgment

The authors would like to thank University of Gondar for the financial support extended to complete the research work.

Author Contributions

All authors made substantial contributions to the conception and design, acquisition of data, or analysis and interpretation of data; took part in drafting the article or revising it critically for important intellectual content; agreed to submit to the current journal; gave final approval of the version to be published; and agree to be accountable for all aspects of the work.

Disclosure

The authors declare that they have no competing interests in this work.

Additional information

Funding

University of Gondar covered the costs of chemicals, reagents and equipments used.