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Evaluation of In vitro Wound Healing Activity of Thymoquinone

Year 2020, Volume: 79 Issue: 2, 151 - 156, 25.12.2020

Abstract

Objective: Nigella sativa has been extensively investigated as an important potential agent for the healing of wounds and there have been numerous studies regarding its effect. Although thymoquinone (TQ) is a well-known active constituent of Nigella sativa, studies in to the usability of TQ on wound healing are still insufficient. In this study, we aimed to evaluate the in vitro wound healing potential of TQ. Materials and Methods: NIH/3T3 mouse embryonic fibroblast cells were used to evaluate the wound healing effect of TQ. Different concentrations of TQ (0.1, 1 and 10 μM) were applied to the cells and their cytotoxic effect on cells after 24- and 48- hours was measured by MTT assay. Its effect on wound healing after 18- and 24- hours recovery was examined by in vitro scratch assay. Also, the level of β-catenin, an effective protein in the process of healing wounds, was determined by Western blot assay. Results: MTT analysis indicated that 0.1, 1 and 10 μM doses of TQ had increased the cell numbers. In vitro scratch assay data showed that treatment with 1 and 10 μM TQ resulted in a statistically significant wound closure activity (91.35% and 90.84%, respectively) compared to the control. Additionally, we observed a statistically significant increase in the β-catenin protein level which supported our data. Conclusion: Our results demonstrated that TQ increases both the viability of NIH/3T3 cells and its wound closure activity in vitro, and that it has the effect of increasing crucial protein β-catenin. This study suggests that TQ may be a valuable substance for the healing of wounds and that its usability should be investigated.

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References

  • 1. Ab Rahman MR, Abdul Razak F, Mohd Bakri M. Evaluation of wound closure activity of Nigella sativa, Melastoma malabathricum, Pluchea indica, and Piper sarmentosum extracts on scratched monolayers of human gingival fibroblasts. J Evid Based Complementary Altern Med 2014; 1-9.
  • 2. Mathieu D, editor. Handbook on hyperbaric medicine. New York: Springer; 2006.
  • 3. Menke NB, Ward KR, Witten TM, Bonchev DG, Diegelmann RF. Impaired wound healing. Clin Dermatol 2007; 25: 19–25.
  • 4. Nelzen O, Bergqvist D, Lindhagen A. The prevalence of chronic lower-limb ulceration has been underestimated: Results of a validated population questionnaire. Br J Surg 1996; 83: 255–8.
  • 5. Agyare C, Bekoe EO, Boakye YD, Dapaah SO, Appiah T, Bekoe SO. Medicinal Plants and Natural Products with Demonstrated Wound Healing Properties. Wound Healing: New insights into Ancient Challenges. London: InTech; 2016.
  • 6. Sarkhail P, Esmaily H, Baghaei A, Shafiee A, Abdollahi M, Khademi Y, et al. Burn healing potential of Nigella sativa seed oil in rats. Int J Pharm Sci Res 2011; 2(1): 34-40.
  • 7. Eid AM, Elmarzugi NA, Abu Ayyash LM, Sawafta MN, Daana HI. A Review on the Cosmeceutical and External Applications of Nigella sativa. J Trop Med 2017; 1-6
  • 8. Yaman I, Durmus AS, Ceribasi S, Yaman M. Effects of Nigella sativa and silver sulfadiazine on burn wound healing in rats. Vet Med (Praha) 2010; 55(12): 619-24.
  • 9. Nourbar E, Mirazi N, Yari S, Rafieian-Kopaei M, Nasri H. Effect of hydroethanolic extract of Nigella sativa L. on skin wound healing process in diabetic male rats. Int J Prev Med 2019; 10:18-24.
  • 10. Jagetia GC, Ravikiran P. Acceleration of wound repair and regeneration by Nigella sativa in the deep dermal excision wound of mice whole body exposed to different doses of γ-radiation. Am Res J Med Surg 2015; 1(3): 1-17.
  • 11. Kouidhi B, Zmantar T, Jrah H, Souiden Y, Chaieb K, Mahdouani K et al. Antibacterial and resistance-modifying activities of thymoquinone against oral pathogens. Ann Clin Microbiol Antimicrob 2011; 10: 29-35.
  • 12. Halawani E. Antibacterial Activity of Thymoquinone and Thymohydroquinone of Nigella sativa L. and Their Interaction with Some Antibiotics. Adv Biol Res 3. 2009; (5-6): 148-52.
  • 13. Hayat K, Asim MB, Nawaz M, Li M, Zhang L, Sun N. Ameliorative effect of thymoquinone on ovalbumin-induced allergic conjunctivitis in Balb/c mice. Curr Eye Res 2011; 36(7): 591-8.
  • 14. Abdelmeguid NE, Fakhoury R, Kamal SM, Al Wafai RJ. Effects of Nigella sativa and thymoquinone on biochemical and subcellular changes in pancreatic beta-cells of streptozotocin-induced diabetic rats. J Diabetes 2010; 2(4): 256-66.
  • 15. Ali BH, Blunden G. Pharmacological and toxicological properties of Nigella sativa. Phytother Res 2003; 17(4): 299-305.
  • 16. Hosseinzadeh H, Parvardeh S, Asl MN, Sadeghnia HR, Ziaee T. Effect of thymoquinone and Nigella sativa seeds oil on lipid peroxidation level during global cerebral ischemia-reperfusion injury in rat hippocampus. Phytomedicine 2007; 14(9): 621-7.
  • 17. Şengelen A, Önay-Uçar E. Rosmarinic acid and siRNA combined therapy represses Hsp27 (HSPB1) expression and induces apoptosis in human glioma cells. Cell Stress Chaperones. 2018; 23(5): 885–96.
  • 18. Zedlitz S, Kaufmann R, Boehncke WH. Allergic contact dermatitis from black cumin (Nigella sativa) oil-containing ointment. Contact Dermatitis 2002; 46(3): 188.
  • 19. Gelot P, Bara-Passot C, Gimenez-Arnau E, Beneton N, Maillard H, Celerier P. Éruption Bulleuse À L’Huile De Nigelle. Ann Dermatol Venereol 2012; 139(4): 287–91.
  • 20. Kant V, Gopal A, Pathak NN, Kumar P, Tandan SK, Kumar D. Antioxidant and anti-inflammatory potential of curcumin accelerated the cutaneous wound healing in streptozotocin-induced diabetic rats. Int Immunopharmacol 2014; 20: 322-30.
  • 21. Sayin O, Micili SC, Goker A, Kamaci G, Ergur BU, Yilmaz O et al. The role of resveratrol on full–Thickness uterine wound healing in rats. Taiwan J Obstet Gynecol 2017; 56(5): 657-63.
  • 22. Rocha, J, Eduardo‐Figueira M, Barateiro A, Fernandes A, Brites D, Bronze R et al. Anti‐inflammatory effect of rosmarinic acid and an extract of Rosmarinus officinalis in rat models of local and systemic inflammation. Basic Clin Pharmacol Toxicol 2015; 116(5): 398-413.
  • 23. Selçuk CT, Durgun M, Tekin R, Yolbas L, Bozkurt M, Akçay C et al. Evaluation of the effect of thymoquinone treatment on wound healing in a rat burn model. J Burn Care Res. 2013; 34(5): 274-81.
  • 24. Yusmin A, Ahmad N. Effect of thymoquinone on wound healing on wound healing in alloxan-induced diabetic rats. Asian J Pharm Clin Res 2017; 10(9): 242-5.
  • 25. Liang CC, Park AY, Guan JL. In vitro scratch assay: a convenient and inexpensive method for analysis of cell migration in vitro. Nat Protoc 2007; 2(2): 329-33.
  • 26. Bielefeld Kirsten A, Amini-Nik S, Alman BA. Cutaneous wound healing: recruiting developmental pathways for regeneration. Cell Mol Life Sci 2013; 70(12): 2059-81.
  • 27. Polakis P. Wnt signaling and cancer. Genes Dev 2000; 14: 1837–51.
  • 28. Taipale J, Beachy PA. The hedgehog and Wnt signalling pathways in cancer. Nature 2001; 411: 349–54.
  • 29. Korswagen HC, Clevers HC. Activation and repression of wing-less/Wnt target genes by the TCF/LEF-1 family of transcription factors. Cold Spring Harb Symp Quant Biol 1999; 64: 141–7.
Year 2020, Volume: 79 Issue: 2, 151 - 156, 25.12.2020

Abstract

References

  • 1. Ab Rahman MR, Abdul Razak F, Mohd Bakri M. Evaluation of wound closure activity of Nigella sativa, Melastoma malabathricum, Pluchea indica, and Piper sarmentosum extracts on scratched monolayers of human gingival fibroblasts. J Evid Based Complementary Altern Med 2014; 1-9.
  • 2. Mathieu D, editor. Handbook on hyperbaric medicine. New York: Springer; 2006.
  • 3. Menke NB, Ward KR, Witten TM, Bonchev DG, Diegelmann RF. Impaired wound healing. Clin Dermatol 2007; 25: 19–25.
  • 4. Nelzen O, Bergqvist D, Lindhagen A. The prevalence of chronic lower-limb ulceration has been underestimated: Results of a validated population questionnaire. Br J Surg 1996; 83: 255–8.
  • 5. Agyare C, Bekoe EO, Boakye YD, Dapaah SO, Appiah T, Bekoe SO. Medicinal Plants and Natural Products with Demonstrated Wound Healing Properties. Wound Healing: New insights into Ancient Challenges. London: InTech; 2016.
  • 6. Sarkhail P, Esmaily H, Baghaei A, Shafiee A, Abdollahi M, Khademi Y, et al. Burn healing potential of Nigella sativa seed oil in rats. Int J Pharm Sci Res 2011; 2(1): 34-40.
  • 7. Eid AM, Elmarzugi NA, Abu Ayyash LM, Sawafta MN, Daana HI. A Review on the Cosmeceutical and External Applications of Nigella sativa. J Trop Med 2017; 1-6
  • 8. Yaman I, Durmus AS, Ceribasi S, Yaman M. Effects of Nigella sativa and silver sulfadiazine on burn wound healing in rats. Vet Med (Praha) 2010; 55(12): 619-24.
  • 9. Nourbar E, Mirazi N, Yari S, Rafieian-Kopaei M, Nasri H. Effect of hydroethanolic extract of Nigella sativa L. on skin wound healing process in diabetic male rats. Int J Prev Med 2019; 10:18-24.
  • 10. Jagetia GC, Ravikiran P. Acceleration of wound repair and regeneration by Nigella sativa in the deep dermal excision wound of mice whole body exposed to different doses of γ-radiation. Am Res J Med Surg 2015; 1(3): 1-17.
  • 11. Kouidhi B, Zmantar T, Jrah H, Souiden Y, Chaieb K, Mahdouani K et al. Antibacterial and resistance-modifying activities of thymoquinone against oral pathogens. Ann Clin Microbiol Antimicrob 2011; 10: 29-35.
  • 12. Halawani E. Antibacterial Activity of Thymoquinone and Thymohydroquinone of Nigella sativa L. and Their Interaction with Some Antibiotics. Adv Biol Res 3. 2009; (5-6): 148-52.
  • 13. Hayat K, Asim MB, Nawaz M, Li M, Zhang L, Sun N. Ameliorative effect of thymoquinone on ovalbumin-induced allergic conjunctivitis in Balb/c mice. Curr Eye Res 2011; 36(7): 591-8.
  • 14. Abdelmeguid NE, Fakhoury R, Kamal SM, Al Wafai RJ. Effects of Nigella sativa and thymoquinone on biochemical and subcellular changes in pancreatic beta-cells of streptozotocin-induced diabetic rats. J Diabetes 2010; 2(4): 256-66.
  • 15. Ali BH, Blunden G. Pharmacological and toxicological properties of Nigella sativa. Phytother Res 2003; 17(4): 299-305.
  • 16. Hosseinzadeh H, Parvardeh S, Asl MN, Sadeghnia HR, Ziaee T. Effect of thymoquinone and Nigella sativa seeds oil on lipid peroxidation level during global cerebral ischemia-reperfusion injury in rat hippocampus. Phytomedicine 2007; 14(9): 621-7.
  • 17. Şengelen A, Önay-Uçar E. Rosmarinic acid and siRNA combined therapy represses Hsp27 (HSPB1) expression and induces apoptosis in human glioma cells. Cell Stress Chaperones. 2018; 23(5): 885–96.
  • 18. Zedlitz S, Kaufmann R, Boehncke WH. Allergic contact dermatitis from black cumin (Nigella sativa) oil-containing ointment. Contact Dermatitis 2002; 46(3): 188.
  • 19. Gelot P, Bara-Passot C, Gimenez-Arnau E, Beneton N, Maillard H, Celerier P. Éruption Bulleuse À L’Huile De Nigelle. Ann Dermatol Venereol 2012; 139(4): 287–91.
  • 20. Kant V, Gopal A, Pathak NN, Kumar P, Tandan SK, Kumar D. Antioxidant and anti-inflammatory potential of curcumin accelerated the cutaneous wound healing in streptozotocin-induced diabetic rats. Int Immunopharmacol 2014; 20: 322-30.
  • 21. Sayin O, Micili SC, Goker A, Kamaci G, Ergur BU, Yilmaz O et al. The role of resveratrol on full–Thickness uterine wound healing in rats. Taiwan J Obstet Gynecol 2017; 56(5): 657-63.
  • 22. Rocha, J, Eduardo‐Figueira M, Barateiro A, Fernandes A, Brites D, Bronze R et al. Anti‐inflammatory effect of rosmarinic acid and an extract of Rosmarinus officinalis in rat models of local and systemic inflammation. Basic Clin Pharmacol Toxicol 2015; 116(5): 398-413.
  • 23. Selçuk CT, Durgun M, Tekin R, Yolbas L, Bozkurt M, Akçay C et al. Evaluation of the effect of thymoquinone treatment on wound healing in a rat burn model. J Burn Care Res. 2013; 34(5): 274-81.
  • 24. Yusmin A, Ahmad N. Effect of thymoquinone on wound healing on wound healing in alloxan-induced diabetic rats. Asian J Pharm Clin Res 2017; 10(9): 242-5.
  • 25. Liang CC, Park AY, Guan JL. In vitro scratch assay: a convenient and inexpensive method for analysis of cell migration in vitro. Nat Protoc 2007; 2(2): 329-33.
  • 26. Bielefeld Kirsten A, Amini-Nik S, Alman BA. Cutaneous wound healing: recruiting developmental pathways for regeneration. Cell Mol Life Sci 2013; 70(12): 2059-81.
  • 27. Polakis P. Wnt signaling and cancer. Genes Dev 2000; 14: 1837–51.
  • 28. Taipale J, Beachy PA. The hedgehog and Wnt signalling pathways in cancer. Nature 2001; 411: 349–54.
  • 29. Korswagen HC, Clevers HC. Activation and repression of wing-less/Wnt target genes by the TCF/LEF-1 family of transcription factors. Cold Spring Harb Symp Quant Biol 1999; 64: 141–7.
There are 29 citations in total.

Details

Primary Language English
Journal Section Research Articles
Authors

Murat Pekmez This is me 0000-0002-6150-8372

Nihan Sultan Mılat This is me 0000-0002-0795-8030

Publication Date December 25, 2020
Submission Date January 2, 2020
Published in Issue Year 2020 Volume: 79 Issue: 2

Cite

AMA Pekmez M, Mılat NS. Evaluation of In vitro Wound Healing Activity of Thymoquinone. Eur J Biol. December 2020;79(2):151-156.