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In vitro Investigation of Wound Healing, Cytotoxic, Antioxidant Properties of Shed Snake Skin (Dolichophis caspius) Aqueous Extract

Year 2026, Volume: 16 Issue: 1, 244 - 255, 23.01.2026
https://doi.org/10.31020/mutftd.1749015

Abstract

Purpose: As cold-blooded reptiles, snakes seasonally shed dead skin cells due to thickening of the skin and removing harmful parasites during growth. Dolichophis caspius is a popularly known snake in Türkiye that sheds its skin at certain times of the year. The focus of this study was to determine the total phenolic and total flavonoid contents, as well as the antioxidant activity, of the aqueous extracts of D. caspius shed skin and to investigate the effects of these extracts on cytotoxicity and wound healing.
Method: The shed snake skins were extracted using distilled water. Then, the total phenolic contents, the total flavonoid contents, and the antioxidant activity were evaluated through the Folin-Ciocalteu method, the aluminum-chloride colorimetric assay, and the DPPH method, respectively. The cytotoxic activity was determined by MTT assay and the effect on in vitro wound healing was determined by scratch assay.
Findings: Total phenolic content, total flavonoid content, and antioxidant activity of the extract were calculated as 681±46 µg GAE/g, 442.35±13 µg CE/g, and 13.06%, respectively. IC50 values were determined as 2972±179 µg/mL and 2657±128 µg/mL for dermal fibroblast cells (BJ cell line) and melanoma cells (SK-MEL-30 cell line) in 72 hours. In comparison to the control group, it was found that the extracts at the studied doses do not have effective wound closure rates in BJ cells.
Conclusion: It has been determined that shed snake skins will have promising results in cytotoxicity and wound healing studies thanks to their potentially active biomolecules.

Supporting Institution

This study was supported by the TUBITAK 2209-A University Students Research Projects with application number 1919B012325139.

Project Number

1919B012325139

Thanks

The authors would like to thank Prof. Dr. Hikmet Sami Yıldırımhan, a faculty member in the Zoology Department of the Faculty of Arts and Sciences at Bursa Uludağ University, for his work on identifying the species of shed snake skins.

References

  • 1. Iowa [Internet] Department of Natural Resource. Why do snakes shed their skin? [updated 2023 Nov 02; cited 2025 Jul 23]. Available from: https://www.iowadnr.gov/About-DNR/DNR-News-Releases/ArticleID/158/Why-Do-Snakes-Shed-Their-Skin
  • 2. Sakalli V, et al. Investigation of the usability of snake shirts in the detection of morphological differences between snake species. Eskisehir Technical University Science and Technology Journal-C Life Sciences and Biotechnology 2020;9(1):26-34.
  • 3. Sinmez C, et al. Investigation of immunomodulatory and cytotoxic effects of shed snake skin (Elaphe sauromates) extract. Front.pharmacol. 2024;15,1270970.
  • 4. Andonov K, et al. Analyses of skin secretions of Vipera ammodytes (Linnaeus, 1758) (Reptilia: Serpentes), with focus on the complex compounds and their possible role in the chemical communication. Molecules 2020;25(16):3622.
  • 5. Demı̇rcı̇ ÜÖ. Snake in historical dialects. Electronic Turkish Studies 2014;9(5).
  • 6. Dönmez E. Oral snake skı̇n resultı̇ng in anaphylaxı̇s: How and why? J Emerg Med Case Rep 2020;11(4):95–97.
  • 7. Sakalli V, Çömelekoğlu Ü. Determination of the elemental content of snake shirts used in the treatment of warts in Anatolian folkloric medicine. Mersin University Faculty of Medicine Lokman Hekim Journal of History of Medicine and Folkloric Medicine 2019;9(3):312-316.
  • 8. Khalili F. Studies on the morphology of Dolichophis caspius (Gmeli̇n, 1789) (Serpentes: Colubridae) distributed in western Anatolia, Ege University Biology Department Master Thesis, 2018.
  • 9. Marcov SDC, David A. Dolichophis caspius (Serpentes: Colubridae) in Romania: New distribution records from the northern limit of its range. Turk J Zool 2010:34(1):119-121.
  • 10. Pourmorad F, Hosseinimehr SJ, Shahabimajd N. Antioxidant activity, phenol and flavonoid contents of some selected Iranian medicinal plants. Afr J Biotechnol 2006;5(11).
  • 11. Adam M, Rasad MSBA. Expression of matrix metalloproteinase-13 in human skin melanoma cancer treated by Baccaurea angulata in vitro. J basic appl Res Biomed 2025;1(1):21-28.
  • 12. Yetişgi̇n A. Effects of nanomaterials on stem cell fate determination. Yıldız Technical University Bioengineering Department Graduation Thesis 2018.
  • 13. Guo S, DiPietro LA. Factors affecting wound healing. J Dent Res 2010;89(3):219–229.
  • 14. Gonzalez AC, et al. Wound healing - A literature review. Anais Brasileiros de Dermatologia 2016;91(5):614–620.
  • 15. Öztopalan DF, Recep I, Durmmuş AS. The role of growth factors and cytokines in wound healing. Journal of Dicle University Veterinary Faculty 2017;10(1):83-88.
  • 16. Stamm A, et al. In vitro wound healing assays - state of the art. BioNanoMaterials 2016;17(1-2):79-87.
  • 17. 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):329–333.
  • 18. Kupina S, et al. Determination of total phenolic content using the Folin-C assay: Single-laboratory validation, first action 2017.13. Journal of AOAC international 2018;101(5):1466-1472.
  • 19. Molan AL, Mahdy AS. Iraqi medicinal plants: Total flavonoid contents, free-radical scavenging and bacterial beta-glucuronidase inhibition activities. IOSR Journal of Dental and Medical Sciences 2014;13(5):72-7.
  • 20. Seyrekoğlu F, Temiz H. Effect of Extraction conditions on the phenolic content and DPPH radical scavenging activity of Hypericum perforatum L. TURJFAS 2020;8(1):226-229.
  • 21. Liao KH, et al. Cytotoxicity of graphene oxide and graphene in human erythrocytes and skin fibroblasts. ACS Appl Mater Interfaces 2011;3(7):2607–2615.
  • 22. Cory G. Scratch-Wound Assay. In: Wells C, Parsons M, editors. Cell migration methods in molecular biology, Totowa, NJ: Humana Press; 2011;(769).pp:25-30.
  • 23. Ball JC. A comparison of organic solvent extracts and the fatty acid composition of nonpolar lipids of shed skins from male and female corn snakes (Elaphe guttata guttata). J.Herpetol. 2000: 266-273.
  • 24. Mukherjee S, Dasgupta SC, Gomes A. Effect of Naja naja Laurenti shed skin extract on estrous cycle, hormone-cytokine profiles, histopathology of ovary and uterus of Swiss albino mice. Indian J. Exp. Biol. 2013;51;235-240.
  • 25. Ghosh R, et al. Short-term effect of Naja naja shed skin extract on histopathology of testis, epididymis and vas deferens of swiss albino mice. Uttar pradesh journal of zoology 2022; 43(7):52-59.
  • 26. Park SY, Choi H, Chung JW. Reptiles as promising sources of medicinal natural products for cancer therapeutic drugs. Pharmaceutics 2022; 14(4), 874.
  • 27. Jeyamogan S, et al. Anticancer properties of Asian water monitor lizard (Varanus salvator), python (Malayopython reticulatus) and Tortoise (Cuora kamaroma amboinensis). Anti-Cancer Agents in Medicinal Chemistry-Anti-Cancer Agents 2020; 20(13), 1558-1570.
  • 28. Derakhshani A, et al. Targeting TGF-β-Mediated SMAD Signaling pathway via novel recombinant cytotoxin II: A potent protein from naja naja oxiana venom in Melanoma. Molecules 2020; 25(21), 5148.
  • 29. Thomas Hess C. Checklist for factors affecting wound healing. Adv Skin Wound Care 2011;24(4):192.
  • 30. Menke NB, et al. Impaired wound healing. Clin Dermatol 2007;25(1):19–25.
  • 31. Vakilian S, et al. A Keratin-based biomaterial as a promising dresser for skin wound healing. Wound Med 2019;25(1),100155.
  • 32. Samy RP, et al. Purification of PaTx-II from the Venom of the Australian King Brown Snake and Characterization of Its Antimicrobial and Wound Healing Activities. International Journal of Molecular Sciences 2023; 24(5), 4359.
  • 33. Sen S, et al. Potential broad-spectrum antimicrobial, wound healing, and disinfectant cationic peptide crafted from snake venom. Journal of medicinal chemistry 2023; 66(16), 11555-11572.

Dökülmüş Yılan Derisi (Dolichophis caspius) Sulu Ekstresinin Yara İyileştirici, Sitotoksik ve Antioksidan Özelliklerinin In Vitro Araştırılması

Year 2026, Volume: 16 Issue: 1, 244 - 255, 23.01.2026
https://doi.org/10.31020/mutftd.1749015

Abstract

Amaç: Soğukkanlı sürüngenler olan yılanlar, büyüme sırasında derilerinin kalınlaşması ve deri üzerindeki zararlı parazitlerin uzaklaşması amacıyla mevsimsel olarak ölü deri hücrelerini dökerler. Türkiye'de popüler olarak bilinen bir yılan olan Dolichophis caspius yılın belirli zamanlarında deri dökmektedir. Bu çalışmada, dökülmüş D. caspius derisinin sulu ekstraktlarının toplam fenolik ve toplam flavonoid içerikleri ile antioksidan aktivitesinin belirlenmesi ve bu ekstraktların sitotoksisite ve yara iyileşmesi üzerindeki etkilerinin belirlenmesi amaçlanmıştır.
Yöntem: Dökülmüş yılan derileri distile su kullanılarak ekstrakte edilmiştir. Toplam fenolik içeriği, toplam flavonoid içeriği ve antioksidan aktivitesi sırasıyla Folin-Ciocalteu yöntemi, alüminyum-klorür kolorimetrik deneyi ve DPPH yöntemi ile değerlendirilmiştir. Sitotoksik aktivite MTT testi ile, in vitro yara iyileşmesi üzerindeki etki çizik testi ile belirlenmiştir.
Bulgular: Ekstraktın toplam fenolik içeriği, toplam flavonoid içeriği ve antioksidan aktivitesi sırasıyla 681±46 µg GAE/g, 442.35±13 µg CE/g ve %13.06 olarak hesaplanmıştır. IC50 değerleri BJ dermal fibroblast hücreleri ve SK-MEL-30 melanoma hücreleri için 72. saatte sırasıyla 2972±179 µg/mL ve 2657±128 µg/mL olarak bulunmuştur. Çalışılan dozlarda BJ hücrelerindeki yara kapanma oranları kontrol grubu ile karşılaştırıldığında ekstraktların yara iyileşmesini hızlandırıcı bir etkiye sahip olmadığı görülmektedir.
Sonuç: Dökülmüş yılan derilerinin barındırdıkları potansiyel aktif biyomoleküller sayesinde sitotoksisite ve yara iyileştirme çalışmalarında umut verici sonuçlar doğuracağı tespit edilmiştir.

Project Number

1919B012325139

References

  • 1. Iowa [Internet] Department of Natural Resource. Why do snakes shed their skin? [updated 2023 Nov 02; cited 2025 Jul 23]. Available from: https://www.iowadnr.gov/About-DNR/DNR-News-Releases/ArticleID/158/Why-Do-Snakes-Shed-Their-Skin
  • 2. Sakalli V, et al. Investigation of the usability of snake shirts in the detection of morphological differences between snake species. Eskisehir Technical University Science and Technology Journal-C Life Sciences and Biotechnology 2020;9(1):26-34.
  • 3. Sinmez C, et al. Investigation of immunomodulatory and cytotoxic effects of shed snake skin (Elaphe sauromates) extract. Front.pharmacol. 2024;15,1270970.
  • 4. Andonov K, et al. Analyses of skin secretions of Vipera ammodytes (Linnaeus, 1758) (Reptilia: Serpentes), with focus on the complex compounds and their possible role in the chemical communication. Molecules 2020;25(16):3622.
  • 5. Demı̇rcı̇ ÜÖ. Snake in historical dialects. Electronic Turkish Studies 2014;9(5).
  • 6. Dönmez E. Oral snake skı̇n resultı̇ng in anaphylaxı̇s: How and why? J Emerg Med Case Rep 2020;11(4):95–97.
  • 7. Sakalli V, Çömelekoğlu Ü. Determination of the elemental content of snake shirts used in the treatment of warts in Anatolian folkloric medicine. Mersin University Faculty of Medicine Lokman Hekim Journal of History of Medicine and Folkloric Medicine 2019;9(3):312-316.
  • 8. Khalili F. Studies on the morphology of Dolichophis caspius (Gmeli̇n, 1789) (Serpentes: Colubridae) distributed in western Anatolia, Ege University Biology Department Master Thesis, 2018.
  • 9. Marcov SDC, David A. Dolichophis caspius (Serpentes: Colubridae) in Romania: New distribution records from the northern limit of its range. Turk J Zool 2010:34(1):119-121.
  • 10. Pourmorad F, Hosseinimehr SJ, Shahabimajd N. Antioxidant activity, phenol and flavonoid contents of some selected Iranian medicinal plants. Afr J Biotechnol 2006;5(11).
  • 11. Adam M, Rasad MSBA. Expression of matrix metalloproteinase-13 in human skin melanoma cancer treated by Baccaurea angulata in vitro. J basic appl Res Biomed 2025;1(1):21-28.
  • 12. Yetişgi̇n A. Effects of nanomaterials on stem cell fate determination. Yıldız Technical University Bioengineering Department Graduation Thesis 2018.
  • 13. Guo S, DiPietro LA. Factors affecting wound healing. J Dent Res 2010;89(3):219–229.
  • 14. Gonzalez AC, et al. Wound healing - A literature review. Anais Brasileiros de Dermatologia 2016;91(5):614–620.
  • 15. Öztopalan DF, Recep I, Durmmuş AS. The role of growth factors and cytokines in wound healing. Journal of Dicle University Veterinary Faculty 2017;10(1):83-88.
  • 16. Stamm A, et al. In vitro wound healing assays - state of the art. BioNanoMaterials 2016;17(1-2):79-87.
  • 17. 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):329–333.
  • 18. Kupina S, et al. Determination of total phenolic content using the Folin-C assay: Single-laboratory validation, first action 2017.13. Journal of AOAC international 2018;101(5):1466-1472.
  • 19. Molan AL, Mahdy AS. Iraqi medicinal plants: Total flavonoid contents, free-radical scavenging and bacterial beta-glucuronidase inhibition activities. IOSR Journal of Dental and Medical Sciences 2014;13(5):72-7.
  • 20. Seyrekoğlu F, Temiz H. Effect of Extraction conditions on the phenolic content and DPPH radical scavenging activity of Hypericum perforatum L. TURJFAS 2020;8(1):226-229.
  • 21. Liao KH, et al. Cytotoxicity of graphene oxide and graphene in human erythrocytes and skin fibroblasts. ACS Appl Mater Interfaces 2011;3(7):2607–2615.
  • 22. Cory G. Scratch-Wound Assay. In: Wells C, Parsons M, editors. Cell migration methods in molecular biology, Totowa, NJ: Humana Press; 2011;(769).pp:25-30.
  • 23. Ball JC. A comparison of organic solvent extracts and the fatty acid composition of nonpolar lipids of shed skins from male and female corn snakes (Elaphe guttata guttata). J.Herpetol. 2000: 266-273.
  • 24. Mukherjee S, Dasgupta SC, Gomes A. Effect of Naja naja Laurenti shed skin extract on estrous cycle, hormone-cytokine profiles, histopathology of ovary and uterus of Swiss albino mice. Indian J. Exp. Biol. 2013;51;235-240.
  • 25. Ghosh R, et al. Short-term effect of Naja naja shed skin extract on histopathology of testis, epididymis and vas deferens of swiss albino mice. Uttar pradesh journal of zoology 2022; 43(7):52-59.
  • 26. Park SY, Choi H, Chung JW. Reptiles as promising sources of medicinal natural products for cancer therapeutic drugs. Pharmaceutics 2022; 14(4), 874.
  • 27. Jeyamogan S, et al. Anticancer properties of Asian water monitor lizard (Varanus salvator), python (Malayopython reticulatus) and Tortoise (Cuora kamaroma amboinensis). Anti-Cancer Agents in Medicinal Chemistry-Anti-Cancer Agents 2020; 20(13), 1558-1570.
  • 28. Derakhshani A, et al. Targeting TGF-β-Mediated SMAD Signaling pathway via novel recombinant cytotoxin II: A potent protein from naja naja oxiana venom in Melanoma. Molecules 2020; 25(21), 5148.
  • 29. Thomas Hess C. Checklist for factors affecting wound healing. Adv Skin Wound Care 2011;24(4):192.
  • 30. Menke NB, et al. Impaired wound healing. Clin Dermatol 2007;25(1):19–25.
  • 31. Vakilian S, et al. A Keratin-based biomaterial as a promising dresser for skin wound healing. Wound Med 2019;25(1),100155.
  • 32. Samy RP, et al. Purification of PaTx-II from the Venom of the Australian King Brown Snake and Characterization of Its Antimicrobial and Wound Healing Activities. International Journal of Molecular Sciences 2023; 24(5), 4359.
  • 33. Sen S, et al. Potential broad-spectrum antimicrobial, wound healing, and disinfectant cationic peptide crafted from snake venom. Journal of medicinal chemistry 2023; 66(16), 11555-11572.
There are 33 citations in total.

Details

Primary Language English
Subjects Veterinary Sciences (Other)
Journal Section Research Article
Authors

Faruk Başkurt 0009-0005-0434-3910

Ayşegül İnam 0000-0002-9411-1232

Furkan Ozan Çöven 0000-0001-7696-3418

Tülay Öncü Öner 0000-0001-6180-5409

Project Number 1919B012325139
Submission Date July 26, 2025
Acceptance Date October 31, 2025
Publication Date January 23, 2026
Published in Issue Year 2026 Volume: 16 Issue: 1

Cite

APA Başkurt, F., İnam, A., Çöven, F. O., Öncü Öner, T. (2026). In vitro Investigation of Wound Healing, Cytotoxic, Antioxidant Properties of Shed Snake Skin (Dolichophis caspius) Aqueous Extract. Mersin Üniversitesi Tıp Fakültesi Lokman Hekim Tıp Tarihi Ve Folklorik Tıp Dergisi, 16(1), 244-255. https://doi.org/10.31020/mutftd.1749015
AMA Başkurt F, İnam A, Çöven FO, Öncü Öner T. In vitro Investigation of Wound Healing, Cytotoxic, Antioxidant Properties of Shed Snake Skin (Dolichophis caspius) Aqueous Extract. Mersin Üniversitesi Tıp Fakültesi Lokman Hekim Tıp Tarihi ve Folklorik Tıp Dergisi. January 2026;16(1):244-255. doi:10.31020/mutftd.1749015
Chicago Başkurt, Faruk, Ayşegül İnam, Furkan Ozan Çöven, and Tülay Öncü Öner. “In Vitro Investigation of Wound Healing, Cytotoxic, Antioxidant Properties of Shed Snake Skin (Dolichophis Caspius) Aqueous Extract”. Mersin Üniversitesi Tıp Fakültesi Lokman Hekim Tıp Tarihi Ve Folklorik Tıp Dergisi 16, no. 1 (January 2026): 244-55. https://doi.org/10.31020/mutftd.1749015.
EndNote Başkurt F, İnam A, Çöven FO, Öncü Öner T (January 1, 2026) In vitro Investigation of Wound Healing, Cytotoxic, Antioxidant Properties of Shed Snake Skin (Dolichophis caspius) Aqueous Extract. Mersin Üniversitesi Tıp Fakültesi Lokman Hekim Tıp Tarihi ve Folklorik Tıp Dergisi 16 1 244–255.
IEEE F. Başkurt, A. İnam, F. O. Çöven, and T. Öncü Öner, “In vitro Investigation of Wound Healing, Cytotoxic, Antioxidant Properties of Shed Snake Skin (Dolichophis caspius) Aqueous Extract”, Mersin Üniversitesi Tıp Fakültesi Lokman Hekim Tıp Tarihi ve Folklorik Tıp Dergisi, vol. 16, no. 1, pp. 244–255, 2026, doi: 10.31020/mutftd.1749015.
ISNAD Başkurt, Faruk et al. “In Vitro Investigation of Wound Healing, Cytotoxic, Antioxidant Properties of Shed Snake Skin (Dolichophis Caspius) Aqueous Extract”. Mersin Üniversitesi Tıp Fakültesi Lokman Hekim Tıp Tarihi ve Folklorik Tıp Dergisi 16/1 (January2026), 244-255. https://doi.org/10.31020/mutftd.1749015.
JAMA Başkurt F, İnam A, Çöven FO, Öncü Öner T. In vitro Investigation of Wound Healing, Cytotoxic, Antioxidant Properties of Shed Snake Skin (Dolichophis caspius) Aqueous Extract. Mersin Üniversitesi Tıp Fakültesi Lokman Hekim Tıp Tarihi ve Folklorik Tıp Dergisi. 2026;16:244–255.
MLA Başkurt, Faruk et al. “In Vitro Investigation of Wound Healing, Cytotoxic, Antioxidant Properties of Shed Snake Skin (Dolichophis Caspius) Aqueous Extract”. Mersin Üniversitesi Tıp Fakültesi Lokman Hekim Tıp Tarihi Ve Folklorik Tıp Dergisi, vol. 16, no. 1, 2026, pp. 244-55, doi:10.31020/mutftd.1749015.
Vancouver Başkurt F, İnam A, Çöven FO, Öncü Öner T. In vitro Investigation of Wound Healing, Cytotoxic, Antioxidant Properties of Shed Snake Skin (Dolichophis caspius) Aqueous Extract. Mersin Üniversitesi Tıp Fakültesi Lokman Hekim Tıp Tarihi ve Folklorik Tıp Dergisi. 2026;16(1):244-55.

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