Download full text in PDF

Aminov RF, Omelyanchik LO, Tavrog ML, Aminova AS, Pototska OI. Morphometric changes of hair follicles in rat excisional wounds treated by Hirudo verbana extract. Bìol Tvarin. 2025; 27 (4): 47–52. DOI: 10.15407/animbiol27.04.056.
https://doi.org/10.15407/animbiol27.04.056
Received 15.11.2025 ▪ Revision 15.12.2025 ▪ Accepted 22.12.2025 ▪ Published online 16.02.2026

Morphometric changes of hair follicles in rat excisional wounds treated by Hirudo verbana extract

R. F. Aminov¹, L. O. Omelyanchik¹, M. L. Tavrog², A. S. Aminova¹, O. I. Pototska²
This email address is being protected from spambots. You need JavaScript enabled to view it.

¹Zaporizhzhia National University, 66 University str, Zaporizhzhia, 69011, Ukraine
²Zaporizhzhia State Medical and Pharmaceutical University, 26 Marii Prymachenko Blvd., Zaporizhzhia, 69035, Ukraine

The study investigated the regenerative potential of a water-salt extract of the medicinal leech Hirudo verbana in excisional skin wounds of rats, focusing on its influence on hair follicle restoration. Sixty male rats were divided into control and experimental groups, the latter receiving topical applications of the extract during wound healing. Histological and morphometric analyses were performed on days 3, 7, and 30 after injury. Treated animals showed earlier transition to the proliferative phase, improved skin structure, and increased density and diameter of hair follicles compared with the control group. By day 30, the number of follicles exceeded that of intact skin, indicating activation of follicular neogenesis and maturation. The therapeutic effect is associated with the bioactive compounds of the leech extract, which exhibit anti-inflammatory, anticoagulant, and regenerative activities, improving local circulation and tissue oxygenation. These findings demonstrate that Hirudo verbana extract effectively promotes hair follicle regeneration and accelerates the overall healing of excisional wounds. Due to its natural origin, safety, and multifunctional biological properties, it may serve as a promising biotechnological agent for skin repair and wound management.

Key words: Hirudo verbana, water-salt extract, excisional wound, hair follicle, wound healing, leech therapy, tissue regeneration

1. Alaama M, Kucuk O, Bilir B, Merzouk A, Ghawi AM, Yerer MB, Ahmado MA, Abdualkader AM, Helaluddin ABM. Development of leech extract as a therapeutic agent: A chronological review. Pharmacol Res Modern Chin Med. 2024; 10: 100355. DOI: 1016/j.prmcm.2023.100355.
2. Aminov RF, Aminova AS. Excisional wound morphological characteristics under the influence of medicinal leech biologically active substances. Innov Biosys Bioeng. 2025; 9 (3): 3–13. DOI: 20535/ibb.2025.9.3.322167.
3. Aminov RF. The influence of the water-salt extract of the medicinal leech Hirudo verbana Carena, 1820 on the general course of embryogenesis in rats after intraperitoneal administration. Stud Biol. 2023; 17 (2): 85–94. DOI: 30970/sbi.1702.713.
4. Aminov RF, Aminova AS, Makyeyeva LV, Frolov OK, Fedotov ER, Piatyhorets TV. The effect of water-salt extract of the medicinal leech Hirudo verbana on the regenerative properties of excisional wounds of rat skin. Bìol Tvarin. 2025; 27 (1): 21–26. DOI: 15407/animbiol27.01.021.
5. Ansell DM, Kloepper JE, Thomason HA, Paus R, Hardman MJ. Exploring the “hair growth-wound healing connection”: Anagen phase promotes wound re-epithelialization. J Invest Dermatol. 2011; 131 (2): 518–528. DOI: 1038/jid.2010.291.
6. Baranzini N, Pulze L, Tettamanti G, Acquati F, Grimaldi A. HvRNASET2 regulate connective tissue and collagen I remodeling during wound healing process. Front Physiol. 2021; 12: 632506. DOI: 3389/fphys.2021.632506.
7. De Moura Estevão LRM, Cassini-Vieira P, Leite AGB, de Carvalho Bulhões AAV, da Silva Barcelos L, Evêncio-Neto J. Morphological evaluation of wound healing events in the excisional wound healing model in rats. Bio-Protocol. 2019; 9 (13): e3285. DOI: 21769/BioProtoc.3285.
8. Ito M, Cotsarelis G. Is the hair follicle necessary for normal wound healing? J Invest Dermatol. 2008; 128 (5): 1059–1061. DOI: 1038/jid.2008.86.
9. Junren C, Xiaofang X, Huiqiong Z, Gangmin L, Yanpeng Y, Xiaoyu C, Yuqing G, Yanan L, Yue Z, Fu P, Cheng P. Pharmacological activities and mechanisms of hirudin and its derivatives — A review. Front Pharmacol. 2021; 12: 660757. DOI: 3389/fphar.2021.660757.
10. Karasartova D, Arslan-Akveran G, Sensoz S, Mumcuoglu KY, Taylan-Ozkan A. Hirudo verbana microbiota dynamics: A key factor in hirudotherapy-related infections? Microorganisms. 2025; 13 (4): DOI: 10.3390/microorganisms13040918.
11. Kvist S, Manzano-Marín A, de Carle D, Trontelj P, Siddall ME. Draft genome of the European medicinal leech Hirudo medicinalis (Annelida, Clitellata, Hirudiniformes) with emphasis on anticoagulants. Sci Rep. 2020; 10: 9885. DOI: 1038/s41598-020-66749-5.
12. Luo G, Chen L, Chen M, Mao L, Zeng Q, Zou Y, Xue J, Liu P, Wu Q, Yang S, Liu M. Hirudin inhibits the formation of NLRP3 inflammasome in cardiomyocytes via suppressing oxidative stress and activating mitophagy. Heliyon. 2023; 10 (1): e23077. DOI: 1016/j.heliyon.2023.e23077.
13. Lv Q, Li M, Wen Z, Han Q, Wei L, Chen J, Pan Y. Efficacy and potential mechanism of atherosclerosis prevention by the active components of leech based on network pharmacology combined with animal experiments. Heliyon. 2024; 10 (6): e27461. DOI: 1016/j.heliyon.2024.e27461.
14. Makyeyeva L, Frolov O, Aliyeva O. Morphometric changes in rat periwound skin during healing of excisional wounds after exposure to chronic social stress. Innov Biosys Bioeng. 2025; 9 (1): 13–25. DOI: 20535/ibb.2025.9.1.310092.
15. Mangelsdorf S, Vergou T, Sterry W, Lademann J, Patzelt A. Comparative study of hair follicle morphology in eight mammalian species and humans. Skin Res Technol. 2014; 20 (2): 147–154. DOI: 1111/srt.12098.
16. Marin E, Kornilov DA, Bukhdruker SS, Aleksenko VA, Manuvera VA, Zinovev EV, Kovalev KV, Shevtsov MB, Talyzina AA, Bobrovsky PA, Kuzmichev PK, Mishin AV, Gushchin IY, Lazarev VN, Borshchevskiy VI. Structural insights into thrombolytic activity of destabilase from medicinal leech. Sci Rep. 2023; 13 (1): 6641. DOI: 1038/s41598-023-32459-x.
17. McCarty SM, Percival SL. Proteases and delayed wound healing. Adv Wound Care. 2013; 2 (8): 438–447. DOI: 1089/wound.2012.0370.
18. Nikrou B, Naeimi S, Rassouli M, Ghaffari Khaligh S. The healing of wounds infected with Staphylococcus aureus is accelerated by leech saliva ointment. Iran J Vet Surg. 2025; 20 (1): 67–77. DOI: 30500/ivsa.2024.467335.1411.
19. Peng L, Pan X, Yin G. Natural hirudin increases rat flap viability by anti-inflammation via PARs/p38/NF-κB pathway. Biomed Res Int. 2015; 2015: 597264. DOI: 1155/2015/597264.
20. Popazova O, Aminov R, Ryzhenko O, Makyeyeva L, Filianskyi I, Gorchakova N. Therapeutic efficacy of Hirudo verbana leech extract in a burn wound model. Phytotherapy J. 2025; 3: 53–61. DOI: 32782/2522-9680-2025-3-53.
21. Pulze L, Baranzini N, Acquati F, Marcolli G, Grimaldi A. Dynamic relationship among extracellular matrix and body wall cells in Hirudo verbana Cell Tissue Res. 2024; 396 (2): 213–229. DOI: 10.1007/s00441-024-03874-x.
22. Sig AK, Guney M, Uskudar Guclu A, Ozmen E. Medicinal leech therapy — an overall perspective. Integr Med Res. 2017; 6 (4): 337–343. DOI: 1016/j.imr.2017.08.001.
23. Song Y, Wu C, Zhang X, Bian W, Liu N, Yin S, Yang MF, Luo M, Tang J, Yang X. A short peptide potentially promotes the healing of skin wound. Biosci Rep. 2019; 39 (3): BSR20181734. DOI: 1042/BSR20181734.
24. Varisco PA, Péclat V, van Ness K, Bischof-Delaloye A, So A, Busso N. Effect of thrombin inhibition on synovial inflammation in antigen-induced arthritis. Ann Rheum Dis. 2000; 59 (10): 781–787. DOI: 1136/ard.59.10.781.