The corrective effect of chitosan on hemostasis parameters under hypocoagulation conditions caused by the antiplatelet agent clopidogrel

The effect of chitosan in vitro on the blood plasma of healthy rats and rats with acquired bleeding caused by the use of the antiplatelet agent clopidogrel, which irreversibly inhibits P2Y12 platelet receptors, was studied. It has been shown that the degree of procoagulant activity of chitosan does not depend on the incubation time (5 and 30 minutes at 37_°C) when it is added to the blood plasma pool from healthy animals. However, there was a significant decrease (by 27%) in the fibrin-depolymerization activity of blood plasma, which is even more suppressed (by 42%) with an increase in incubation time. When chitosan was added to hypocoagulation plasma obtained after oral administration of the antiplatelet agent clopidogrel to rats (once, orally at a dose of 5 mg/kg) and incubation for 5 minutes, a pronounced activation of blood clotting was established. This was evidenced by increased values of fibrinogen concentration (by 38%), fibrin polymerization (by 51%) and platelet aggregation (by 32%), while significantly reducing fibrin depolymerization activity of the blood (by 47%). With an extension of the incubation time of hypocoagulation plasma with chitosan, more pronounced effects of this drug were observed, which manifested itself in a significant increase in the concentration of fibrinogen, platelet aggregation, the degree of polymerization of fibrin and a decrease in thrombin time, characterizing the general pathway of blood coagulation. So, the hemostatic drug chitosan leads to a blockade of bleeding phenomena caused by clopidogrel used in the clinic, which was studied for the first time in this work.

1. Shao H., Wu X., Xiao Y., Yang Y., Ma J., Zhou Y., Chen W., Qin S., Yang J., Wang R., Li H. Recent research advances on polysaccharide-, peptide-, and protein-based hemostatic materials: A review. Int. J. Biol. Macromol. 2024;261(Pt. 1):129752.

2. Wang L., Hao F., Tian S., Dong H., Nie J., Ma G. Targeting polysaccharides such as chitosan, cellulose, alginate and starch for designing hemostatic dressings. Carbohydr. Polym. 2022;291:119574.

3. Gholap A., Rojekar S., Kapare H., Vishwakarma N., Raikwar S., Garkal A., Mehta T., Jadhav H., Prajapati M., Annapure U. Chitosan scaffolds: Expanding horizons in biomedical applications. Carbohydr. Polym. 2024;323:121394.

4. Kumar A., Vimal A., Kumar A. Why chitosan? From properties to perspective of mucosal drug delivery. Int. J. Biol. Macromol. 2016;91:615–622.

5. Wang W., Meng Q., Li Q., Liu J., Zhou M., Jin Z., Zhao K. Chitosan derivatives and their application in biomedicine. Int. J. Mol. Sci. 2020;21(2):487.

6. Naveed M., Phil L., Sohail M., Hasnat M., Baig M., Ihsan A., Shumzaid M., Kakar M., Khan T., Akabar M., Hussain M., Zhou Qi-Gang. Chitosan oligosaccharide (COS): An overview. Int. J. Biol. Macromol. 2019;129:827–843.

7. Wang C.-H., Cherng J.-H., Liu C.-C., Fang T.-J., Hong Z.-J., Chang S.-J., Fan G.-Y., Hsu S.-D. Procoagulant and antimicrobial effects of chitosan in wound healing. Int. J. Mol. Sci. 2021;22(13):7067.

8. Zhao J., Qiu P., Wang Y., Wang Y., Zhou J., Zhang B., Zhang L., Gou D. Chitosan-based hydrogel wound dressing: From mechanism to applications, a review. Int. J. Biol. Macromol. 2023;244:125250.

9. Madurantakam P., Bowlin G., Moskowitz K.A. Wound- stat™ topical hemostat reverses clotting diatheses in models of acquired and congenital hemophilia. Blood. 2008;112(11):2267.

10. Quade-Lyssy P., Ungerer C., Milanov P., Seifried E., Schüttrumpf J. Oral gene delivery of factor IX muteins formulated as chitosan nanoparticles for the treatment of hemophilia B. Blood. 2012:120(21):2048.

11. Misgav M., Lubetszki A., Brutman-Bazarini T., Martinowitz U., Kenet G. The hemostatic efficacy of chitosan-pads in hemodialysis patients with significant bleeding tendency. J. Vasc. Access. 2017; 18 (3):220–224.

12. Li X., Ji X., Chen K., Ullah M.W., Li B., Cao J., Xiao L., Xiao J., Yang G. Immobilized thrombin on X-ray radiopaque polyvinyl alcohol/chitosan embolic microspheres for precise localization and topical blood coagulation. Bioact. Mater. 2021; 6 (7):2105–2119.

13. Камская В.Е. Хитозан: структура, свойства и использование. Научн. обозр. Биол. науки. 2016;6:36–42.

14. Wang W., Xue C., Mao X. Chitosan: Structural modification, biological activity and application. Int. J. Biol. Macromol. 2020;164:4532–4546.

15. Huang Y., Zhang Y., Feng L., He L, Guo R., Xue W. Synthesis of N-alkylated chitosan and its interactions with blood. Artif. Cells Nanomed. Biotechnol. 2018;46(3):544–550.

16. Zhang W., Zhong D., Liu Q., Zhang Y. Effect of chitosan and carboxymethyl chitosan on fibrinogen structure and blood coagulation. J. Biomater. Sci. Polym. Ed. 2013;24(13):1549–1563.

17. Толстенков С.А., Дрозд Н.Н., Макаров В.А., Банникова Г.Е. Биоспецифичный электрофорез комплексов между низкомолекулярными гепаринами и хитозанами. Матер. Восьмой Межд. конф. М.: Изд-во ВНИРО; 2006:253–256.

18. Guo X., Sun T., Zhong R., Ma L., You C., Tian M., Li H., Wang C. Effects of chitosan oligosaccharides on human blood components. Front. Pharmacol. 2018;9:1412.

19. Шубина Т.А., Оберган Т.Ю., Ляпина Л.А., Григорьева М.Е. Влияние полисахарида хитозана на плазменный гемостаз: исследование in vitro. Тромбоз, гемостаз и реология. 2022; (4):30–34.

20. Ляпина Л.А., Григорьева М.Е., Оберган Т.Ю., Шубина Т.А. Исследование гемостатических свойств препарата на основе хитозана. Биофарм. журнал. 2022;14(1):3–7.

21. Benesch J., Tengvall P. Blood protein adsorption onto chitosan. Biomaterials. 2002;23(12):2561–2568.

22. Denzinger М., Hinkel H., Kurz J., Hierlemann T., Schlensak Ch., Wendel H.P., Krajewski S. Hemostyptic property of chitosan: Opportunities and pitfalls. Biomed. Mater. Eng. 2016;27(4):353–364.

23. Izaguirre-Avila R., De la Peña-Díaz A., Barinagarrementería-Aldatz F., González-Pacheco H., Ramírez-Gutierrez A.E., Ruiz-Sandoval J.L., Quiroz-Martínez A., Cantú-Brito C. Effect of clopidogrel on platelet aggregation and plasma concentration of fibrinogen in subjects with cerebral or coronary atherosclerotic disease. Clin. Appl. Thromb. Hemost. 2002;8(2):169–177.

24. Kuszynski D., Lauver D. Pleiotropic effects of clopidogrel. Purinergic Signal. 2022; 18(3):253–265.

25. Ляпина Л.А., Григорьева М.Е., Шубина Т.А., Оберган Т.Ю. Основы физиологии и биохимии свертывания крови. М.: Изд-во Моск. ун-та; 2023. 159 с.

26. Okamoto Y., Yano R., Miyatake K., Tomohiro I., Shigemasa Y., Minami S. Effects of chitin and chitosan on blood coagulation. Carbohydr. Polym. 2003;53(3):337–342.

27. Klokkevold P.R., Fukayama H., Sung E.C., Bertolami C.N. The effect ofchitosan (poly-N-acetyl glucosamine) on lingual hemostasis in heparinized rabbits. J. Oral Maxillofac. Surg. 1999; 57(1): 49–52.

28. Кузник Б.И. Клеточные и молекулярные механизмы регуляции системы гемостаза в норме и патологии. Чита: Экспресс-изд-во; 2010. 832 с.

29. Григорьева М.Е., Ляпина Л.А. Регуляторные пептиды как противотромботические агенты. М.: Ким Л.А.; 2021. 108 с.

30. Lyapina L., Grigorjeva M., Lyapin G., Obergan T., Shubina T. Aggregation effects of chitosan in the blood. NJD-iScience. 2021;(61):13–16.

31. Бокарев И.Н. Гематология для практического врача. М.: Медицинское информ. агентство; 2018. 344 c.

32. Rodriguez-Merchan E.C. Local fibrin glue and chitosan-based dressings in haemophilia surgery. Blood Coagul. Fibrinolysis. 2012;23(6):473–476.

33. Olabisi O.S., Nathan A.F. Topical coagulant agents. Surg. Clin. North. Am. 2022;102(1):65–83.