Hemostatic Properties of Green Hydrogel Synthesized from Chitosan/Gelatin Polymers and Hydroalcoholic Extract of Juglans Regia L: An In-Silico and In-Vitro Study
Abstract
Objectives: Quercetin-3-O-rhamnoside, as an effective antioxidant, exists in walnut leaves and is supposed to maintain the structural integrity of the Band 3 protein of Red blood cell (RBCs). In this context, an attempt was made to explore in-silico and in-vitro hemostatic effects of chitosan/gelatin hydrogels containing hydro-alcoholic extract of Iranian walnut leaves (Juglans regia L.), rich in quercetin-3-O-rhamnoside.
Methods: In-silico molecular docking predicted the potential interaction between quercetin-3-O-rhamnoside and band 3 protein. Hydroalcoholic extraction (1:1 water: 70%ethanol) of walnut leaves was performed in a Soxhlet extractor. The hemostatic activity from various extract concentrations (0%, 2.5%, 5%, 10%, and 20% v/v) was assayed using prothrombin time test on citrated blood samples. Gelatin/Chitosan hydrogels (3:1 and 2:1) loaded with the hemostatic concentrations were prepared by the casting method, and their adhesion ability to RBCs was tested by measuring the concentration of free hemoglobin in comparison with controls such as sterile gauze and free-hydrogel positive controls.
Results: Quercetin-3-O-rhamnoside exhibited a high affinity for Band 3 protein, presenting a binding energy of -8.39 kcal/mol compared to the standard ligand 4KU with -7.11 kcal/mol. The nine amino acids found to be important in this interaction include THR728, SER465, VAL729, ARG730, ILE531, LYS851, ILE528, PHE532, and PHE792. All hydrogels incorporating with 2.5% v/v of extract could significantly enhance RBC adhesion controls (p < 0.05).
Conclusion: The hydroalcoholic extract from Juglans regia L. may be of value as an antioxidant and hemostatic agent. Quercetin-3-O-rhamnoside is suggested to be extracted from walnut leaves, and directly incorporated into the hydrogel, and its hemostatic and antioxidant properties can be investigated at both in vitro and in vivo levels.
2. Wang J, Zhuang S. Chitosan-based materials: Preparation, modification and application. Journal of Cleaner Production. 2022;355:131825.
3. Harun-Ur-Rashid M, Jahan I, Foyez T, Imran AB. Bio-inspired nanomaterials for micro/nanodevices: a new era in biomedical applications. Micromachines. 2023;14(9):1786.
4. Gavan A, Colobatiu L, Hanganu D, Bogdan C, Olah NK, Achim M, et al. Development and evaluation of hydrogel wound dressings loaded with herbal extracts. Processes. 2022;10(2):242.
5. Bártolo I, Reis RL, Marques AP, Cerqueira MT. Keratinocyte growth factor-based strategies for wound re-epithelialization. Tissue Engineering Part B: Reviews. 2022;28(3):665-76.
6. Kaith BS, Singh A, Sharma AK, Sud D. Hydrogels: synthesis, classification, properties and potential applications—a brief review. Journal of Polymers and the Environment. 2021;29(12):3827-41.
7. Guillén-Carvajal K, Valdez-Salas B, Beltrán-Partida E, Salomón-Carlos J, Cheng N. Chitosan, gelatin, and collagen hydrogels for bone regeneration. Polymers. 2023;15(13):2762.
8. Irfan NI, Zubir AZM, Suwandi A, Haris MS, Jaswir I, Lestari W. Gelatin-based hemostatic agents for medical and dental application at a glance: A narrative literature review. The Saudi Dental Journal. 2022;34(8):699-707.
9. Zhong Y, Hu H, Min N, Wei Y, Li X, Li X. Application and outlook of topical hemostatic materials: a narrative review. Annals of Translational Medicine. 2021;9(7).
10. Wang H, Ding F, Ma L, Zhang Y. Recent advances in gelatine and chitosan complex material for practical food preservation application. International Journal of Food Science & Technology. 2021;56(12):6279-300.
11. Yadav H, Malviya R, Kaushik N. Chitosan in biomedicine: A comprehensive review of recent developments. Carbohydrate Polymer Technologies and Applications. 2024:100551.
12. Xia Y, Yang R, Wang H, Li Y, Fu C. Application of chitosan-based materials in surgical or postoperative hemostasis. Frontiers in Materials. 2022;9:994265.
13. Kuhn V, Diederich L, Keller Iv TCS, Kramer CM, Lückstädt W, Panknin C, et al. Red blood cell function and dysfunction: redox regulation, nitric oxide metabolism, anemia. Antioxidants & redox signaling. 2017;26(13):718-42.
14. Kim D-C. Identification of quercetin as a potential band 3 protein antioxidant using ektacytometry and in silico molecular docking technology. Intl J Appl Eng Res. 2017;12:8812-6.
15. Zhao M-H, Jiang Z-T, Liu T, Li R. Flavonoids in Juglans regia L. leaves and evaluation of in vitro antioxidant activity via intracellular and chemical methods. The Scientific World Journal. 2014;2014(1):303878.
16. Das SK, Parandhaman T, Dey MD. Biomolecule-assisted synthesis of biomimetic nanocomposite hydrogel for hemostatic and wound healing applications. Green Chemistry. 2021;23(2):629-69.
17. Ryall C, Duarah S, Chen S, Yu H, Wen J. Advancements in skin delivery of natural bioactive products for wound management: a brief review of two decades. Pharmaceutics. 2022;14(5):1072.
18. Ghimire S, Sarkar P, Rigby K, Maan A, Mukherjee S, Crawford KE, et al. Polymeric materials for hemostatic wound healing. Pharmaceutics. 2021;13(12):2127.
19. Zhang B, Wang M, Tian H, Cai H, Wu S, Jiao S, et al. Functional hemostatic hydrogels: design based on procoagulant principles. Journal of Materials Chemistry B. 2024.
20. Croitoru A, Ficai D, Craciun L, Ficai A, Andronescu E. Evaluation and exploitation of bioactive compounds of walnut, Juglans regia. Current Pharmaceutical Design. 2019;25(2):119-31.
21. Taheri P, Jahanmardi R, Koosha M, Abdi S. Physical, mechanical and wound healing properties of chitosan/gelatin blend films containing tannic acid and/or bacterial nanocellulose. International journal of biological macromolecules. 2020;154:421-32.
22. de Oliveira Neto D, Barbosa C, Delmondes P. Flavonoid interaction with chitosan: Planning active packing with antioxidant and antimicrobial activity. 2017.
23. Habibpour S, Mokhtari M, Sharifi E. Effect of hydroalcoholic walnut (Juglans regia) leaf extract on hematological parameters in rat with hypothyroidism. 2014.
24. Nasiry D, Khalatbary AR, Ghaemi A, Ebrahimzadeh MA, Hosseinzadeh MH. Topical administration of Juglans regia L. leaf extract accelerates diabetic wound healing. BMC Complementary Medicine and Therapies. 2022;22(1):255.
25. Khan MA, Mujahid M. A review on recent advances in chitosan based composite for hemostatic dressings. International journal of biological macromolecules. 2019;124:138-47.
26. Lan G, Lu B, Wang T, Wang L, Chen J, Yu K, et al. Chitosan/gelatin composite sponge is an absorbable surgical hemostatic agent. Colloids and surfaces B: Biointerfaces. 2015;136:1026-34.
27. ElShiha HY, Tawfik HA-M, AbouSamrah NK, Marzouk H. Efficacy of chitosan and absorbable gelatin sponge on hemostasis and wound healing following tooth extraction “A Comparative Study”. Egypt Dent J. 2012;58:1-5.
28. Sabino RM, Popat KC. Evaluating whole blood clotting in vitro on biomaterial surfaces. Bio-protocol. 2020;10(3):e3505-e.
| Files | ||
| Issue | Vol 2 No 2 (2024) | |
| Section | Original Articles | |
| DOI | https://doi.org/10.18502/abi.v2i2.17939 | |
| Keywords | ||
| Molecular docking quercetin-3-O-rhamnoside red blood cells chitosan gelatin Juglans regia L | ||
| Rights and permissions | |
|
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License. |
