Formulation and Evaluation of Clove Extract-Loaded Chitosan Hydrogel on the Mouse Fibroblast Cell Line

Subject Areas : Journal of Animal Biology

Sevda Shemshad ¹ , Fereshteh Sharifi ²

1 - Department of Biology, Central Tehran Branch, Islamic Azad University, Tehran, Iran
2 - Department of Biology, Central Tehran Branch, Islamic Azad University, Tehran, Iran

Received: 2024-09-24 Accepted : 2026-01-28 Published : 2025-06-18

Keywords: Clove extract, Hydrogel, Chitosan, Wound dressing, Antibacterial,

Abstract :

Severe skin damage or defects can cause various problems in living animates, which could affect a patient's life. Developing wound dressings with acceptable antibacterial activity, besides biocompatibility and biodegradability, is essential in developing wound healing applications for decreasing or limiting synthetic antibiotic abuse. The present study aimed to design, prepare, and develop a functional hydrogel structure based on chitosan (CS) containing clove extract (CEO) with enhanced antibacterial activity, non-toxicity, and wound-healing potential. The hydrogel structure was studied using a scanning electron microscope (SEM) and Fourier transform infrared (FTIR). In addition, the cytotoxicity of the designed hydrogel was assayed by the MTT test. Based on the SEM, adding clove extract increased the pore size of the CS hydrogel. FTIR analysis confirmed the presence and mixture of clove extract with CS in the hydrogel structure. Our investigations displayed that the clove extract-loaded CS hydrogel did not have cytotoxic effects on the mouse fibroblast (L929) cells. In addition, the designed hydrogel showed antibacterial activity against Staphylococcus aureus and Escherichia coli. According to the results, CS/CEO hydrogel is a great potential candidate for antibiotic-free wound healing applications.

 

 

References:

1. Anjum S, Arora A, Alam MS, Gupta B. Development of antimicrobial and scar preventive chitosan hydrogel wound dressings. Int J Pharm. 2016;508(1-2):92-101.
2. Ma L, Gao C, Mao Z, Zhou J, Shen J, Hu X, Han C. Collagen/chitosan porous scaffolds with improved biostability for skin tissue engineering. Biomaterials. 2003; 24(26):4833-4841.
3. Akbari A, Rabbani S, Irani S, Zandi M, Sharifi F, Ameli F, Mohamadali M. In vitro and in vivo study of carboxymethyl chitosan/polyvinyl alcohol for wound dressing application. J Appl Polym Sci. 2022; 139(10):51764.
4. Khorasani MT, Joorabloo A, Adeli H, Milan PB, Amoupour M. Enhanced antimicrobial and full-thickness wound healing efficiency of hydrogels loaded with heparinized ZnO nanoparticles: In vitro and in vivo evaluation. Int J Biol Macromol. 2021;166:200-212..
5. Adeli H, Khorasani MT, Parvazinia M. Wound dressing based on electrospun PVA/chitosan/starch nanofibrous mats: Fabrication, antibacterial and cytocompatibility evaluation and in vitro healing assay. Int J Biol Macromol. 2019; 122:238-254.
6. Kabirkoohian A, Bakhshi H, Irani S, Sharifi F. Chemical Immobilization of Carboxymethyl Chitosan on Polycaprolactone Nanofibers as Osteochondral Scaffolds. Appl Biochem Biotechnol. 2023;195(6):3888-3899.
7. El-Samad LM, Hassan MA, Basha AA, El-Ashram S, Radwan EH, Abdul Aziz KK, Tamer TM, Augustyniak M, El Wakil A. Carboxymethyl cellulose/sericin-based hydrogels with intrinsic antibacterial, antioxidant, and anti-inflammatory properties promote re-epithelization of diabetic wounds in rats. Int J Pharm. 2022; 629:122328.
8. Feng W, Wang Z. Biomedical applications of chitosan-graphene oxide nanocomposites. iScience. 2021; 25(1):103629.
9. Mourya VK, Inamdar NN, Tiwari A. Carboxymethyl chitosan and its applications. Advanced Materials Letters, Review Article Adv. Mat. Lett. 2010; 1(1):11-33.
10. Fei Liu X, Lin Guan Y, Yang DZ, Li, Z. Yao KD. Antibacterial action of chitosan and carboxymethylated chitosan. J Appl Polym Sci. 2001;79(7):1324-1335.
11. Li B, Cui J, Xu T, Xu Y, Long M, Li J, Liu M, Yang T, Du Y, Xu Q. Advances in the preparation, characterization, and biological functions of chitosan oligosaccharide derivatives: A review. Carbohydr Polym. 2024;332:121914.
12. Dreiss CA. Hydrogel design strategies for drug delivery. Currt Opin Colloid Interface Sci. 2020;48:1-17.
13. Zhou Y, Yang H, Liu X, Mao J, Gu S, Xu W. Electrospinning of carboxyethyl chitosan/poly(vinyl alcohol)/silk fibroin nanoparticles for wound dressings. Int J Biol Macromol. 2013;53:88-92..
14. Tamer TM, Alsehli MH, Omer AM, Afifi TH, Sabet MM, Mohy-Eldin MS, Hassan MA. Development of Polyvinyl Alcohol/Kaolin Sponges Stimulated by Marjoram as Hemostatic, Antibacterial, and Antioxidant Dressings for Wound Healing Promotion. Int J Mol Sci. 2021;22(23): 13050.
15. Moeini A, Pedram P, Makvandi P, Malinconico M, Gomez d'Ayala G. Wound healing and antimicrobial effect of active secondary metabolites in chitosan-based wound dressings: A review. Carbohydr Polym. 2020; 233:115839..
16. Sharifi F, Hasani M, Atyabi SM, Yu B, Ghalandari B, Li D, et al. Mesenchymal stem cells encapsulation in chitosan and carboxymethyl chitosan hydrogels to enhance osteo-differentiation. Mol Biol Rep. 2022; 49(12):12063-12075..
17. Zhang M, Yang M, Woo MW, Li Y, Han W, Dang X. High-mechanical strength carboxymethyl chitosan-based hydrogel film for antibacterial wound dressing. Carbohydr Polym. 2021;256:117590.
18. Caldwell MD. Bacteria and Antibiotics in Wound Healing. Surg Clin North Am. 2020; 100(4):757-776.
19. Zhang J, Ge J, Xu Y, Chen J, Zhou A, Sun L, Gao Y, Zhang Y, Gu T, Ning X. Bioactive multi-engineered hydrogel offers simultaneous promise against antibiotic resistance and wound damage. Int J Biol Macromol. 2020; 164:4466-4474..
20. Banerjee K, Madhyastha H, Sandur VR, Manikandanath NT, Thiagarajan N, Padma T. Anti-inflammatory and wound healing potential of a clove oil emulsion. Colloids Surf B Biointerfaces. 2020; 193:111102.
21. Ali R, Khamis T, Enan G, El-Didamony G, Sitohy B, Abdel-Fattah G. The healing capability of clove flower extract (CFE) in streptozotocin-induced (STZ-induced) diabetic rat wounds infected with multidrug resistant bacteria. Molecules. 2022;27(7): 2270.
22. Mohammadnezhad J, Khodabakhshi-Soreshjani F, Bakhshi H. Preparation and evaluation of chitosan-coated eggshell particles as copper (II) biosorbent. Desalin. Water Treat. 2016; 57(4):1693-1704.
23. Ray M, Pal K, Anis A, Banthia AK. Development and characterization of chitosan based polymeric hydrogel membranes. Des. Monomers Polym. 2010; 13(3):193-206.
24. Sharifi F, Atyabi SM, Norouzian D, Zandi M, Irani S, Bakhshi H. Polycaprolactone/carboxymethyl chitosan nanofibrous scaffolds for bone tissue engineering application. Int J Biol Macromol. 2018;115:243-248.
25. González-Rivera J, Duce C., Falconieri D, Ferrari C, Ghezzi L, Piras A, et al. Coaxial microwave assisted hydrodistillation of essential oils from five different herbs (lavender, rosemary, sage, fennel seeds and clove buds): Chemical composition and thermal analysis. Innovative Innov Food Sci Emerg Technol, 2016;33:308-318.
26. Parham S, Zargar Kharazi A. Cellulosic textile/clove nanocomposite as an antimicrobial wound dressing: In vitro and in vivo study. Colloids Surf B Biointerfaces. 2022;217:112659.
27. Liang H, Mirinejad MS, Asefnejad A, Baharifar H, Li X, Saber-Samandari S, et al. Fabrication of tragacanthin gum-carboxymethyl chitosan bio-nanocomposite wound dressing with silver-titanium nanoparticles using freeze-drying method. Mater Chem Phys. 2022;279(10):125770
28. Guo W, Ding X, Zhang H, Liu Z, Han Y, Wei Q, Okoro OV, Shavandi A, Nie L. Recent Advances of Chitosan-Based Hydrogels for Skin-Wound Dressings. Gels. 2024;10(3):175.
29. Sivanesan I, Hasan N, Muthu M, Blessing G, Gopal J, Chun S, Shin J, Oh JW. Exploring the impact of chitosan composites as artificial organs. Polymers. 2022;14(8):1587.
30. Shah A, Ashames AA, Buabeid M A, Murtaza G. Synthesis, in vitro characterization and antibacterial efficacy of moxifloxacin-loaded chitosan-pullulan-silver-nanocomposite films. J Drug Deliv Sci Technol. 2020;55:101366.
31. Omidi M, Yadegari A, Tayebi L. Wound dressing application of pH-sensitive carbon dots/chitosan hydrogel. RSC Adv. 2017;7(18):10638-49.
32. Arafa AA, Nada AA, Ibrahim AY, Sajkiewicz P, Zahran MK, Hakeim OA. Preparation and characterization of smart therapeutic pH-sensitive wound dressing from red cabbage extract and chitosan hydrogel. Int J Biol Macromol. 2021;182: 1820-1831.
33. Unalan I, Endlein SJ, Slavik B, Buettner A, Goldmann WH, Detsch R, Boccaccini AR. Evaluation of Electrospun Poly(ε-Caprolactone)/Gelatin Nanofiber Mats Containing Clove Essential Oil for Antibacterial Wound Dressing. Pharmaceutics. 2019;11(11):570.
34. Hameed M, Rasul A, Waqas MK, Saadullah M, Aslam N, Abbas G, Latif S, Afzal H, Inam S, Akhtar Shah P. Formulation and evaluation of a clove oil-encapsulated nanofiber formulation for effective wound-healing. Molecules, 2021; 26(9):2491.