International Journal of Biological Macromolecules 241 (2023) 124529 Available online 20 April 2023 0141-8130/© 2023 Elsevier B.V. All rights reserved. Design, optimization and characterization of a novel antibacterial chitosan-based hydrogel dressing for promoting blood coagulation and full-thickness wound healing: A biochemical and biophysical study Samaneh Sadat Mirhaji a, 1 , Marjan Soleimanpour a, 1 , Hossein Derakhshankhah b , Samira Jafari b , Fatemeh Mamashli a , Meisam Rooki c , Mohammad Reza Karimi d , Hadi Nedaei a , Mitra Pirhaghi a , Hamidreza Motasadizadeh e , Atiyeh Ghasemi a , Maryam Sadat Nezamtaheri a , Fatemeh Saadatpour f , Bahram Goliaei a , C´ edric Delattre g, h , Ali Akbar Saboury a, * a Institute of Biochemistry and Biophysics, University of Tehran, Tehran, Iran b Pharmaceutical Sciences Research Center, Health Institute, Kermanshah University of Medical Sciences, Kermanshah, Iran c Department of Life Science Engineering, Faculty of New Science and Technologies, University of Tehran, Iran d School of Chemistry, College of Science, University of Tehran, Tehran, Iran e Department of Pharmaceutical Nanotechnology, Faculty of Pharmacy, Tehran University of Medical Sciences, Tehran, Iran f School of Biology, College of Science, University of Tehran, Tehran, Iran g Institut Universitaire de France (IUF), 1 rue Descartes, 75005 Paris, France h Universit´ e Clermont Auvergne, CNRS, Clermont Auvergne INP, Institut Pascal, F-63000 Clermont-Ferrand, France A R T I C L E INFO Keywords: Chitosan Hydrogel Wound healing ABSTRACT The use of hydrogel dressings has become increasingly popular as a scaffold for skin tissue engineering. Herein, we have developed an innovative wound dressing using chitosan, fbrinogen, nisin, and EDTA as an effective antibacterial scaffold for wound treatment. The structural and functional characteristics of the hydrogel, including morphology, mechanical strength, drug encapsulation and release, swelling behaviors, blood coagu- lation, cytotoxicity, and antibacterial activity, were studied. Spectroscopic studies indicated that the attachment of chitosan to fbrinogen is associated with minimal change in its secondary structure; subsequently, at higher temperatures, it is expected to preserve fbrinogen's conformational stability. Mechanical and blood coagulation analyses indicated that the incorporation of fbrinogen into the hydrogel resulted in accelerated clotting and enhanced mechanical properties. Our cell studies showed biocompatibility and non-toxicity of the hydrogel along with the promotion of cell migration. In addition, the prepared hydrogel indicated an antibacterial behavior against both Gram-positive and Gram-negative bacteria. Interestingly, the in vivo data revealed enhanced tissue regeneration and recovery within 17 days in the studied animals. Taken together, the results obtained from in vitro and histological assessments indicate that this innovatively designed hydrogel shows good potential as a candidate for wound healing. 1. Introduction When the epithelial continuity of the skin is disrupted, either phys- ically or thermally, it is considered a wound. This provides an envi- ronment for bacteria to grow, potentially resulting in multiple infections in the wound area [1,2]. There are various types of wounds, ranging from superfcial scratches to acute injuries resulting from an accident or surgery. The healing time of a wound depends on diverse parameters, including the size and depth of the wound, the level of blood fow and oxygenation at the wound site, and the presence of bacteria in the area of the wound. In the case of chronic wounds, it can take approximately eight to twelve weeks to heal [3]. Being dynamic and complex, wound healing consists of four different steps: (i) coagulation and homeostasis occurring * Corresponding author. E-mail address: saboury@ut.ac.ir (A.A. Saboury). 1 These authors contributed equally to this work. Contents lists available at ScienceDirect International Journal of Biological Macromolecules journal homepage: www.elsevier.com/locate/ijbiomac https://doi.org/10.1016/j.ijbiomac.2023.124529 Received 13 January 2023; Received in revised form 14 April 2023; Accepted 16 April 2023