Study on the Preparation of Multifunctional Bacterial Cellulose-Based Wound Dressings and Their In Vitro Hemostatic Performance

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To enhance its functionality, a novel BC-based wound dressing (HBP) was engineered by integrating chitosan quaternary ammonium salt (HACC) and hydroxyapatite (HAP) into BC's 3D structure via soaking, stirring, and freeze-drying techniques. This composite dressing demonstrates significant antibacterial activity, evidenced by distinct inhibition zones in antimicrobial assays, and rapid hemostatic performance, with an in vitro clotting time of 300 ± 5 seconds. Characterized using SEM, XRD, FT-IR, and XPS, HBP exhibits superior physicochemical characteristics and biocompatibility, positioning it as a highly viable option for advanced biomedical wound dressings. Physical sciences/Materials science/Biomaterials Biological sciences/Microbiology/Antimicrobials Bacterial cellulose Chitosan quaternary ammonium salt Hydroxyapatite Antibacterial test In vitro hemostasis Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Introduction Uncontrollable bleeding is a primary cause of death following trauma, often leading to rapid mortality and complications such as coagulopathy, hypothermia, and severe infection 1 , 2 . Current commercial hemostatic materials, commonly used in military settings, are composed of polysaccharides, silicoaluminates, biological products, and peptides 3 .However, these materials are often ineffective for moderate to severe bleeding, may cause side effects, and are relatively expensive 4 – 6 . Given these limitations, there is a critical need for rapid and efficient hemostatic materials suitable for civilian use, highlighting the importance of research in this area. Chitosan quaternary ammonium salt (HACC, Scheme 1a ) is prized for its excellent water solubility, flocculation, biocompatibility, hygroscopicity, moisture retention, and antibacterial properties, making it a versatile material in biomaterials research 7 – 9 . For instance, Min et al. 10 HACC/PVA composite coating modified with quaternary ammonium salt via an eco-friendly solution casting method. It achieves 99% inhibition against E. coli and S. aureus, and features transparency, biodegradability, anti-fogging, and antibacterial properties. Similarly. Mahdi Rahimi et al. 11 developed a quaternized chitosan/nanosilver composite membrane with antimicrobial properties to reduce wound infections. Tests using MTT assay and DAPI staining showed good biocompatibility and antibacterial performance. These studies highlight the potential of HACC as an effective antibacterial agent in non-toxic skin dressings. Its cationic nature allows it to form a barrier that adsorbs and kills bacteria, thereby protecting the wound surface and promoting healing. Hydroxyapatite (HAP, Scheme 1b ) is recognized for its biocompatibility, non-toxicity, non-irritating nature, and lack of immunogenicity, making it resistant to causing aging, sensitization, or carcinogenesis 12 . It significantly contributes to enhancing osteoblast proliferation, bone regeneration, and hemostatic processes 13 , 14 . Emerging research indicates that materials based on HAP can engage with blood constituents to accelerate clotting 15 , 16 . For instance, Noh and colleagues engineered a composite of plant polysaccharides and HAP designed for fracture hemostasis, characterized by its high solubility, swift hemostatic action, and adhesive qualities 17 . Similarly, Tham et al. discovered that a composite material (BoneSeal®), which combines HAP with biodegradable polylactic acid, exhibits dual capabilities in bone restoration and hemostasis, effectively minimizing blood loss from fractures 18 . Consequently, mineral-based hemostatic materials utilizing HAP are emerging as a promising solution for external hemostasis and the facilitation of tissue repair. Bacterial cellulose (BC), produced by microorganisms such as Gluconacetobacter xylinus, is a natural nanofibrous material distinguished by its superior water retention, crystallinity, mechanical strength, and intricate three-dimensional network structure compared to plant-derived cellulose 19 , 20 . BC is highly biocompatible, biodegradable, non-toxic, and hypoallergenic, making it suitable for medical applications 21 . Its breathable nature minimizes the risk of wound infections, and it can be further functionalized to exhibit antimicrobial properties 22 – 24 . The thickness, morphology, and functionality of BC can be precisely engineered, allowing its membranes to be customized for specific hemostatic requirements 25 , 26 . Additionally, BC facilitates wound healing by maintaining a moist microenvironment conducive to tissue regeneration, positioning it as an excellent candidate for advanced wound care dressings in modern medicine 27 , 28 . In this study, HACC and HAP were integrated into the 3D network of BC (derived from kombucha-fermented pear juice) using soaking, stirring, and freeze-drying techniques, resulting in the development of HBP, a composite material with enhanced antibacterial and hemostatic properties. The study results show that the HBP wound dressing has excellent antibacterial properties, with clear inhibition zones against Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, and Streptococcus mutans. Additionally, HBP reduces the hemostatic time to 300 ± 5 seconds by effectively concentrating coagulation factors and platelets, which significantly accelerates clotting compared to pure BC. Materials and methods Materials Kombucha was purchased from Shandong Zhi's Gaibao Biotechnology Co, Ltd. Yeast extract was obtained from Beijing Aoboxing Biotechnology Co, Ltd. Glucose, chitosan quaternary ammonium salt (with a degree of substitution > 98%), and hydroxyapatite (with a purity ≥ 97%) were purchased from Shandong Keyuan Biochemical Co, Ltd. Methods Preparation of BC Films In a 250 mL conical flask, 100 mL of pear juice supernatant was mixed with 6% glucose and 1% yeast extract. After high-temperature sterilization, 10 mL of kombucha inoculum was added, and the flask was sealed with sterilized gauze. The mixture was statically cultured at 30°C for 7 days. The resultant bacterial cellulose (BC) film was washed with distilled water, treated with 1 wt% NaOH for 24 hours to remove impurities, washed again, soaked in 0.5 wt% glacial acetic acid for 24 hours, and finally washed until neutral. The film was then freeze-dried to obtain the purified BC. Preparation of HBC Antibacterial Film BC was immersed in a 2.0% (w/v) HACC solution (100 mL) for 3 days, then washed with deionized water and freeze-dried to obtain the antibacterial BC-HACC film (HBC). Preparation of the Multifunctional Antibacterial Wound Dressing (HBP) Dissolve 1.0 g of the material in 100 mL water, add HBC, and stir magnetically for 3 hours to adhere HAP. Rinse with deionized water and vacuum freeze-dry to obtain the multifunctional BC-based wound dressing (BC-HACC-HAP, named HBP). The technical route is shown in Scheme 1 . X-ray Diffraction Analysis (XRD) XRD tests on BC, HBC, and HAP composites were conducted using a Rigaku Smartlab 9kW diffractometer (Japan), with a scanning range of 5°-80°at 10°/min. Fourier-Transform Infrared Spectroscopy Analysis (FT-IR) FT-IR analysis of BC, HBC, and HAP composites was conducted using a Thermo Scientific Nicolet iS20 spectrometer with ATR method, scanning 16 times over 400–4000 cm − 1 . X-ray Photoelectron Spectroscopy (XPS) XPS analysis of BC, HBC, and HBP composites was conducted using a Shimadzu/Kratos AXIS Ultra DLD spectrometer to measure O, C, Cl, Ca, and P content. Scanning Electron Microscopy Analysis (SEM) SEM analysis of BC, HBC, and HAP composites was performed using a Hitachi Regulus 8100 microscope at 10 kV, with EDS elemental analysis on selected areas. Sample size was 5 mm × 1 mm. Performance Testing of the Material's Water Management System Water Content Test: Water content was measured by weighing samples (M1), freeze-drying them, and reweighing (M2). Wwater Content was calculated using the formula: $$\:{W}_{\text{w}\text{a}\text{t}\text{e}\text{r}\:\text{c}\text{o}\text{n}\text{t}\text{e}\text{n}\text{t}}=\frac{{M}_{1}-{M}_{2}}{{M}_{3}}\times\:100\%$$ Rehydration Test: Samples were soaked in distilled water for 24 hours, surface moisture removed, and weighed (M3). Wrehydration Rate was calculated as: $$\:{W}_{\text{r}\text{e}\text{h}\text{y}\text{d}\text{r}\text{a}\text{t}\text{i}\text{o}\text{n}}=\frac{{M}_{3}-{M}_{2}}{{M}_{3}}\times\:100\%$$ Water Vapor Permeability (WVT) Test: A 50 mL beaker with 10 mm distilled water was covered with the sample, sealed with a rubber band, and tested at 38°C, 50% humidity for 24 hours (effective area: 0.001963 m²). A plastic-wrapped beaker served as control. WVT was calculated using the formula: $$\:WVT=\frac{\varDelta\:m-\varDelta\:m{\prime\:}}{A\bullet\:t}$$ In the formula: ∆m and ∆m₀are weight differences for the sample and blank, A is the effective area (m²), and T is the test duration Thermogravimetric Analysis (TG) TGA of BC, HBC, and HAP composites was performed using a HITACHI STA200 analyzer. Samples (20 mg) were heated from 30°C to 800°C at 20°C/min under nitrogen (50 L/min). Mechanical Property Testing Tensile tests on BC, HBC, and HAP composites were performed using an INSTRON 8872 universal testing machine. Specimens in the form of strips (10 mm × 50 mm) were tested at a crosshead speed of 5 mm/min, with three replicates for each material to obtain average values. In Vitro Hemostasis Test 40 mg of BC, HBC, and HBP were each added to 2 mL of anticoagulated pig blood supplemented with 80 µL of 0.2 mol/L CaCl₂ to neutralize the anticoagulant. Blood coagulation time was measured using a silent mixer at 37°C, with a 45° oscillation and a rotation speed of 5 r. p. m. The anticoagulant used was 10% sodium citrate. Hemolysis Rate Test BC, HBC, and HBP were added to 2 mL of 50× diluted red blood cell suspension and incubated at 37°C for 2 hours. After centrifugation (3000 r. p. m, 10 min), 100 µL supernatant absorbance was measured at 540 nm. Positive control: 0.2% TX-100; negative control: no material. Hemolysis rate was calculated as: $$\:\text{H}\text{e}\text{m}\text{o}\text{l}\text{y}\text{s}\text{i}\text{s}\:\text{R}\text{a}\text{t}\text{e}=\frac{{A}_{s}-{A}_{n}}{{A}_{p}-{A}_{n}}$$ As, Ap, and An represent the hemolysis rates of the experimental, positive, and negative control groups, respectively. Cytotoxicity Test RAW264.7 macrophages were seeded in a 96-well plate and cultured until 80–90% confluence. The medium was then replaced with 100 µL of sample extracts at concentrations of 0, 10, 20, 40, and 80 mg/mL. After 24 hours of incubation, the cells were treated with CCK-8 reagent, and absorbance was measured. Each sample was tested in triplicate, with a blank control included. The culture conditions were maintained at 30°C with 5% CO₂. Antibacterial Performance Evaluation E. coli, P. aeruginosa, and S. aureus cultures were adjusted to 10⁶ CFU/mL using sterile PBS. Aliquots of 100 µL of each bacterial suspension were spread onto LB agar plates. Sterilized samples were placed on the agar surface, and the plates were incubated at 37°C for 24 hours. The sizes of the inhibition zones were measured. Each sample was tested in triplicate, and the results were averaged. S. mutans was processed similarly but incubated for 48 hours, with data recorded accordingly. Results and Discussion Surface morphology of HBP Figure 1 A shows that freeze-dried HBP appears more transparent than pure BC, with a white powdery precipitate on the surface 29 , 30 . The transparency is attributed to the bleaching effect of acetic acid during HBC soaking, while the precipitate results from HAP deposition during the composite process. Figure 1 B reveals that HBP maintains a three-dimensional fibrous structure, with plate-like HAP crystals formed through mineralization. These crystals are relatively large, and some HAP further grows into spherical shapes, unevenly filling the cellulose network 31 . Higher-magnification SEM images (Fig. 1 C) show that these spherical structures are composed of plate-like HAP crystals. Eds Analysis Figure 2 a shows the total distribution of elements in the EDS analysis. Figure 2 b-f are the EDS distribution maps of C, O, Cl, P, and Ca elements, respectively. These results confirm the presence of chlorine (Cl, from HACC) and phosphorus (P) and calcium (Ca, from HAP) on the surface of HBP, indicating that both HACC and HAP have been successfully integrated into the HBP network 32 , 33 . Structural analysis of HBP Figure 3 A displays the XRD patterns of the samples. HBC exhibits enhanced crystallinity compared to BC due to dehydration and molecular rearrangement, although no new peaks emerge (curves a and b). In contrast, HBP, which incorporates HAP, displays characteristic peaks of BC (14.6°, 17.7°, 22.7°) and additional crystal plane peaks of HAP (002), (211), (112), (300), (202), (312), (222), (213), and (304), consistent with the standard card (JCPDS No. 09-0432). The XRD results confirm the presence of HAP on the cellulose, corroborating the FTIR data (curve c) 34 . Figure 3 B illustrates the characteristic FTIR peaks of BC: -OH (3340 cm⁻¹), C-H (2850–2920 cm⁻¹), -CH₃ (1640 cm⁻¹), and cellulose (1309 cm⁻¹). HBC shows a slight shift at 2920 cm⁻¹ and enhanced C-H peaks, indicating the incorporation of HACC into the BC structure. The FTIR spectrum of HBP (curve c) reveals C = O peaks at 1611 cm⁻¹ and P-O peaks at 960 cm⁻¹ and 1021 cm⁻¹, confirming the presence of PO₄³⁻. Figure 3 C and 3 D present XPS analysis, revealing the presence of Cl, P, and Ca elements in the material due to the incorporation of HACC and HAP. The binding energies at Cl 2p, P 2p, and Ca 2p further confirm their successful integration into the BC network 35 – 37 . Performance Analysis of HBP Figure 4 A: DTG curves indicate weight loss between 38–130°C due to moisture evaporation. HBC and HBP degrade at 246°C (attributed to HACC), while HAP raises the decomposition temperature to 429°C and increases residual mass, demonstrating superior thermal stability for HBP 38 . Figure 4 B: Tensile tests reveal BC with a strength of 29.43 ± 0.71 MPa and modulus of 18.59 ± 0.82 MPa. Incorporation of HACC and HAP reduces HBC's strength to 17.98 ± 0.766 MPa and modulus to 10.27 ± 1.06 MPa. However, HBP recovers to a strength of 26.19 ± 2.24 MPa and modulus of 17.92 ± 1.64 MPa, with HAP enhancing mechanical properties by tightening fiber bonds 39 . Figure 4 C: In vitro hemostasis tests show that HBP achieves the fastest clotting time (300 ± 5 s) compared to others (582 ± 4 s, 484 ± 8 s, 513 ± 10 s). HBP's high water absorption and 3D structure concentrate coagulation factors, while HAP releases Ca²⁺ to further promote clotting 40 , 41 . Table 1 presents the water management performance of the samples. BC demonstrated excellent water retention and moisture permeability, with a water content of 98.02 ± 0.10%, rehydration rate of 87.35 ± 0.58%, and water vapor transmission rate (WVTR) of 5335.84 ± 2.39 g/m²·24h. These properties are attributed to its high surface area and porosity. HBC and HBP exhibited slightly lower water retention and rehydration rates (98.82 ± 0.03%, 84.58 ± 0.63% for HBC; 96.22 ± 0.02%, 82.16 ± 0.46% for HBP), but maintained good moisture permeability (6106.76 ± 2.88 g/m²·24h for HBC; 5528.36 ± 4.12 g/m²·24h for HBP). Despite the influence of composite materials, HBP still provided an effective sterile healing environment for wound repair due to its high moisture permeability. Table 1 Water Content, Rehydration Rate, and Water Vapor Transmission Rate (WVT) of BC, HBC, and HBP dry weight(g) water conten(%) rehydratio(%) wvt(g/m2*24h) BC 0.88 ± 0.21 98.02 ± 0.10 87.35 ± 0.58 5335.84 ± 2.39 HBC 0.84 ± 0.30 98.82 ± 0.03 84.58 ± 0.63 6106.76 ± 2.88 HBP 0.87 ± 0.43 96.22 ± 0.02 82.16 ± 0.46 5528.36 ± 4.12 Biocompatibility Analysis of HBP Figure 5 illustrates the biocompatibility of the sample membranes. Figure 5 A shows that BC, HBC, and HBP have hemolysis rates of 1.61%, 1.86%, and 4.24%, respectively, all below the 5% threshold for hemocompatibility 42 . This indicates that the incorporation of HACC and HAP does not damage red blood cells, which remain healthy and evenly distributed, making these materials suitable for biomedical applications involving blood contact. Figure 5 B demonstrates that HBP exhibits good biocompatibility, with high cell survival rates for RAW264.7 macrophages across all extract concentrations, even slightly promoting cell growth. Thus, HBP shows excellent hemocompatibility and no cytotoxicity within the 20–80 mg/mL range. Antibacterial Analysis of HBP Figure 6 presents the antibacterial performance of BC, HBC, and HBP against four common pathogens: Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, and Streptococcus mutans. BC exhibited no antibacterial activity, while HBC and HBP demonstrated clear inhibition zones, indicating strong antibacterial effects. HBP showed superior performance due to the slow release of Ca²⁺ and PO₄³⁻ from HAP, which enhances local ion concentration and boosts antibacterial efficacy 43 , 44 . Thus, HBP effectively prevents bacterial infections when used as a hemostatic dressing. Conclusion This study developed an HBP composite dressing with antibacterial and hemostatic properties by incorporating HACC and HAP into BC's 3D network through soaking, stirring, and freeze-drying. The HBP dressing exhibits no cytotoxicity, meets hemolysis standards, and demonstrates improved thermal stability and tensile strength, making it suitable for medical applications. Compared to BC, HBP effectively prevents bacterial infections during hemostasis and achieves a hemostatic time of 300 ± 5 seconds by promoting blood cell aggregation and activating coagulation at the wound site. Overall, HBP shows potential as a novel multifunctional hemostatic dressing for preventing infections and achieving rapid hemostasis. Declarations Competing interests The authors declare no competing interests. Author Contribution Miaoyi Tian: Data curation, Investigation, Visualization, Writing -original draft, Writing-review & editing. Shengbiao Ma: Data curation, Investigation, Visualization, Writing -original draft. Zhenchao Dong: Investigation, Writing-review & editing. Shenao Wang: Investigation. Xinyu Liu: Investigation. Binhui Liu: Investigation. Huaizhi Cui: Investigation. Mingshuai Ma: Investigation. Hang Zheng: Investigation. Xin Liu: Conceptualization, Formal analysis, Funding acquisition, Methodology, Resources. 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Lv, Y. et al. Evaluation of the antibacterial properties and in-vitro cell compatibilities of doped copper oxide/hydroxyapatite composites. Colloids Surf., B . 209 , 112194 (2022). Scheme 1 Scheme 1 is available in the Supplementary Files section. Additional Declarations No competing interests reported. Supplementary Files TOC.docx Scheme1.jpg Scheme 1. Schematic Diagram of the Preparation Process of HBP Wound Dressing Cite Share Download PDF Status: Published Journal Publication published 22 Apr, 2026 Read the published version in Scientific Reports → Version 1 posted Editorial decision: Revision requested 16 May, 2025 Reviews received at journal 16 May, 2025 Reviewers agreed at journal 28 Apr, 2025 Reviews received at journal 09 Apr, 2025 Reviewers agreed at journal 04 Apr, 2025 Reviewers agreed at journal 31 Mar, 2025 Reviewers invited by journal 20 Mar, 2025 Editor assigned by journal 20 Mar, 2025 Editor invited by journal 20 Mar, 2025 Submission checks completed at journal 20 Mar, 2025 First submitted to journal 11 Mar, 2025 You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-6201748","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":432085118,"identity":"8a157cfd-8904-4f3a-a1af-ef8a30563579","order_by":0,"name":"Miaoyi Tian","email":"","orcid":"","institution":"Harbin University of Science and Technology","correspondingAuthor":false,"prefix":"","firstName":"Miaoyi","middleName":"","lastName":"Tian","suffix":""},{"id":432085119,"identity":"b1591740-c2d6-4f32-866f-05b2238939b4","order_by":1,"name":"Shengbiao Ma","email":"","orcid":"","institution":"Harbin University of Science and Technology","correspondingAuthor":false,"prefix":"","firstName":"Shengbiao","middleName":"","lastName":"Ma","suffix":""},{"id":432085120,"identity":"8d6b6fa0-97df-4fd4-b778-acb6759b69bd","order_by":2,"name":"Zhenchao Dong","email":"","orcid":"","institution":"Harbin University of Science and Technology","correspondingAuthor":false,"prefix":"","firstName":"Zhenchao","middleName":"","lastName":"Dong","suffix":""},{"id":432085121,"identity":"b5fd9064-0855-45f3-976f-f2e104aaa96b","order_by":3,"name":"Shenao Wang","email":"","orcid":"","institution":"Harbin University of Science and Technology","correspondingAuthor":false,"prefix":"","firstName":"Shenao","middleName":"","lastName":"Wang","suffix":""},{"id":432085122,"identity":"b2249377-5224-4e20-ac2b-d9217770dcfb","order_by":4,"name":"Xinyu Liu","email":"","orcid":"","institution":"Harbin University of Science and Technology","correspondingAuthor":false,"prefix":"","firstName":"Xinyu","middleName":"","lastName":"Liu","suffix":""},{"id":432085123,"identity":"1b86efe6-bc06-4c28-b4a9-11269ced4c52","order_by":5,"name":"Binhui Liu","email":"","orcid":"","institution":"Harbin University of Science and Technology","correspondingAuthor":false,"prefix":"","firstName":"Binhui","middleName":"","lastName":"Liu","suffix":""},{"id":432085124,"identity":"8d4ac26a-6f80-4aeb-abbc-d51a919a0814","order_by":6,"name":"Huaizhi Cui","email":"","orcid":"","institution":"Harbin University of Science and Technology","correspondingAuthor":false,"prefix":"","firstName":"Huaizhi","middleName":"","lastName":"Cui","suffix":""},{"id":432085125,"identity":"86f3d6fa-1b07-4f54-9095-64ec2474cea4","order_by":7,"name":"Mingshuai Ma","email":"","orcid":"","institution":"Harbin University of Science and Technology","correspondingAuthor":false,"prefix":"","firstName":"Mingshuai","middleName":"","lastName":"Ma","suffix":""},{"id":432085126,"identity":"1355f6b7-d62b-4ef3-88c9-295755229637","order_by":8,"name":"Hang Zheng","email":"","orcid":"","institution":"Harbin University of Science and Technology","correspondingAuthor":false,"prefix":"","firstName":"Hang","middleName":"","lastName":"Zheng","suffix":""},{"id":432085127,"identity":"c61efa9b-c4e7-4123-9248-d73b6fd152fd","order_by":9,"name":"Xin Liu","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAAv0lEQVRIiWNgGAWjYDACZgbGAwkMNnL8zMwHHxCrhQGoJc1Ysp0t2YBoiw4wMBxO3HCex0yAKOUGx3kPHHjYdtjY+DCDGQNDjU00QS2SzXwJBxLb0uXMDjOkPWA4lpbbQEgLPzOPAVCLtTFQy3EDxobDhLWwQbQwJ25uZmyTIEoL1BbnxA3MzGzEaZFsBmpJOJdmLHGYjdkggRi/GJw/Y/jwRxkwKvvPf3zwocaGsBZUkECa8lEwCkbBKBgFuAAAry88yV4iuesAAAAASUVORK5CYII=","orcid":"","institution":"Harbin University of Science and Technology","correspondingAuthor":true,"prefix":"","firstName":"Xin","middleName":"","lastName":"Liu","suffix":""}],"badges":[],"createdAt":"2025-03-11 09:38:16","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-6201748/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-6201748/v1","draftVersion":[],"editorialEvents":[{"content":"https://doi.org/10.1038/s41598-026-48080-7","type":"published","date":"2026-04-22T15:58:56+00:00"}],"editorialNote":"","failedWorkflow":false,"files":[{"id":79071369,"identity":"1c6fb417-73bd-4b82-82e1-eb9b5521ebd4","added_by":"auto","created_at":"2025-03-24 06:09:35","extension":"jpg","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":68220,"visible":true,"origin":"","legend":"\u003cp\u003eMacroscopic image of HBP (A); SEM images of HBP (B, C);\u003c/p\u003e","description":"","filename":"1.jpg","url":"https://assets-eu.researchsquare.com/files/rs-6201748/v1/b843d34fdc65fb041456de1a.jpg"},{"id":79071589,"identity":"ce41f803-992f-4409-819e-f91958ece33a","added_by":"auto","created_at":"2025-03-24 06:17:35","extension":"jpg","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":102837,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eEDS analysis distribution total chart（a）；Carbon element (b)；Oxygen element (c),；Chlorine element (d)；Phosphorus element (e)；Calcium element (f).\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"2.jpg","url":"https://assets-eu.researchsquare.com/files/rs-6201748/v1/70ba3fcaf082e2eb5cdf3e1b.jpg"},{"id":79071594,"identity":"3f1b6d9f-2018-4844-ab9a-922b40ad41ed","added_by":"auto","created_at":"2025-03-24 06:17:35","extension":"jpg","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":51340,"visible":true,"origin":"","legend":"\u003cp\u003eXRD patterns of BC, HBC, and HBP (A); SEM images of BC, HBC, and HBP (B); XPS elemental analysis of BC, HBC, and HBP (C, D). Curves a, b, and c represent BC, HBC, and HBP, respectively.\u003c/p\u003e","description":"","filename":"3.jpg","url":"https://assets-eu.researchsquare.com/files/rs-6201748/v1/f9121a9352a8e1e4c9a3b908.jpg"},{"id":79071596,"identity":"93c23f27-fe1d-4802-b536-ac318a583eac","added_by":"auto","created_at":"2025-03-24 06:17:35","extension":"jpg","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":39430,"visible":true,"origin":"","legend":"\u003cp\u003eTG curves of BC, HBC, and HBP (A); stress-strain curves of BC, HBC, and HBP (B); in vitro hemostasis time of BC, HBC, and HBP (C). Curves a, b, and c represent BC, HBC, and HBP, respectively.\u003c/p\u003e","description":"","filename":"4.jpg","url":"https://assets-eu.researchsquare.com/files/rs-6201748/v1/121285517d92ad4a17f6722c.jpg"},{"id":79071371,"identity":"e7716070-a918-4fad-a582-7bde388fe542","added_by":"auto","created_at":"2025-03-24 06:09:35","extension":"jpg","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":38939,"visible":true,"origin":"","legend":"\u003cp\u003eHemolysis rate test of BC, HBC, and HBP (A); Cytotoxicity test of BC, HBC, and HBP (B).\u003c/p\u003e","description":"","filename":"5.jpg","url":"https://assets-eu.researchsquare.com/files/rs-6201748/v1/f9ec7eaaed183a44c4ef9893.jpg"},{"id":79071381,"identity":"e424a81b-ae7c-4715-aef4-b359a2a81418","added_by":"auto","created_at":"2025-03-24 06:09:35","extension":"jpg","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":38098,"visible":true,"origin":"","legend":"\u003cp\u003eAntibacterial test of BC, HBC, and HBP. Samples 1, 2, and 3 are BC, HBC, and HBP. A, B, C, and D correspond to Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa, and Streptococcus mutans.\u003c/p\u003e","description":"","filename":"6.jpg","url":"https://assets-eu.researchsquare.com/files/rs-6201748/v1/7b44ab001b97dfbe1d04cc35.jpg"},{"id":107928163,"identity":"45b3a943-8cff-451a-bbf7-4ac296857256","added_by":"auto","created_at":"2026-04-27 16:08:42","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":630221,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-6201748/v1/c777266e-77ae-4032-8922-54069d07e923.pdf"},{"id":79071393,"identity":"3aaf656a-0ba2-483d-89d4-b96933ea10b5","added_by":"auto","created_at":"2025-03-24 06:09:36","extension":"docx","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":143235,"visible":true,"origin":"","legend":"","description":"","filename":"TOC.docx","url":"https://assets-eu.researchsquare.com/files/rs-6201748/v1/62a7e91a676b72941e31649f.docx"},{"id":79071370,"identity":"ea31c1fe-92de-45e2-be35-2d22679ff442","added_by":"auto","created_at":"2025-03-24 06:09:35","extension":"jpg","order_by":2,"title":"","display":"","copyAsset":false,"role":"supplement","size":120379,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eScheme 1. \u003c/strong\u003eSchematic Diagram of the Preparation Process of HBP Wound Dressing\u003c/p\u003e","description":"","filename":"Scheme1.jpg","url":"https://assets-eu.researchsquare.com/files/rs-6201748/v1/af3471e017b90382d122b76f.jpg"}],"financialInterests":"No competing interests reported.","formattedTitle":"\u003cp\u003eStudy on the Preparation of Multifunctional Bacterial Cellulose-Based Wound Dressings and Their In Vitro Hemostatic Performance\u003c/p\u003e","fulltext":[{"header":"Introduction","content":"\u003cp\u003eUncontrollable bleeding is a primary cause of death following trauma, often leading to rapid mortality and complications such as coagulopathy, hypothermia, and severe infection\u003csup\u003e\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e,\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e\u003c/sup\u003e. Current commercial hemostatic materials, commonly used in military settings, are composed of polysaccharides, silicoaluminates, biological products, and peptides\u003csup\u003e\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e\u003c/sup\u003e.However, these materials are often ineffective for moderate to severe bleeding, may cause side effects, and are relatively expensive\u003csup\u003e\u003cspan additionalcitationids=\"CR5\" citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e\u003c/sup\u003e. Given these limitations, there is a critical need for rapid and efficient hemostatic materials suitable for civilian use, highlighting the importance of research in this area.\u003c/p\u003e \u003cp\u003eChitosan quaternary ammonium salt (HACC, Scheme \u003cspan refid=\"Sch1\" class=\"InternalRef\"\u003e1a\u003c/span\u003e) is prized for its excellent water solubility, flocculation, biocompatibility, hygroscopicity, moisture retention, and antibacterial properties, making it a versatile material in biomaterials research\u003csup\u003e\u003cspan additionalcitationids=\"CR8\" citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e\u003c/sup\u003e. For instance, Min et al.\u003csup\u003e\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e\u003c/sup\u003e HACC/PVA composite coating modified with quaternary ammonium salt via an eco-friendly solution casting method. It achieves 99% inhibition\u003c/p\u003e \u003cp\u003eagainst E. coli and S. aureus, and features transparency, biodegradability, anti-fogging, and antibacterial properties. Similarly. Mahdi Rahimi et al.\u003csup\u003e\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e\u003c/sup\u003e developed a quaternized chitosan/nanosilver composite membrane with antimicrobial properties to reduce wound infections. Tests using MTT assay and DAPI staining showed good biocompatibility and antibacterial performance. These studies highlight the potential of HACC as an effective antibacterial agent in non-toxic skin dressings. Its cationic nature allows it to form a barrier that adsorbs and kills bacteria, thereby protecting the wound surface and promoting healing.\u003c/p\u003e \u003cp\u003eHydroxyapatite (HAP, Scheme \u003cspan refid=\"Sch1\" class=\"InternalRef\"\u003e1b\u003c/span\u003e) is recognized for its biocompatibility, non-toxicity, non-irritating nature, and lack of immunogenicity, making it resistant to causing aging, sensitization, or carcinogenesis \u003csup\u003e\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e\u003c/sup\u003e. It significantly contributes to enhancing osteoblast proliferation, bone regeneration, and hemostatic processes\u003csup\u003e\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e,\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e\u003c/sup\u003e. Emerging research indicates that materials based on HAP can engage with blood constituents to accelerate clotting\u003csup\u003e\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e,\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e\u003c/sup\u003e. For instance, Noh and colleagues engineered a composite of plant polysaccharides and HAP designed for fracture hemostasis, characterized by its high solubility, swift hemostatic action, and adhesive qualities\u003csup\u003e\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e\u003c/sup\u003e. Similarly, Tham et al. discovered that a composite material (BoneSeal\u0026reg;), which combines HAP with biodegradable polylactic acid, exhibits dual capabilities in bone restoration and hemostasis, effectively minimizing blood loss from fractures\u003csup\u003e\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e\u003c/sup\u003e. Consequently, mineral-based hemostatic materials utilizing HAP are emerging as a promising solution for external hemostasis and the facilitation of tissue repair.\u003c/p\u003e \u003cp\u003eBacterial cellulose (BC), produced by microorganisms such as Gluconacetobacter xylinus, is a natural nanofibrous material distinguished by its superior water retention, crystallinity, mechanical strength, and intricate three-dimensional network structure compared to plant-derived cellulose\u003csup\u003e\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e,\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e\u003c/sup\u003e. BC is highly biocompatible, biodegradable, non-toxic, and hypoallergenic, making it suitable for medical applications\u003csup\u003e\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e\u003c/sup\u003e. Its breathable nature minimizes the risk of wound infections, and it can be further functionalized to exhibit antimicrobial properties\u003csup\u003e\u003cspan additionalcitationids=\"CR23\" citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e\u003c/sup\u003e. The thickness, morphology, and functionality of BC can be precisely engineered, allowing its membranes to be customized for specific hemostatic requirements\u003csup\u003e\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e,\u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e\u003c/sup\u003e. Additionally, BC facilitates wound healing by maintaining a moist microenvironment conducive to tissue regeneration, positioning it as an excellent candidate for advanced wound care dressings in modern medicine \u003csup\u003e\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e,\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e \u003cp\u003eIn this study, HACC and HAP were integrated into the 3D network of BC (derived from kombucha-fermented pear juice) using soaking, stirring, and freeze-drying techniques, resulting in the development of HBP, a composite material with enhanced antibacterial and hemostatic properties.\u003c/p\u003e \u003cp\u003eThe study results show that the HBP wound dressing has excellent antibacterial properties, with clear inhibition zones against Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, and Streptococcus mutans. Additionally, HBP reduces the hemostatic time to 300\u0026thinsp;\u0026plusmn;\u0026thinsp;5 seconds by effectively concentrating coagulation factors and platelets, which significantly accelerates clotting compared to pure BC.\u003c/p\u003e"},{"header":"Materials and methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003eMaterials\u003c/h2\u003e \u003cp\u003eKombucha was purchased from Shandong Zhi's Gaibao Biotechnology Co, Ltd. Yeast extract was obtained from Beijing Aoboxing Biotechnology Co, Ltd. Glucose, chitosan quaternary ammonium salt (with a degree of substitution\u0026thinsp;\u0026gt;\u0026thinsp;98%), and hydroxyapatite (with a purity\u0026thinsp;\u0026ge;\u0026thinsp;97%) were purchased from Shandong Keyuan Biochemical Co, Ltd.\u003c/p\u003e \u003c/div\u003e\n\u003ch3\u003eMethods\u003c/h3\u003e\n\u003cp\u003ePreparation of BC Films\u003c/p\u003e \u003cp\u003eIn a 250 mL conical flask, 100 mL of pear juice supernatant was mixed with 6% glucose and 1% yeast extract. After high-temperature sterilization, 10 mL of kombucha inoculum was added, and the flask was sealed with sterilized gauze. The mixture was statically cultured at 30\u0026deg;C for 7 days. The resultant bacterial cellulose (BC) film was washed with distilled water, treated with 1 wt% NaOH for 24 hours to remove impurities, washed again, soaked in 0.5 wt% glacial acetic acid for 24 hours, and finally washed until neutral. The film was then freeze-dried to obtain the purified BC.\u003c/p\u003e \u003cp\u003ePreparation of HBC Antibacterial Film\u003c/p\u003e \u003cp\u003eBC was immersed in a 2.0% (w/v) HACC solution (100 mL) for 3 days, then washed with deionized water and freeze-dried to obtain the antibacterial BC-HACC film (HBC).\u003c/p\u003e \u003cp\u003ePreparation of the Multifunctional Antibacterial Wound Dressing (HBP)\u003c/p\u003e \u003cp\u003eDissolve 1.0 g of the material in 100 mL water, add HBC, and stir magnetically for 3 hours to adhere HAP. Rinse with deionized water and vacuum freeze-dry to obtain the multifunctional BC-based wound dressing (BC-HACC-HAP, named HBP). The technical route is shown in Scheme \u003cspan refid=\"Sch1\" class=\"InternalRef\"\u003e1\u003c/span\u003e.\u003c/p\u003e \u003cp\u003eX-ray Diffraction Analysis (XRD)\u003c/p\u003e \u003cp\u003eXRD tests on BC, HBC, and HAP composites were conducted using a Rigaku Smartlab 9kW diffractometer (Japan), with a scanning range of 5\u0026deg;-80\u0026deg;at 10\u0026deg;/min.\u003c/p\u003e \u003cp\u003eFourier-Transform Infrared Spectroscopy Analysis (FT-IR)\u003c/p\u003e \u003cp\u003eFT-IR analysis of BC, HBC, and HAP composites was conducted using a Thermo Scientific Nicolet iS20 spectrometer with ATR method, scanning 16 times over 400\u0026ndash;4000 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e.\u003c/p\u003e \u003cp\u003eX-ray Photoelectron Spectroscopy (XPS)\u003c/p\u003e \u003cp\u003eXPS analysis of BC, HBC, and HBP composites was conducted using a Shimadzu/Kratos AXIS Ultra DLD spectrometer to measure O, C, Cl, Ca, and P content.\u003c/p\u003e \u003cp\u003eScanning Electron Microscopy Analysis (SEM)\u003c/p\u003e \u003cp\u003eSEM analysis of BC, HBC, and HAP composites was performed using a Hitachi Regulus 8100 microscope at 10 kV, with EDS elemental analysis on selected areas. Sample size was 5 mm \u0026times; 1 mm.\u003c/p\u003e \u003cp\u003ePerformance Testing of the Material's Water Management System\u003c/p\u003e \u003cp\u003eWater Content Test: Water content was measured by weighing samples (M1), freeze-drying them, and reweighing (M2). Wwater Content was calculated using the formula:\u003cdiv id=\"Equa\" class=\"Equation\"\u003e\u003cdiv format=\"TEX\" class=\"mathdisplay\" id=\"FileID_Equa\" name=\"EquationSource\"\u003e\n$$\\:{W}_{\\text{w}\\text{a}\\text{t}\\text{e}\\text{r}\\:\\text{c}\\text{o}\\text{n}\\text{t}\\text{e}\\text{n}\\text{t}}=\\frac{{M}_{1}-{M}_{2}}{{M}_{3}}\\times\\:100\\%$$\u003c/div\u003e\u003c/div\u003e\u003c/p\u003e \u003cp\u003eRehydration Test: Samples were soaked in distilled water for 24 hours, surface moisture removed, and weighed (M3). Wrehydration Rate was calculated as:\u003cdiv id=\"Equb\" class=\"Equation\"\u003e\u003cdiv format=\"TEX\" class=\"mathdisplay\" id=\"FileID_Equb\" name=\"EquationSource\"\u003e\n$$\\:{W}_{\\text{r}\\text{e}\\text{h}\\text{y}\\text{d}\\text{r}\\text{a}\\text{t}\\text{i}\\text{o}\\text{n}}=\\frac{{M}_{3}-{M}_{2}}{{M}_{3}}\\times\\:100\\%$$\u003c/div\u003e\u003c/div\u003e\u003c/p\u003e \u003cp\u003eWater Vapor Permeability (WVT) Test: A 50 mL beaker with 10 mm distilled water was covered with the sample, sealed with a rubber band, and tested at 38\u0026deg;C, 50% humidity for 24 hours (effective area: 0.001963 m\u0026sup2;). A plastic-wrapped beaker served as control. WVT was calculated using the formula:\u003cdiv id=\"Equc\" class=\"Equation\"\u003e\u003cdiv format=\"TEX\" class=\"mathdisplay\" id=\"FileID_Equc\" name=\"EquationSource\"\u003e\n$$\\:WVT=\\frac{\\varDelta\\:m-\\varDelta\\:m{\\prime\\:}}{A\\bullet\\:t}$$\u003c/div\u003e\u003c/div\u003e\u003c/p\u003e \u003cp\u003eIn the formula: ∆m and ∆m₀are weight differences for the sample and blank, A is the effective area (m\u0026sup2;), and T is the test duration\u003c/p\u003e \u003cp\u003eThermogravimetric Analysis (TG)\u003c/p\u003e \u003cp\u003eTGA of BC, HBC, and HAP composites was performed using a HITACHI STA200 analyzer. Samples (20 mg) were heated from 30\u0026deg;C to 800\u0026deg;C at 20\u0026deg;C/min under nitrogen (50 L/min).\u003c/p\u003e \u003cp\u003eMechanical Property Testing\u003c/p\u003e \u003cp\u003eTensile tests on BC, HBC, and HAP composites were performed using an INSTRON 8872 universal testing machine. Specimens in the form of strips (10 mm \u0026times; 50 mm) were tested at a crosshead speed of 5 mm/min, with three replicates for each material to obtain average values.\u003c/p\u003e \u003cp\u003eIn Vitro Hemostasis Test\u003c/p\u003e \u003cp\u003e40 mg of BC, HBC, and HBP were each added to 2 mL of anticoagulated pig blood supplemented with 80 \u0026micro;L of 0.2 mol/L CaCl₂ to neutralize the anticoagulant. Blood coagulation time was measured using a silent mixer at 37\u0026deg;C, with a 45\u0026deg; oscillation and a rotation speed of 5 r. p. m. The anticoagulant used was 10% sodium citrate.\u003c/p\u003e \u003cp\u003eHemolysis Rate Test\u003c/p\u003e \u003cp\u003eBC, HBC, and HBP were added to 2 mL of 50\u0026times; diluted red blood cell suspension and incubated at 37\u0026deg;C for 2 hours. After centrifugation (3000 r. p. m, 10 min), 100 \u0026micro;L supernatant absorbance was measured at 540 nm. Positive control: 0.2% TX-100; negative control: no material. Hemolysis rate was calculated as:\u003cdiv id=\"Equd\" class=\"Equation\"\u003e\u003cdiv format=\"TEX\" class=\"mathdisplay\" id=\"FileID_Equd\" name=\"EquationSource\"\u003e\n$$\\:\\text{H}\\text{e}\\text{m}\\text{o}\\text{l}\\text{y}\\text{s}\\text{i}\\text{s}\\:\\text{R}\\text{a}\\text{t}\\text{e}=\\frac{{A}_{s}-{A}_{n}}{{A}_{p}-{A}_{n}}$$\u003c/div\u003e\u003c/div\u003e\u003c/p\u003e \u003cp\u003eAs, Ap, and An represent the hemolysis rates of the experimental, positive, and negative control groups, respectively.\u003c/p\u003e \u003cp\u003eCytotoxicity Test\u003c/p\u003e \u003cp\u003eRAW264.7 macrophages were seeded in a 96-well plate and cultured until 80\u0026ndash;90% confluence. The medium was then replaced with 100 \u0026micro;L of sample extracts at concentrations of 0, 10, 20, 40, and 80 mg/mL. After 24 hours of incubation, the cells were treated with CCK-8 reagent, and absorbance was measured. Each sample was tested in triplicate, with a blank control included. The culture conditions were maintained at 30\u0026deg;C with 5% CO₂.\u003c/p\u003e \u003cp\u003eAntibacterial Performance Evaluation\u003c/p\u003e \u003cp\u003eE. coli, P. aeruginosa, and S. aureus cultures were adjusted to 10⁶ CFU/mL using sterile PBS. Aliquots of 100 \u0026micro;L of each bacterial suspension were spread onto LB agar plates. Sterilized samples were placed on the agar surface, and the plates were incubated at 37\u0026deg;C for 24 hours. The sizes of the inhibition zones were measured. Each sample was tested in triplicate, and the results were averaged. S. mutans was processed similarly but incubated for 48 hours, with data recorded accordingly.\u003c/p\u003e"},{"header":"Results and Discussion","content":"\u003cdiv id=\"Sec6\" class=\"Section2\"\u003e \u003ch2\u003eSurface morphology of HBP\u003c/h2\u003e \u003cp\u003eFigure \u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e1\u003c/span\u003eA shows that freeze-dried HBP appears more transparent than pure BC, with a white powdery precipitate on the surface\u003csup\u003e\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e,\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e\u003c/sup\u003e. The transparency is attributed to the bleaching effect of acetic acid during HBC soaking, while the precipitate results from HAP deposition during the composite process. Figure\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e1\u003c/span\u003eB reveals that HBP maintains a three-dimensional fibrous structure, with plate-like HAP crystals formed through mineralization. These crystals are relatively large, and some HAP further grows into spherical shapes, unevenly filling the cellulose network\u003csup\u003e\u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e31\u003c/span\u003e\u003c/sup\u003e. Higher-magnification SEM images (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e1\u003c/span\u003eC) show that these spherical structures are composed of plate-like HAP crystals.\u003c/p\u003e \u003c/div\u003e\n\u003ch3\u003eEds Analysis\u003c/h3\u003e\n\u003cp\u003eFigure \u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e2\u003c/span\u003ea shows the total distribution of elements in the EDS analysis. Figure\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e2\u003c/span\u003eb-f are the EDS distribution maps of C, O, Cl, P, and Ca elements, respectively. These results confirm the presence of chlorine (Cl, from HACC) and phosphorus (P) and calcium (Ca, from HAP) on the surface of HBP, indicating that both HACC and HAP have been successfully integrated into the HBP network\u003csup\u003e\u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e32\u003c/span\u003e,\u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e33\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e \u003cdiv id=\"Sec8\" class=\"Section2\"\u003e \u003ch2\u003eStructural analysis of HBP\u003c/h2\u003e \u003cp\u003eFigure \u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e3\u003c/span\u003eA displays the XRD patterns of the samples. HBC exhibits enhanced crystallinity compared to BC due to dehydration and molecular rearrangement, although no new peaks emerge (curves a and b). In contrast, HBP, which incorporates HAP, displays characteristic peaks of BC (14.6\u0026deg;, 17.7\u0026deg;, 22.7\u0026deg;) and additional crystal plane peaks of HAP (002), (211), (112), (300), (202), (312), (222), (213), and (304), consistent with the standard card (JCPDS No. 09-0432). The XRD results confirm the presence of HAP on the cellulose, corroborating the FTIR data (curve c)\u003csup\u003e\u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e34\u003c/span\u003e\u003c/sup\u003e. Figure\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e3\u003c/span\u003eB illustrates the characteristic FTIR peaks of BC: -OH (3340 cm⁻\u0026sup1;), C-H (2850\u0026ndash;2920 cm⁻\u0026sup1;), -CH₃ (1640 cm⁻\u0026sup1;), and cellulose (1309 cm⁻\u0026sup1;). HBC shows a slight shift at 2920 cm⁻\u0026sup1; and enhanced C-H peaks, indicating the incorporation of HACC into the BC structure. The FTIR spectrum of HBP (curve c) reveals C\u0026thinsp;=\u0026thinsp;O peaks at 1611 cm⁻\u0026sup1; and P-O peaks at 960 cm⁻\u0026sup1; and 1021 cm⁻\u0026sup1;, confirming the presence of PO₄\u0026sup3;⁻. Figure\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e3\u003c/span\u003eC and \u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e3\u003c/span\u003eD present XPS analysis, revealing the presence of Cl, P, and Ca elements in the material due to the incorporation of HACC and HAP. The binding energies at Cl 2p, P 2p, and Ca 2p further confirm their successful integration into the BC network\u003csup\u003e\u003cspan additionalcitationids=\"CR36\" citationid=\"CR35\" class=\"CitationRef\"\u003e35\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e37\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e \u003c/div\u003e\n\u003ch3\u003ePerformance Analysis of HBP\u003c/h3\u003e\n\u003cp\u003eFigure \u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e4\u003c/span\u003eA: DTG curves indicate weight loss between 38\u0026ndash;130\u0026deg;C due to moisture evaporation. HBC and HBP degrade at 246\u0026deg;C (attributed to HACC), while HAP raises the decomposition temperature to 429\u0026deg;C and increases residual mass, demonstrating superior thermal stability for HBP\u003csup\u003e\u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e38\u003c/span\u003e\u003c/sup\u003e. Figure\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e4\u003c/span\u003eB: Tensile tests reveal BC with a strength of 29.43\u0026thinsp;\u0026plusmn;\u0026thinsp;0.71 MPa and modulus of 18.59\u0026thinsp;\u0026plusmn;\u0026thinsp;0.82 MPa. Incorporation of HACC and HAP reduces HBC's strength to 17.98\u0026thinsp;\u0026plusmn;\u0026thinsp;0.766 MPa and modulus to 10.27\u0026thinsp;\u0026plusmn;\u0026thinsp;1.06 MPa. However, HBP recovers to a strength of 26.19\u0026thinsp;\u0026plusmn;\u0026thinsp;2.24 MPa and modulus of 17.92\u0026thinsp;\u0026plusmn;\u0026thinsp;1.64 MPa, with HAP enhancing mechanical properties by tightening fiber bonds\u003csup\u003e\u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e39\u003c/span\u003e\u003c/sup\u003e. Figure\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e4\u003c/span\u003eC: In vitro hemostasis tests show that HBP achieves the fastest clotting time (300\u0026thinsp;\u0026plusmn;\u0026thinsp;5 s) compared to others (582\u0026thinsp;\u0026plusmn;\u0026thinsp;4 s, 484\u0026thinsp;\u0026plusmn;\u0026thinsp;8 s, 513\u0026thinsp;\u0026plusmn;\u0026thinsp;10 s). HBP's high water absorption and 3D structure concentrate coagulation factors, while HAP releases Ca\u0026sup2;⁺ to further promote clotting\u003csup\u003e\u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e40\u003c/span\u003e,\u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e41\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e \u003cp\u003eTable\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e presents the water management performance of the samples. BC demonstrated excellent water retention and moisture permeability, with a water content of 98.02\u0026thinsp;\u0026plusmn;\u0026thinsp;0.10%, rehydration rate of 87.35\u0026thinsp;\u0026plusmn;\u0026thinsp;0.58%, and water vapor transmission rate (WVTR) of 5335.84\u0026thinsp;\u0026plusmn;\u0026thinsp;2.39 g/m\u0026sup2;\u0026middot;24h. These properties are attributed to its high surface area and porosity. HBC and HBP exhibited slightly lower water retention and rehydration rates (98.82\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03%, 84.58\u0026thinsp;\u0026plusmn;\u0026thinsp;0.63% for HBC; 96.22\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02%, 82.16\u0026thinsp;\u0026plusmn;\u0026thinsp;0.46% for HBP), but maintained good moisture permeability (6106.76\u0026thinsp;\u0026plusmn;\u0026thinsp;2.88 g/m\u0026sup2;\u0026middot;24h for HBC; 5528.36\u0026thinsp;\u0026plusmn;\u0026thinsp;4.12 g/m\u0026sup2;\u0026middot;24h for HBP). Despite the influence of composite materials, HBP still provided an effective sterile healing environment for wound repair due to its high moisture permeability.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eWater Content, Rehydration Rate, and Water Vapor Transmission Rate (WVT) of BC, HBC, and HBP\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"7\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c3\" namest=\"c2\"\u003e \u003cp\u003edry weight(g)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c5\" namest=\"c4\"\u003e \u003cp\u003ewater conten(%)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003erehydratio(%)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e \u003cp\u003ewvt(g/m2*24h)\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c2\" namest=\"c1\"\u003e \u003cp\u003eBC\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.88\u0026thinsp;\u0026plusmn;\u0026thinsp;0.21\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e98.02\u0026thinsp;\u0026plusmn;\u0026thinsp;0.10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c6\" namest=\"c5\"\u003e \u003cp\u003e87.35\u0026thinsp;\u0026plusmn;\u0026thinsp;0.58\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e5335.84\u0026thinsp;\u0026plusmn;\u0026thinsp;2.39\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c2\" namest=\"c1\"\u003e \u003cp\u003eHBC\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.84\u0026thinsp;\u0026plusmn;\u0026thinsp;0.30\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e98.82\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c6\" namest=\"c5\"\u003e \u003cp\u003e84.58\u0026thinsp;\u0026plusmn;\u0026thinsp;0.63\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e6106.76\u0026thinsp;\u0026plusmn;\u0026thinsp;2.88\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c2\" namest=\"c1\"\u003e \u003cp\u003eHBP\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.87\u0026thinsp;\u0026plusmn;\u0026thinsp;0.43\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e96.22\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c6\" namest=\"c5\"\u003e \u003cp\u003e82.16\u0026thinsp;\u0026plusmn;\u0026thinsp;0.46\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e5528.36\u0026thinsp;\u0026plusmn;\u0026thinsp;4.12\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e\n\u003ch3\u003eBiocompatibility Analysis of HBP\u003c/h3\u003e\n\u003cp\u003eFigure \u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e5\u003c/span\u003e illustrates the biocompatibility of the sample membranes. Figure\u0026nbsp;\u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e5\u003c/span\u003eA shows that BC, HBC, and HBP have hemolysis rates of 1.61%, 1.86%, and 4.24%, respectively, all below the 5% threshold for hemocompatibility\u003csup\u003e\u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e42\u003c/span\u003e\u003c/sup\u003e. This indicates that the incorporation of HACC and HAP does not damage red blood cells, which remain healthy and evenly distributed, making these materials suitable for biomedical applications involving blood contact. Figure\u0026nbsp;\u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e5\u003c/span\u003eB demonstrates that HBP exhibits good biocompatibility, with high cell survival rates for RAW264.7 macrophages across all extract concentrations, even slightly promoting cell growth. Thus, HBP shows excellent hemocompatibility and no cytotoxicity within the 20\u0026ndash;80 mg/mL range.\u003c/p\u003e \u003cdiv id=\"Sec11\" class=\"Section2\"\u003e \u003ch2\u003eAntibacterial Analysis of HBP\u003c/h2\u003e \u003cp\u003eFigure \u003cspan refid=\"Fig7\" class=\"InternalRef\"\u003e6\u003c/span\u003e presents the antibacterial performance of BC, HBC, and HBP against four common pathogens: Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, and Streptococcus mutans. BC exhibited no antibacterial activity, while HBC and HBP demonstrated clear inhibition zones, indicating strong antibacterial effects. HBP showed superior performance due to the slow release of Ca\u0026sup2;⁺ and PO₄\u0026sup3;⁻ from HAP, which enhances local ion concentration and boosts antibacterial efficacy\u003csup\u003e\u003cspan citationid=\"CR43\" class=\"CitationRef\"\u003e43\u003c/span\u003e,\u003cspan citationid=\"CR44\" class=\"CitationRef\"\u003e44\u003c/span\u003e\u003c/sup\u003e. Thus, HBP effectively prevents bacterial infections when used as a hemostatic dressing.\u003c/p\u003e \u003c/div\u003e"},{"header":"Conclusion","content":"\u003cp\u003eThis study developed an HBP composite dressing with antibacterial and hemostatic properties by incorporating HACC and HAP into BC's 3D network through soaking, stirring, and freeze-drying. The HBP dressing exhibits no cytotoxicity, meets hemolysis standards, and demonstrates improved thermal stability and tensile strength, making it suitable for medical applications. Compared to BC, HBP effectively prevents bacterial infections during hemostasis and achieves a hemostatic time of 300\u0026thinsp;\u0026plusmn;\u0026thinsp;5 seconds by promoting blood cell aggregation and activating coagulation at the wound site. Overall, HBP shows potential as a novel multifunctional hemostatic dressing for preventing infections and achieving rapid hemostasis.\u003c/p\u003e "},{"header":"Declarations","content":"\u003cp\u003e \u003ch2\u003eCompeting interests\u003c/h2\u003e \u003cp\u003eThe authors declare no competing interests.\u003c/p\u003e \u003c/p\u003e\u003ch2\u003eAuthor Contribution\u003c/h2\u003e\u003cp\u003eMiaoyi Tian: Data curation, Investigation, Visualization, Writing -original draft, Writing-review \u0026amp; editing. Shengbiao Ma: Data curation, Investigation, Visualization, Writing -original draft. Zhenchao Dong: Investigation, Writing-review \u0026amp; editing. Shenao Wang: Investigation. Xinyu Liu: Investigation. Binhui Liu: Investigation. Huaizhi Cui: Investigation. Mingshuai Ma: Investigation. Hang Zheng: Investigation. Xin Liu: Conceptualization, Formal analysis, Funding acquisition, Methodology, Resources.\u003c/p\u003e\u003ch2\u003eAcknowledgement\u003c/h2\u003e\u003cp\u003eThis work was supported by the Key Research and Development (R\u0026amp;D) Projects in Heilongjiang Province, China (Project No. GA21C013)\u003c/p\u003e\u003ch2\u003eData Availability\u003c/h2\u003e\u003cp\u003eAll data generated or analysed during this study are included in this published article and its supplementary information files.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eLippi, G., Favaloro, E. J. \u0026amp; Cervellin, G. in Seminars in thrombosis and hemostasis. 083\u0026ndash;093 (Thieme Medical Publishers).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eRossaint, R. et al. Management of bleeding following major trauma: an updated European guideline. \u003cem\u003eCrit. 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Evaluation of the antibacterial properties and in-vitro cell compatibilities of doped copper oxide/hydroxyapatite composites. \u003cem\u003eColloids Surf., B\u003c/em\u003e. \u003cb\u003e209\u003c/b\u003e, 112194 (2022).\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"},{"header":"Scheme 1","content":"\u003cp\u003eScheme 1 is available in the Supplementary Files section.\u003c/p\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"scientific-reports","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"scirep","sideBox":"Learn more about [Scientific Reports](http://www.nature.com/srep/)","snPcode":"","submissionUrl":"","title":"Scientific Reports","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Scientific Reports","inReviewEnabled":true,"inReviewRevisionsEnabled":true},"keywords":"Bacterial cellulose, Chitosan quaternary ammonium salt, Hydroxyapatite, Antibacterial test, In vitro hemostasis","lastPublishedDoi":"10.21203/rs.3.rs-6201748/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-6201748/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eBacterial cellulose (BC) is a favored material for medical wound dressings due to its biocompatibility and water-holding capacity, yet it lacks inherent antibacterial and hemostatic capabilities. To enhance its functionality, a novel BC-based wound dressing (HBP) was engineered by integrating chitosan quaternary ammonium salt (HACC) and hydroxyapatite (HAP) into BC's 3D structure via soaking, stirring, and freeze-drying techniques. This composite dressing demonstrates significant antibacterial activity, evidenced by distinct inhibition zones in antimicrobial assays, and rapid hemostatic performance, with an in vitro clotting time of 300\u0026thinsp;\u0026plusmn;\u0026thinsp;5 seconds. Characterized using SEM, XRD, FT-IR, and XPS, HBP exhibits superior physicochemical characteristics and biocompatibility, positioning it as a highly viable option for advanced biomedical wound dressings.\u003c/p\u003e","manuscriptTitle":"Study on the Preparation of Multifunctional Bacterial Cellulose-Based Wound Dressings and Their In Vitro Hemostatic Performance","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-03-24 06:09:30","doi":"10.21203/rs.3.rs-6201748/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision requested","date":"2025-05-16T12:15:26+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-05-16T07:14:08+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"114822545436722243919183617668303217415","date":"2025-04-28T04:22:23+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-04-09T18:59:18+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"115563036106474058258125166085067205576","date":"2025-04-04T05:59:00+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"35253988368250418121910044199223707794","date":"2025-03-31T05:37:25+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2025-03-21T00:19:07+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2025-03-21T00:13:58+00:00","index":"","fulltext":""},{"type":"editorInvited","content":"","date":"2025-03-20T15:18:49+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2025-03-20T06:27:53+00:00","index":"","fulltext":""},{"type":"submitted","content":"Scientific Reports","date":"2025-03-11T09:27:14+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"scientific-reports","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"scirep","sideBox":"Learn more about [Scientific Reports](http://www.nature.com/srep/)","snPcode":"","submissionUrl":"","title":"Scientific Reports","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Scientific Reports","inReviewEnabled":true,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"ebdccb0f-035e-491e-827c-25c7a122b6d7","owner":[],"postedDate":"March 24th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[{"id":46100080,"name":"Physical sciences/Materials science/Biomaterials"},{"id":46100081,"name":"Biological sciences/Microbiology/Antimicrobials"}],"tags":[],"updatedAt":"2026-04-27T16:06:53+00:00","versionOfRecord":{"articleIdentity":"rs-6201748","link":"https://doi.org/10.1038/s41598-026-48080-7","journal":{"identity":"scientific-reports","isVorOnly":false,"title":"Scientific Reports"},"publishedOn":"2026-04-22 15:58:56","publishedOnDateReadable":"April 22nd, 2026"},"versionCreatedAt":"2025-03-24 06:09:30","video":"","vorDoi":"10.1038/s41598-026-48080-7","vorDoiUrl":"https://doi.org/10.1038/s41598-026-48080-7","workflowStages":[]},"version":"v1","identity":"rs-6201748","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-6201748","identity":"rs-6201748","version":["v1"]},"buildId":"XKTyCvWXoU3ODBz1xrDgd","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}