A ZnO@ZIF-8/Ag Dual-Platform Nanocomposite Synergizing ROS Generation for Bactericidal-Immunomodulatory Wound Therapy

A ZnO@ZIF-8/Ag Dual-Platform Nanocomposite Synergizing ROS Generation for Bactericidal-Immunomodulatory Wound Therapy | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article A ZnO@ZIF-8/Ag Dual-Platform Nanocomposite Synergizing ROS Generation for Bactericidal-Immunomodulatory Wound Therapy Ruiling Hu, Yujie Zhang, Chengzhi Gao, Hang Wei, Xianjian Hong, and 8 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-7961947/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract The treatment of wounds caused by drug-resistant bacteria is a significant challenge in clinical medicine. Silver (Ag) nanoparticles have attracted considerable attention due to their ability to inhibit drug-resistant bacteria. Nevertheless, the propensity of Ag nanoparticles to aggregate as well as their elevated toxicity have restricted their practical application. To address this issue, this study involved the design of a core-shell-type ZnO@ZIF-8/Ag nanocomposite material that combines high antibacterial activity with excellent biocompatibility. The mean diameter of the Ag nanoparticles in this material was approximately 2.4 nm, and they were highly dispersed. Within the wound microenvironment medium, antibacterial factors, such as hydroxyl radicals (·OH), singlet oxygen ( 1 O 2 ), Zn 2+ , and Ag + , were generated. The material induced bacterial death by altering the secondary structure of the cell wall of drug-resistant bacteria, thereby inhibiting respiration, lysing phospholipid layers, causing cellular content leakage, and disrupting β -lactamases. The introduction of zinc oxide (ZnO) significantly reduced the toxicity of the Ag nanoparticles and regulated macrophage polarization, which inhibited the secretion of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) by M1-type macrophages (M1) while concomitantly promoting the secretion of interleukin-10 (IL-10) and vascular endothelial growth factor (VEGF) by M2-type macrophages (M2). The expression of platelet-endothelial cell adhesion molecule-1 (CD31), type I collagen/fibronectin (COL-Ⅰ/FN), and proliferating cell nuclear antigen (PCNA) was significantly promoted, which significantly enhanced wound healing in infected wounds. This study thus offers a novel strategy for developing therapies against drug-resistant bacterial infections with the potential for clinical application. Zinc-silver composite material Antibacterial Antibacterial mechanism Wound healing Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Figure 7 Figure 8 1 Introduction The issue of antibiotic-resistant bacterial infections caused by antibiotic misuse and overuse has become a significant challenge to human health [ 1 , 2 ]. According to a study published in The Lancet , over the course of the next 25 years, the number of fatalities resulting from antibiotic-resistant bacterial infections may exceed 39.1 million globally [ 3 ]. Consequently, an urgent need exists for the development of new antimicrobial strategies to address this growing threat [ 4 ]. Silver nanoparticles (Ag NPs) are regarded as a promising alternative to combat the issue of antibiotic resistance owing to their broad-spectrum antimicrobial properties [ 5 ]. The efficacy of Ag NPs in eliminating pathogens is attributable to multiple bactericidal mechanisms, including the release of metal ions, which disrupt bacterial membrane structures [ 6 ], the generation of reactive oxygen species (ROS), which interfere with DNA transcription and translation [ 7 ], and the synergy of metal ions with ROS to inactivate key enzymes [ 8 ]. This multifaceted approach significantly reduces the risk of antibiotic resistance while ensuring the efficient elimination of pathogenic bacteria. However, the antibacterial activity of Ag NPs is size-dependent, with smaller particles exhibiting stronger antibacterial performance. Notwithstanding, it has been demonstrated that larger specific surface areas increase the likelihood of aggregation and that Ag NPs tend to accumulate in biological organelles, which increases the risk of toxicity [ 9 ]. Research has shown that when NP sizes are less than 5 nm, they can be excreted safely from the body via the glomerular filtration pores [ 10 , 11 ]. The development of antimicrobial agents comprising ultra-small Ag NPs that exhibit both good dispersibility and excellent biosafety has thus become a significant challenge for the application of Ag NPs in the medical field. Zinc (Zn) is an essential trace element for the human body. It is involved in the composition and activation of various enzymes within the body and has been shown to play a crucial role in immune system repair [ 12 ]. Meanwhile, zinc oxide (ZnO), which has antibacterial properties and excellent biocompatibility, serves as an ideal drug carrier in the biomedical field [ 13 ]. However, when confronted with chronic infection wounds, ZnO appears to demonstrate a limited capacity to effectively inhibit bacterial growth and reproduction [ 14 ]. Theoretically, the loading of Ag NPs onto the ZnO surface has the potential to address the issue of Ag agglomeration and enhance biocompatibility. However, the presence of negative charges on both Ag and ZnO surfaces has been shown to facilitate the detachment of Ag NPs. Zeolite imidazolium framework material-8 (ZIF-8) is an emerging porous crystalline material. It is widely used as a medium in biomaterials due to its abundant active sites, high biocompatibility, strong antibacterial properties, and cell repair promotion capabilities [ 15 ]. Furthermore, the imidazole framework contains abundant nitrogen atoms, which possess lone pair electrons capable of forming coordinate bonds with electron-donating atoms or groups (the d electron orbitals of Ag NPs are empty). It can thus be concluded that the 2-methylimidazole (2-MIM) present within ZIF-8 stabilizes the loading of Ag NPs on the surface of ZnO by utilizing coordination bonds and Zn 2+ as active sites. Furthermore, the nitrogen elements within ZIF-8 firmly anchor the Ag NPs [ 16 – 18 ]. For the purpose of this study, ZIF-8 was used as an intermediary to synthesize core-shell-type ZnO@ZIF-8/Ag NPs. This was achieved by adsorbing Ag NPs with a particle size of approximately 2.4 nm onto the surface of a ZnO carrier (Fig. 1 A). The use of this strategy not only addressed the issue of Ag NP aggregation but also significantly reduced toxicity through the incorporation of ZnO and ZIF-8. The material generated ROS and metal ions in the microenvironment of infected wounds, which resulted in the bacterial cell wall and membrane integrity being compromised and the leakage of bacterial cytoplasm and nucleic acids, thereby ultimately leading to bacterial death. The experimental results, both in vitro and in vivo, validated the material’s high level of biocompatibility, which promoted the polarization of macrophages from M1 to M2 [ 19 , 20 ] and significantly enhanced wound healing in infected wounds (Fig. 1 B). Consequently, this study provides a theoretical foundation for the development of novel anti-infective bioactive materials and offers a novel solution to the issue of antibiotic resistance. 2 Results and Discussion 2.1 Preparation and Characterization of ZnO@ZIF-8/Ag NPs In this study, ZnO@ZIF-8/Ag NPs were prepared using a layer-by-layer assembly method. Initially, zinc acetate dihydrate (Zn (CH 3 COO) 2 ·2H 2 O) underwent reduction to ZnO NPs through a solvothermal process, in which diethylene glycol (DEG) was used as both the solvent and reducing agent. In the second experiment, Zn 2+ were used as a carrier to coordinate with 2-MIM to achieve the uniform growth of ZIF-8 on its surface. Finally, Ag sol with a particle size of approximately 2.4 nm was adsorbed onto the surface of the ZnO@ZIF-8 to obtain ZnO@ZIF-8/Ag NPs. Transmission electron microscopy (TEM) images revealed the particle sizes of the monodisperse ZnO (Fig. 2 A), ZnO@ZIF-8 (Fig. 2 B), and ZnO@ZIF-8/Ag (Fig. 2 C) as 358.8 ± 0.6, 364.6 ± 3.3, and 375.7 ± 2.8 nm (Fig. S1 ), respectively, with the particle size of the ZnO@ZIF-8/Ag/Ag measuring 2.4 ± 0.1 nm (Fig. S2). Conversely, the native Ag NPs had a propensity to agglomerate, with an average particle size of 42.4 ± 1.5 nm (Fig. S3). Energy-dispersive X-ray spectroscopy (EDS) maps (Fig. S4) and line scans (Fig. 2 D) demonstrated that the ZnO@ZIF-8/Ag NPs contained C, N, O, Zn, and Ag elements, the respective content of which was 53.42%, 7.11%, 23.95%, 14.15%, and 1.37%. The elemental mapping diagram (Fig. 2 E) illustrates the uniform distribution of the C (blue), N (yellow), O (red), Zn (green), and Ag (purple) on the ZnO@ZIF-8/Ag surface. X-ray diffraction (XRD) analysis of the material’s crystalline structure showed that the ZnO NPs exhibited a wurtzite phase (ICDD 36-1451) [ 21 ], while the Ag crystalline structure was face-centered cubic (ICDD 04-0783) [ 22 ] (Fig. 2 F). The characteristic peaks of ZIF-8 in Fig. 2 G, including the C-H stretching vibration (2929 cm − 1 ), the C = N stretching vibration in the imidazole ring (1583 cm − 1 ), and the C-N stretching vibrations (1145 cm − 1 and 994 cm − 1 ), indicated that the ZIF-8 successfully coordinated with the ZnO [ 15 ]. Concurrently, the ultraviolet-visible (UV-vis) molecular absorption spectrum (Fig. 2 H) of ZnO@ZIF-8 exhibited the characteristic absorption peak of ZIF-8 at 240 nm, while after loading the Ag NPs, the local surface plasmon resonance characteristic peaks of the Ag NPs were observed in the wavelength range 400–450 nm, which indicated a significant red shift and broadening of the spectral band [ 23 ]. This confirmed the successful loading of the Ag NPs onto the surfaces of the ZnO@ZIF-8 NPs. X-ray photoelectron spectroscopy (XPS) was utilized to investigate the elemental composition and valence states of the ZnO@ZIF-8/Ag. As demonstrated in Fig. 2 I, ZnO@ZIF-8/Ag contained the elements C, N, O, Zn, and Ag. The C 1s spectrum (Fig. 2 J) was decomposed into O-C = O (binding energy [BE] = 288.0 eV), C-OH (BE = 286.3 eV), and C-C (BE = 284.8 eV) [ 24 ]. The peaks at 400.9 eV, 399.8 eV, and 398.7 eV in the N 1s spectrum (Fig. 2 K) corresponded to N in the imidazole ring [ 25 ]. The O 1s spectrum (Fig. 2 L) was fitted within 530.5 and 531.6 eV, which corresponded to the lattice and surface oxygen within the ZnO microspheres. The XPS peaks at 1044.5 and 1021.5 eV were attributed to Zn 2p in the ZnO (Fig. 2 M) [ 26 ]. The XPS spectrum of Ag 3d was assigned to the Ag 3d 3/2 and Ag 3d 5/2 peaks at 373.8 and 367.8 eV (Fig. 2 N) [ 23 ]. 2.2 Mechanism of ROS Generation by ZnO@ZIF-8/Ag NPs Nano-Ag exhibits significant local surface plasmon resonance effects, which induce thermal electron escape during the plasmon resonance process. These high-energy electrons have the capacity to react with oxygen-containing media, and this reaction generates ROS. Furthermore, an established correlation exists between the electron transfer rate and the ROS production rate. Accordingly, under normal conditions, the rate of electron transfer is directly proportional to the level of ROS yield [ 27 ]. Consequently, electrochemical impedance spectroscopy (EIS) measurements were conducted in this study using a DH7000C electrochemical workstation, and Nyquist plots were generated [ 28 – 30 ]. In a Nyquist plot, a smaller semicircle radius is indicative of lower interfacial resistance and higher electron migration efficiency [ 31 ]. As shown in the Nyquist plot in Fig. 3 B, the radius of ZnO@ZIF-8/Ag is reduced, which suggests enhanced charge transfer efficiency. Density functional theory calculations were then utilized to further investigate the ROS generation process [ 32 ]. In an oxygen-rich environment, the ZnO@ZIF-8/Ag NPs interacted with the microenvironment of the infected wound to generate H 2 O 2 , and Ag promoted the breaking of the O-O bonds by providing electrons to produce ·OH. Under visible light, surface plasmon resonance occurred on the Ag NP surface, thereby overcoming the potential energy barrier to form 1 O 2 (Fig. 3 C) [ 33 , 34 ]. The experimental results demonstrated that Ag can efficiently adsorb H 2 O 2 (Fig. S5) and catalyze the generation of ROS on the Ag surface. The following equation was postulated: Ag + O 2 → Ag + + 1 O 2 Ag + H 2 O 2 → Ag + + ·OH + OH – To investigate the types of ROS generated by the ZnO@ZIF-8/Ag NPs, the following reagents were employed for validation: potassium permanganate (KMnO 4 ) [ 35 ], 3,3',5,5'-tetramethylbenzidine (TMB) [ 36 ] and 1,3-diphenylisobenzofuran (DPB) [ 37 ] (Fig. 3 D). In comparison with the control group (pure potassium permanganate), the ZnO NP, ZnO@ZIF-8 NP, and ZnO@ZIF-8/Ag NP groups all exhibited a decrease in their respective peak absorbances. The most significant decrease was observed in the ZnO@ZIF-8/Ag NP group, which suggested that this group had a superior H 2 O 2 generation capacity (Fig. 3 E). Subsequent analysis of the ·OH generation capacity (Fig. 3 F) demonstrated that the ZnO@ZIF-8/Ag NP group exhibited significantly higher levels of absorption compared to the other groups, which confirmed the effective catalysis of ·OH generation by this material. Furthermore, the 1,3-diphenylisobenzofuran (DPBF) degradation experiment (Fig. 3 G) demonstrated that the ZnO@ZIF-8/Ag NP group had the most rapid degradation rate, which indicated that this system generated a greater quantity of 1 O 2 .The results were further validated by electron paramagnetic resonance (EPR) spectroscopy, which confirmed that the 1:2:2:1 quadruplet peak was a characteristic absorption peak of ·OH (Fig. 3 H). In contrast, the single triplet peak at 1:1:1 corresponded to the characteristic peak of 1 O 2 (Fig. 3 I) [ 38 ], thus confirming that the ZnO@ZIF-8/Ag NPs possessed excellent synergistic ROS (H 2 O 2 , ·OH, and 1 O 2 ). In the event of bacterial infections manifesting at local wound sites, macrophages undergo M1-type polarization (pro-inflammatory phenotype) and secrete elevated levels of proinflammatory factors (e.g., TNF-α, IL-6) to facilitate the elimination of pathogens and damaged cells. Concurrently, the body upregulates glutathione (GSH) synthesis to neutralize the oxidative stress and protect the surrounding normal tissues from damage caused by the excessive inflammatory responses [ 39 ]. However, it has been demonstrated that excessive GSH can be utilized by bacteria to counteract the ROS produced by cells [ 40 ]. Further, in the context of antimicrobial experiments, the depletion of excessive GSH has been shown to enhance the efficacy of antimicrobials by reducing bacterial tolerance to ROS [ 35 , 41 ]. As depicted in Fig. 3 J, with an increase in the concentration of ZnO@ZIF-8/Ag NPs, the characteristic absorption peak of GSH at 412 nm declined. It is notable that at a concentration of 50 µg/mL of ZnO@ZIF-8/Ag NPs, the absorption peak underwent a substantial decline. This phenomenon strongly confirms that the ZnO@ZIF-8/Ag NP composite material exhibited significant activity in consuming GSH, which indicates its ability to efficiently catalyze the oxidation of GSH and thereby potentially induce oxidative stress in biological systems. 2.3 In vitro antibacterial activity testing of ZnO@ZIF-8/Ag NPs Upon oxygen adsorption, the ZnO@ZIF-8/Ag NPs generated ROS through interactions with wound microenvironment components. Theoretically, these highly oxidative ROS may have effectively oxidized the bacterial cell walls and disrupted the phospholipid bilayers of the cell membranes, which resulted in membrane perforation. Concurrently, the release of Zn 2+ and silver ions (Ag + ) enabled electrostatic binding to bacterial cell walls, thereby inducing structural alterations, which led to bacterial death [ 8 ] (Fig. 4 A). Three clinically relevant pathogens were selected to evaluate the antibacterial efficacy: gram-negative Escherichia coli ( E. coli ), gram-positive Staphylococcus aureus ( S. aureus ), and drug-resistant Salmonella (DR- Salmonella ). A disk diffusion assay (Fig. S6) demonstrated that the ZnO and ZnO@ZIF-8 NPs exhibited relatively weak antibacterial activity. In contrast, the ZnO@ZIF-8/Ag NPs displayed broad-spectrum, concentration-dependent antibacterial effects, with significant inhibition against both E. coli and S. aureus and notable efficacy against DR- Salmonella . As illustrated in Fig. 4 B-D, the optimal antibacterial concentration was determined to be 50 µg/mL for the ZnO@ZIF-8/Ag NPs, and this was used in subsequent antibacterial testing. The colony counting results (Fig. 4 E-G) confirmed that the composite material achieved 99.9% bactericidal efficiency against all three tested strains within 50 min. Quantitative analysis (Fig. 4 H and I) revealed that at the optimal concentration, the ZnO@ZIF-8/Ag NPs exhibited bactericidal rates of 99.9 ± 0.2%, 99.7 ± 0.12%, and 97.5 ± 1.5% against E. coli , S. aureus , and DR- Salmonella , respectively, after 30 min of exposure. As shown in Fig. S7, the minimum inhibitory concentrations (MIC) of the ZnO@ZIF-8/Ag NPs against E. coli , S. aureus , and DR- Salmonella were determined to be 15.63 ± 2.3, 31.25 ± 0.1, and 31.25 ± 0.4 µg/mL, respectively. These results collectively demonstrated that the ZnO@ZIF-8/Ag NPs exhibited outstanding antibacterial performance across the three distinct evaluation methods (disk diffusion assay, colony counting, and MIC determination), which highlights the potential of ZnO@ZIF-8/Ag as a highly effective antimicrobial material. During the process of reproduction, bacteria form biofilms in suitable external environments. The formation of biofilms is a multi-stage dynamic process that typically includes the following steps: adhesion, where bacteria reversibly attach to solid surfaces through physical and chemical interactions, and colonization, where microorganisms form irreversible, robust bonds with surfaces through the secretion of extracellular polymers. As microorganisms proliferate and extracellular polymers accumulate further, the biofilm gradually matures to form a complex three-dimensional structure with bacteria inside, which exhibits high antibiotic resistance and metabolic diversity [ 36 ]. Biofilms provide an effective protective barrier for bacteria by shielding them from phagocytosis and neutralizing antimicrobial agents. This, in turn, safeguards bacteria from host immune defense attacks. To assess the anti-biofilm activity of the ZnO@ZIF-8/Ag NPs, crystal violet staining experiments were conducted using three representative bacterial strains (Fig. 4 J). The control group exhibited biofilms characterized by robust structural integrity, while the biofilm biomass in the treated group was found to be significantly diminished. Quantitative analysis (Fig. S8) confirmed that the ZnO@ZIF-8/Ag NPs significantly inhibited the formation of biofilms in E. coli and S. aureus and demonstrated some inhibitory effects on the formation of biofilms in DR- Salmonella . These findings pointed to the possibility of using ZnO@ZIF-8/Ag NPs as an effective strategy against biofilm-associated infections. The inhibitory effect could be attributed to the following mechanisms: the disruption of bacterial adhesion, interference with EPS production, and the exertion of direct bactericidal effects on the cells embedded within the biofilm [ 42 ]. 2.4 Research on antibacterial mechanisms The primary components of bacterial cell walls (e.g., phospholipid layers, teichoic acid, lipopolysaccharides) are rich in negatively charged functional groups, such as phosphate (PO 4 3– ), carboxyl (COO − ), and hydroxyl (OH − ) groups. These structures are imperative for preserving bacterial integrity and metabolic functions. Theoretically, the metal ions released by the ZnO@ZIF-8/Ag NPs may have disrupted the bacterial cell walls through electrostatic adsorption, while the ROS generated by the nanomaterials oxidized the phospholipid molecules, thereby leading to cell wall lysis [ 6 , 8 , 43 ] (Fig. 5 A). To validate this hypothesis, a series of experiments were designed with the aim of assessing the antibacterial mechanisms of the nanocomposites. The ion release experiments (Fig. S9) confirmed that the ZnO@ZIF-8/Ag NPs released 78.5 ± 0.4% Zn 2+ and 67.4 ± 0.1% Ag + . Scanning electron microscopy (SEM) was used to discern the effects of the material on the morphological alterations of the bacteria. In the control group, the bacteria exhibited smooth surfaces with no obvious cytoplasmic leakage. In the experimental group, the bacterial cell surfaces exhibited clear indications of damage, which manifested as wrinkling and denting. Among the tested organisms, the cell membranes of E. coli and S. aureus were the most severely compromised, and this damage was accompanied by substantial leakage of cytoplasmic content. Fluorescence inverted microscopy showed that the bacteria treated with ZnO@ZIF-8/Ag NPs exhibited red fluorescence. This finding suggests that the integrity of the cell membrane structure was compromised, which enabled PI to enter the cell interior and bind to nucleic acids. In contrast, the untreated control bacteria primarily exhibited green fluorescence, which indicated intact cell membrane structures and viable bacterial cells (Fig. 4 K) [ 7 ]. These results demonstrated that the ZnO@ZIF-8/Ag NPs exhibited highly effective broad-spectrum antibacterial activity against E. coli , S. aureus , and DR- Salmonella . It was hypothesized that alterations in the potential of bacterial cell walls may be indicative of changes in the surface charge of bacterial cell membranes. The Zeta potential values for E. coli , S. aureus , and DR- Salmonella were − 15.59 ± 1.0, − 14.83 ± 2.4, and − 14.35 ± 1.7 mV, respectively. Following a period of 5 min during to allow for their interaction with the ZnO@ZIF-8/Ag NPs, the Zeta potentials of the three bacteria increased to − 10.85 ± 2.2, − 11.91 ± 0.1, and − 12.64 ± 0.3 mV, respectively. After a further 40 min of interaction, these potentials increased further to − 4.23 ± 1.2, − 5.33 ± 0.8, and − 7.06 ± 1.9 mV, respectively (Fig. 5 B). The results obtained from this study indicate that ZnO@ZIF-8/Ag NPs have the capacity to significantly neutralize the negative charges on the surfaces of bacterial cell membranes. Furthermore, the neutralization effect was found to be time-dependent. The data pertaining to the leakage of bacterial nucleic acid and cytoplasm (Fig. 5 C) demonstrated that the nucleic acid leakage in the experimental group increased by 0.58 ± 0.1, 0.35 ± 2.9, and 0.21 ± 1.3 for E. coli , S. aureus , and DR- Salmonella , respectively. The ion leakage experiments (Fig. 5 D) revealed significant reductions in ionic content compared to the control group. In E. coli , S. aureus , and DR- Salmonella , respectively: K⁺ levels decreased by 6.72 ± 0.1, 4.71 ± 0.9, and 5.88 ± 0.1 mg/mL; Ca²⁺ levels decreased by 8.91 ± 0.3, 3.37 ± 1.2, and 3.85 ± 1.1 mg/mL; Mg²⁺ levels decreased by 0.23 ± 0.1, 0.55 ± 0.3, and 0.1 ± 0.05 mg/mL. These data indicate that ZnO@ZIF-8/Ag has the capacity to compromise the integrity of bacterial cell membranes, leading to the efflux of intracellular ions and nucleic acids. Furthermore, microcalorimetric analysis (Fig. 5 E) demonstrated the absence of an exponential growth phase in the bacterial growth curve following treatment with ZnO@ZIF-8/Ag, with the majority of the heat output exhibiting stabilization. This suggests that ZnO@ZIF-8/Ag may inhibit bacterial energy metabolism by disrupting the respiratory systems of bacteria, thereby exerting its antibacterial effect [ 44 ]. β -Lactamases have been identified as significant contributors to bacterial antibiotic resistance, as they facilitate bacterial survival and proliferation in environments exposed to antibiotics. This phenomenon represents a substantial challenge in the field of clinical antimicrobial therapy [ 45 ]. To assess the hypothesis that ZnO@ZIF-8/Ag NPs enhance antimicrobial efficacy by inhibiting β -lactamase activity, the binding affinities of Zn 2+ , ZIF-8 and Ag + with β -lactamase were calculated separately (Fig. 5 F). The Bes of Zn 2+ , ZIF-8, and Ag + with β -lactamase were − 0.82, − 5.03, and − 0.21 kcal/mol, respectively (Fig. S10), which indicated that the inhibitory capacity against β -lactamase was ZIF-8 > Zn 2+ > Ag + . These results demonstrated that the nanocomposite likely exerted synergistic antibacterial effects through multiple mechanisms, as schematically illustrated in Fig. 5 G. Bacterial surfaces typically carry negative charges, so this enabled strong electrostatic interactions between the surface components (e.g., carboxyl and phosphate groups) and the released Zn 2+ /Ag + ions from the ZnO@ZIF-8/Ag NPs. Concurrently, the unique physical structure of ZIF-8 induced mechanical damage to the bacterial membranes upon contact, which compromised the membrane integrity. This structural disruption significantly altered the membrane permeability, disrupted the osmotic balance and caused massive leakage of the intracellular components (e.g., proteins, nucleic acids, ions), and thereby severely interfered with the normal bacterial physiology. Furthermore, the ZnO@ZIF-8/Ag NPs exhibited catalytic activity, which facilitated the adsorption and subsequent conversion of environmental H 2 O 2 and O 2 into highly oxidative hydroxyl radicals (·OH) and singlet oxygen ( 1 O 2 ). These ROS inflicted oxidative damage on critical biomacromolecules within the bacterial cells. Simultaneously, the nanocomposite oxidized the intracellular reduced GSH to oxidized glutathione (GSSG), which amplified the ROS-mediated antibacterial effects and disrupted the cellular redox homeostasis. The released Ag + ions specifically bound to the thiol groups on the bacterial DNA, thereby inducing strand breaks and base damage that compromised the genetic integrity of the bacteria. Additionally, Ag + disrupted the ion channels in the bacterial membranes, which led to the leakage of essential ions (e.g., K⁺, Na⁺, Ca²⁺) and consequent loss of cellular function. In summary, the ZnO@ZIF-8/Ag NPs achieved potent antibacterial efficacy through a multifaceted mechanism involving (1) electrostatic adsorption, (2) physical membrane damage, (3) ROS generation, (4) enzymatic interference, (5) DNA damage, and (6) ion channel disruption [ 46 ]. 2.5 Biocompatibility of ZnO@ZIF-8/Ag NPs The biological safety of materials utilized in wound dressings must be a primary consideration during evaluations. In this study, the biocompatibility of the ZnO@ZIF-8/Ag NPs was systematically assessed through a range of in vitro and in vivo methods, including cell co-culture, blood compatibility, cell migration, and tissue toxicity lesions. In the experimental setup, RAW 264.7 macrophages were co-cultured with ZnO@ZIF-8/Ag NPs. To distinguish between the macrophages and the ZnO@ZIF-8/Ag NPs, the macrophages were labelled with a fluorescent tag constructed using lentivirus and appeared red. Meanwhile, the ZnO@ZIF-8/Ag NPs were labelled with CY5-polyethylene glycol-sulfhydryl (CY5-PEG-SH) to appear green. Following phagocytosis of the ZnO@ZIF-8/Ag NPs, the cells exhibited yellow fluorescence. The results (Fig. 6 A) demonstrated the effective uptake of the ZnO@ZIF-8/Ag NPs by the macrophages. As depicted in Fig. 6 B, hemocompatibility testing revealed that, within the concentration range that was examined, treatment with ZnO@ZIF-8/Ag NPs did not cause significant hemolysis. The observed hemolysis rates were all found to be below the internationally recognized safety threshold of 5% [ 47 ]. Subsequently, scratch wound healing experiments were conducted using L929 mouse fibroblast cells (Fig. 6 C and D). In comparison with the cells devoid of ZnO@ZIF-8/Ag NPs, the cell migration rate in the ZnO@ZIF-8/Ag NP-treated group was significantly increased, reaching 68.9% at 12 h and 92.6% at 24 h. Furthermore, a series of histopathological examinations using hematoxylin and eosin (H&E) staining were conducted on major organs (heart, liver, spleen, lungs, and kidneys) of Sprague-Dawley rats, along with in vivo toxicity assessments (Fig. 6 E). No overt pathological changes were observed in the cellular morphology and histological structure of these organs. The results of the in vivo muscle injection experiment showed that the experimental group exhibited mild local muscle fiber ruptures, with no obvious cellular swelling, degeneration, or necrosis. These alterations were also observed in normal skeletal muscle cells, which indicated that the experimental treatment had minimal interference with basic cellular physiological functions [ 48 ]. In summary, the experimental findings from both the in vitro and in vivo contexts demonstrated that the ZnO@ZIF-8/Ag NPs exhibited favorable biocompatibility and promoted cell proliferation. 2.6 Antimicrobial and wound healing properties in vivo Zinc is a trace element that is indispensable to the human body, as it plays a crucial role in fundamental life processes, such as DNA synthesis, protein metabolism, cell proliferation, and immune regulation. Research has demonstrated that when zinc is applied directly to wounds and the surrounding damaged skin, it promotes the clearance of necrotic tissue, reduces the risk of infection, alleviates inflammation, and activates epithelialization [ 49 ]. In this study, a full-thickness skin S. aureus infectious wound model was established in Sprague-Dawley rats to evaluate the in vivo therapeutic efficacy of ZnO@ZIF-8/Ag (Fig. 7 A and B). On the third day, the ZnO@ZIF-8/Ag group exhibited a 49.48 ± 3.9% healing rate, which increased to 93.23 ± 1.5% on the sixth day and 99.78 ± 1.7% on the ninth day when compared with the control, ZnO, and ZnO@ZIF-8 groups (Fig. 7 D). Furthermore, the wounds treated with ZnO@ZIF-8/Ag exhibited no bacterial presence after day 9 (Fig. 7 C and Fig. S11). Dynamic monitoring revealed no significant difference in body weight between the S. aureus -infected group and the control group (Fig. 7 E), which confirmed the significant advantages and application potential of ZnO@ZIF-8/Ag in the healing of infected wounds. To further assess the process of wound healing in infected wounds, wound sections were collected on days 3 and 7, and H&E staining and Masson’s trichrome staining were performed. As demonstrated in Fig. 7 F, during the acute phase post-trauma (day 3), the control group exhibited extensive infiltration of inflammatory cells (primarily neutrophils and macrophages) and a loosened tissue structure; in contrast, the ZnO@ZIF-8/Ag-treated group had significantly reduced inflammatory cell infiltration and better reserved tissue structural integrity. By the repair phase (day 7), residual inflammatory cells and minimal collagen deposition were still visible in the control group, whereas the ZnO@ZIF-8/Ag group exhibited nearly complete resolution of the inflammatory response, with newly formed epithelial tissue intact. The dermis layer had abundant vascularization and orderly collagen fibers, with significantly increased collagen deposition. The results indicated that the ZnO@ZIF-8/Ag NPs material exhibited significant advantages in inhibiting inflammatory responses, promoting epithelial formation, and enhancing collagen deposition. 2.7 Immunofluorescence The process of wound healing is categorized into four distinct stages: hemostasis, inflammation, proliferation, and remodeling [ 20 ]. During the inflammatory phase, M1 (marked by CD86) initiate the inflammatory response by secreting pro-inflammatory factors, such as IL-6 and TNF-α, to eliminate pathogens and necrotic tissue. M2 (marked by CD206) subsequently promote tissue repair and angiogenesis by secreting anti-inflammatory factors, such as IL-10, and stimulating the body to produce pro-repair factors, such as platelet-endothelial cell adhesion molecule-1 (CD31), vascular endothelial growth factor (VEGF), type I collagen/fibronectin (COL-Ⅰ/FN), and proliferating cell nuclear antigen (PCNA). These factors participate in intercellular interactions and signal transduction and collectively drive the transition of wounds from the inflammatory phase to the proliferative phase, thereby ensuring the smooth progression of the wound healing process [ 50 , 51 ]. In the immunofluorescence images in this study, the distribution of CD86 (Fig. 8 A), IL-6 (Fig. 8 B), and TNF-α (Fig. 8 C) was lower in the ZnO@ZIF-8/Ag group. The expression levels of CD206 (Fig. 8 D) and IL-10 (Fig. 8 E) were elevated in the ZnO, ZnO@ZIF-8, and ZnO@ZIF-8/Ag groups compared to the group without added material, but particularly in the ZnO@ZIF-8/Ag group. This finding indicated that ZnO@ZIF-8/Ag effectively promoted the polarization of M1 macrophages toward M2 macrophages, reduced the destructive effects of inflammatory factors on tissues, enhanced the role of M2 macrophages in tissue repair, and accelerated the wound healing process. In the context of angiogenesis, CD31 is a marker for vascular endothelial cells, while VEGF is a key player in the process, with its expression levels reflecting the status of angiogenesis. COL-Ⅰ/FN and PCNA are significant components of the extracellular matrix, they play pivotal roles in cell adhesion, proliferation, and migration and serve as key markers for tissue repair during wound healing [ 52 , 53 ]. In the ZnO@ZIF-8/Ag sample, the expression levels of CD31 (Fig. 8 F), VEGF (Fig. 8 G), COL-Ⅰ (Fig. 8 H), FN (Fig. 8 I), and PCNA (Fig. S12) were found to be significantly elevated in comparison to those in the ZnO and ZnO@ZIF-8 samples. It is clear from the experimental evidence that ZnO@ZIF-8/Ag significantly advances the inhibition of inflammatory responses, the promotion of tissue repair, the acceleration of angiogenesis, and the enhancement of extracellular matrix synthesis, all of which provides robust support for its application in wound healing. 3 Conclusion This study involved the use of ZnO as a precursor and employed a ZIF-8 coordination-mediated strategy to synthesize ZnO@ZIF-8/Ag nanocomposites loaded with ultra-small Ag sol (approximately 2.4 nm) with the aim of treating wounds caused by drug-resistant bacteria. These nanocomposites exhibited high antibacterial activity and biocompatibility and promoted the healing of infected wounds. The antibacterial activity of ZnO@ZIF-8/Ag can be attributed to the generation of ROS and the release of Zn 2+ and Ag + ions with the simultaneous inactivation of bacteria through the regulation of the redox balance in the microenvironment through the removal of excess GSH. The ZnO@ZIF-8/Ag NPs demonstrated high levels of biological activity, as indicated by both the in vitro and in vivo biocompatibility results. Moreover, this nanocomposite material induced M1 macrophages to polarize toward the M2 phenotype, thereby reducing inflammatory responses in the in vivo experiments, promoting collagen synthesis and deposition, speeding up angiogenesis and tissue remodeling, and ultimately accelerating wound healing rates and achieving comprehensive tissue repair. In the present study, a novel nano-composite material-based synergistic treatment strategy for bacterial infection wounds was developed. This strategy has significant application potential and provides a theoretical basis and experimental foundation for the design of novel antimicrobial wound dressings. 4 Experimental section For all the experiments, please see the Supporting Information. Declarations Supplementary Information The online version contains supplementary material available at Author contributions Ruiling Hu: Writing–original draft, Software, Methodology, Data curation. Yujie Zhang: Writing–original draft, Investigation, Formal analysis, Data curation. Chengzhi Gao: Writing–original draft, Software, Data curation. Hang Wei: Investigation, Data curation. Xianjian Hong: Formal analysis, Validation, Investigation. Tinghui Qiang: Validation, Software, Methodology, Formal analysis. Zhongshang Guo: Formal analysis, Data curation. Li Huang: Methodology, Investigation. Xiaoyun Lei: Software, Methodology, Investigation, Formal analysis. Caibin Zhao: Investigation, Software, Formal analysis, Data curation. Xiaohui Ji: Writing–review & editing, Methodology, Funding acquisition, Project administration. Hao Han: Writing–review & editing, Methodology. Shaobo Guo: Writing–review & editing, Supervision, Investigation, Funding acquisition, Conceptualization. Funding This work was supported by the Science and Technology Innovation Team Project of Shaanxi Province (2025RS-CXTD-040), Shaanxi Provincial Natural Science Foundation (24JK0376), Key Research and Development Program of Shaanxi (2024SF-YBXM-158) and Shaanxi Province Health and Wellness Scientific Research Innovation Capacity Enhancement Project (2024PT-16). Additional funding was provided by the Open Project Program of the State Key Laboratory of Qinba Biological Resources and Ecological Environment (SLGPT2019KF04-01). Data availability Data will be made available on request. Competing interests The authors declare no competing interests. References X. Wang, X.D. Qin, Y. Liu, Y.T. Fang, H. Meng, M.L. Shen, L.L. Liu, W.W. Huan, J. Tian, Y.W. Yang (2024) Plasmonic Supramolecular Nanozyme-Based Bio-Cockleburs for Synergistic Therapy of Infected Diabetic Wounds, Adv. Mater. 36(49):2411194. https://doi.org/10.1002/adma.202411194 A. Abbas, A. Barkhouse, D. Hackenberger, G.D. Wright (2024) Antibiotic resistance: A key microbial survival mechanism that threatens public health, Cell Host Microbe 32(6):837-851. https://doi.org/10.1016/j.chom.2024.05.015 M. Naghavi, S. E. Vollset, K. S. Ikuta, L. R. Swetschinski, A. P. Gray, E. E. Wool, et al., (2024) Global burden of bacterial antimicrobial resistance 1990–2021: a systematic analysis with forecasts to 2050, Lancet 404(10459):1199-1226. https://doi.org/10.1016/S0140-6736(24)01867-1 J. Song, Y.H. Lu, T. Pan, J.F. Wang, Z.Q. Liu, L. Xu, S.Q. Zhang, Y.P. Li, Y.Y. Bai, B.C. Heng, X.N. Zheng, H.Y. Wu, X.Y. Han, Y.S. Guo, Q. Cui, X.L. Deng, X.H. Zhang (2024) Manipulation of Surface Electrical Charge on Nanocomposite Membranes Confers Wide Spectrum Bactericidal Effects and Promotes Tissue Regeneration, Adv. Funct. Mater. 34(22):2314024. https://doi.org/10.1002/adfm.202314024 X.Y. Gong, N.D. Jadhav, V.V. Lonikar, A.N. Kulkarni, H.K. Zhang, B.R. Sankapal, J.N. Ren, B. Bin Xu, H.M. Pathan, Y. Ma, Z.P. Lin, E. Witherspoon, Z. Wang, Z.H. Guo (2024) An overview of green synthesized silver nanoparticles towards bioactive antibacterial, antimicrobial and antifungal applications, Adv. Colloid Interface Sci. 323:103053. https://doi.org/10.1016/j.cis.2023.103053 C.Y. Wang, S.L. Zhu, Y.Q. Liang, C.L. Qin, F. Wang, H. Wang, C.T. Chang (2023) A. Inoue, Flexible free-standing antibacterial nanoporous Ag ribbon, J. Colloid Interface Sci. 645:287-296. https://doi.org/10.1016/j.jcis.2023.04.153 Y.X. Wang, M.J. Malkmes, C. Jiang, P. Wang, L.Y. Zhu, H.M. Zhang, Y.H. Zhang, H. Huang, L. Jiang (2021) Antibacterial mechanism and transcriptome analysis of ultra-small gold nanoclusters as an alternative of harmful antibiotics against Gram-negative bacteria, J. Hazard. Mater. 416:126236. https://doi.org/10.1016/j.jhazmat.2021.126236 D.L. Han, X.M. Liu, S.L. Wu (2022) Metal organic framework-based antibacterial agents and their underlying mechanisms, Chem. Soc. Rev. 51(16):7138-7169. https://doi.org/10.1039/D2CS00460G Z. Li, T. Yang, X. Li, P. Yin, B. Yang, D. Li, Y. Wang, W. Teng, Q. Yu, W. Li (2025) A Bio-Responsive Hydrogel with Spatially Heterogeneous Structure for Treating Infectious Tissue Injuries, Adv Sci (Weinh) 12(23):2500088. https://doi.org/10.1002/advs.202570182 B. Du, X. Jiang, A. Das, Q. Zhou, M. Yu, R. Jin, J. Zheng (2017) Glomerular barrier behaves as an atomically precise bandpass filter in a sub-nanometre regime, Nat. Nanotechnol. 12(11):1096-1102. https://doi.org/10.1038/nnano.2017.170 C.Q. Peng, Y.Y. Huang, J. Zheng (2020) Renal clearable nanocarriers: Overcoming the physiological barriers for precise drug delivery and clearance, J. Control. Release 322:64-80. https://doi.org/10.1016/j.jconrel.2020.03.020 I. Wessels, H.J. Fischer, L. Rink (2021) Dietary and Physiological Effects of Zinc on the Immune System, in: P.J. Stover, R. Balling (Eds.), Annu. Rev. Nutr. 41(1):133-175. https://doi.org/10.1146/annurev-nutr-122019-120635 A.U.R. Khan, K. Huang, M.S. Khalaji, F. Yu, X. Xie, T. Zhu, Y. Morsi, Z. Jinzhong, X. Mo (2021) Multifunctional bioactive core-shell electrospun membrane capable to terminate inflammatory cycle and promote angiogenesis in diabetic wound, Bioact. Mater. 6(9):2783-2800. https://doi.org/10.1016/j.bioactmat.2021.01.040 X. Yang, M. Jia, Z. Li, Z. Ma, J. Lv, D. Jia, D. He, R. Zeng, G. Luo, Y. Yu (2022) In-situ synthesis silver nanoparticles in chitosan/Bletilla striata polysaccharide composited microneedles for infected and susceptible wound healing, Int. J. of Biol. Macromol. 215:550-559. https://doi.org/10.1016/j.ijbiomac.2022.06.131 M.N. Liu, L. Zhang, R.R. Yang, H.Y. Cui, Y.A. Li, X.J. Li, H. Huang (2024) Integrating metal-organic framework ZIF-8 with green modifier empowered bacteria with improved bioremediation, J. Hazard. Mater. 461:132475. https://doi.org/10.1016/j.jhazmat.2023.132475 H. Akhtar, U. Amara, K. Mahmood, M. Hanif, M. Khalid, S. Qadir, Q.H. Peng, M. Safdar, M. Amjad, M.Z. Saif, A. Tahir, M. Yaqub, K. Khalid (2024) Drug carrier wonders: Synthetic strategies of zeolitic imidazolates frameworks (ZIFs) and their applications in drug delivery and anti-cancer activity, Adv. Colloid Interface Sci. 329:103184. https://doi.org/10.1016/j.cis.2024.103184 S. Yang, Y. Zhu, C. Ji, H. Zhu, A. Lao, R. Zhao, Y. Hu, Y. Zhou, J. Zhou, K. Lin, Y. Xu (2024) A five-in-one novel MOF-modified injectable hydrogel with thermo-sensitive and adhesive properties for promoting alveolar bone repair in periodontitis: Antibacterial, hemostasis, immune reprogramming, pro-osteo-/angiogenesis and recruitment, Bioact. Mater. 41:239-256. https://doi.org/10.1016/j.bioactmat.2024.07.016 H. Zhang, S. Yuan, B. Zheng, P. Wu, X. He, Y. Zhao, Z. Zhong, X. Zhang, J. Guan, H. Wang, L. Yang, X. Zheng (2025) Lubricating and Dual-Responsive Injectable Hydrogels Formulated From ZIF-8 Facilitate Osteoarthritis Treatment by Remodeling the Microenvironment, Small 21(3):2407885. https://doi.org/10.1002/smll.202407885 F. Guan, R.X. Wang, Z.J. Yi, P. Luo, W.Y. Liu, Y. Xie, Z.Q. Liu, Z.W. Xia, H. Zhang, Q. Cheng (2025) Tissue macrophages: origin, heterogenity, biological functions, diseases and therapeutic targets, Signal Transduct. Target. Ther. 10(1):93. https://doi.org/10.1038/s41392-025-02124-y N. Eslahi, F. Soleimani, R. Lotfi, F. Mohandes, A. Simchi, M. Razavi (2024) How biomimetic nanofibers advance the realm of cutaneous wound management: The state-of-the-art and future prospects, Prog. Mater. Sci. 145:101293. https://doi.org/10.1016/j.pmatsci.2024.101293 W.L. Liang, J.L. Cheng, J.D. Zhang, Q.Y. Xiong, M.J. Jin, J.H. Zhao (2022) pH-Responsive On-Demand Alkaloids Release from Core-Shell ZnO@ZIF-8 Nanosphere for Synergistic Control of Bacterial Wilt Disease, ACS Nano 16(2):2762-2773. https://doi.org/10.1021/acsnano.1c09724 M.H. Ali, M.A.K. Azad, K.A. Khan, M.O. Rahman, U. Chakma, A. Kumer (2023) Analysis of Crystallographic Structures and Properties of Silver Nanoparticles Synthesized Using PKL Extract and Nanoscale Characterization Techniques, ACS Omega 8(31):28133-28142. https://doi.org/10.1021/acsomega.3c01261 S. Sun, X. Meng, J. Xu, Z. Yang, X. Zhang, H. Dong (2024) Ag-decorated multivariate metal-organic frameworks for enhanced sonodynamic therapy of bacterial infections in wound healing, Nano Today 55:102181. https://doi.org/10.1016/j.nantod.2024.102181 Z. Li, J. Li (2023) Synergistic surface modulation of Ag/ZnO@ZIF-8 hybrid microspheres for enhanced photocatalytic degradation efficiency of Rhodamine B, CrystEngComm 25(14):2064-2074. https://doi.org/10.1039/D2CE01700H L.-X. Liang, J.-C. Li, J.-W. Su, W.-W. Yang, Q. Xu, Z.-G. Qu (2025) ZIF-modified tailored multiscale-pore N-doped graphite felt for high performance vanadium redox flow batteries, J. Energy Storage 128:117252. https://doi.org/10.1016/j.est.2025.117252 J. Cui, D. Wu, Z. Li, G. Zhao, J. Wang, L. Wang, B. Niu (2021) Mesoporous Ag/ZnO hybrid cages derived from ZIF-8 for enhanced photocatalytic and antibacterial activities, Ceram. Int. 47(11):15759-15770. https://doi.org/10.1016/j.ceramint.2021.02.148 N.J. Waleng, T.S. Munonde, A. Mpupa, T. Moremedi, Y. Zhang, P.N. Nomngongo (2025) Photoelectrochemical properties of magnetic amine-based MIL-101(Cr) hybrid material and its application in the degradation of acebutolol in water, RSC Adv. 15(23):17986-17999. https://doi.org/10.1039/D5RA02767E Y.T. Wen, N. Qi, Y. Zhang, Z.Q. Li, B. Fan, M. Zhang, Y. Li (2025)Green synthesis of polysaccharide-based Ag@AgCl photoactive porous hydrogel spheres for enhanced visible light photocatalytic degradation of organic pollutants, Carbohydr. Polym. 356:123401. https://doi.org/10.1016/j.carbpol.2025.123401 S.S. Hu, M.L. Gao, J.J. Huang, H. Wang, Q.Y. Wang, W.J. Yang, Z.H. Sun, X.S. Zheng, H.L. Jiang (2024) Introducing Hydrogen-Bonding Microenvironment in Close Proximity to Single-Atom Sites for Boosting Photocatalytic Hydrogen Production, J. Am. Chem. Soc. 146(29):20391-20400. https://doi.org/10.1021/jacs.4c06013 H. Yuan, H. Shimotani, J. Ye, S. Yoon, H. Aliah, A. Tsukazaki, M. Kawasaki, Y. Iwasa (2010)Electrostatic and electrochemical nature of liquid-gated electric-double-layer transistors based on oxide semiconductors, J. Am. Chem. Soc. 132(51):18402-18407. https://doi.org/10.1021/ja108912x X. Huang, X. Li, C. Deng, X. Deng, Y. Qu, W. Yu, S. Wang, H. Du, S. Zhang, P. Zhang, X. Yue, W. Yu (2025) Fabrication of highly efficient ZnO-Pt catalysts assisted by biomass-derived carboxymethyl cellulose for the photodegradation of diverse antibiotics, J. Environ. Manage. 382:125418. https://doi.org/10.1016/j.jenvman.2025.125418 C.Y. Jiang, M.M. Sun, Y.S. Wang, C.X. Dong, Y. Yu, G. Wang, Y. Lu, Z.B. Chen (2025)Coordination Engineering in Fe-Mn Dual-Atom Nanozyme: Yielding ROS Storm to Efficiently Promote Wound Healing, Adv. Funct. Mater. 2424599. https://doi.org/10.1002/adfm.202424599 Y. Hao, Y. Zhao, C. Zhang, X. Chen, N. Sun, W. Wei (2024) ZIF-8 derived Ag/ZnO photocatalyst with enriched Ag-ZnO interface for effective oxidation of methane to liquid oxygenates with simultaneous mmol-scale productivity and ∼100 % selectivity, Appl. Catal. B Environ. 359:124505. https://doi.org/10.1016/j.apcatb.2024.124505 Y. Zhao, S. Zhao, Y. Du, Z. Gao, Y. Li, H. Ma, H. Li, X. Ren, Q. Fan, D. Wu, Q. Wei (2025)Inverse Oxide/Alloy-Structured Nanozymes with NIR-Triggered Enzymatic Cascade Regulation of ROS Homeostasis for Efficient Wound Healing, Adv. Mater. 37(14):2418731. https://doi.org/10.1002/adma.202418731 P. Hu, Z. Jia, S. Zhao, K. Lin, G. Yang, W. Guo, S. Yu, J. Cheng, G. Du, J. Shi (2024) Injectable Therapeutic Hydrogel with H 2 O 2 Self-Supplying and GSH Consumption for Synergistic Chemodynamic/Low-Temperature Photothermal Inhibition of Postoperative Tumor Recurrence and Wound Infection, Adv. Healthc. Mater. 13(26):2401551. https://doi.org/10.1002/adhm.202401551 Y.L. Lin, X.J. He, T. Huang, J. Zhao, L.Y. Liu, J.Q. He, J.L. Shen, Q.Z. Ren (2024) High-Valent Silver-Porphyrin complex hybrid graphene oxide nanoplatform promotes MRSA-Infected wound healing, Chem. Eng. J. 483:149279. https://doi.org/10.1016/j.cej.2024.149279 T. Nakazono, A.R. Parent, K. Sakai (2015) Improving singlet oxygen resistance during photochemical water oxidation by cobalt porphyrin catalysts, Chemistry 21(18):6723-6726. https://doi.org/10.1002/chem.201500716 L. Xiong, H. Qi, S. Zhang, L. Zhang, X. Liu, A. Wang, J. Tang (2023) Highly Selective Transformation of Biomass Derivatives to Valuable Chemicals by Single-Atom Photocatalyst Ni/TiO 2 , Adv. Mater. 35(16):2209646. https://doi.org/10.1002/adma.202209646 K.L. Wu, D.D. Zhu, X.L. Dai, W.N. Wang, X.Y. Zhong, Z.B. Fang, C. Peng, X.W. Wei, H.S. Qian, X.L. Chen, X.W. Wang, Z.B. Zha, L. Cheng (2022) Bimetallic oxide Cu 1.5 Mn 1.5 O 4 cage-like frame nanospheres with triple enzyme-like activities for bacterial-infected wound therapy, Nano Today 43:101380. https://doi.org/10.1016/j.nantod.2022.101380 R. Łyżeń, G. Gawron, L. Kadziński, B. Banecki (2022) GSH Protects the Escherichia coli Cells from High Concentrations of Thymoquinone, Molecules 27(8):2546. https://doi.org/10.3390/molecules27082546 Y.L. Zhao, Y. Wu, Q. Xu, Y. Liu, Z.Y. Song, H.Y. Han (2024) H 2 O 2 self-supplying and GSH-depleting nanosystem for amplified NIR mediated-chemodynamic therapy of MRSA biofilm-associated infections, J. Nanobiotechnol. 22(1):117. https://doi.org/10.1186/s12951-024-02350-6 Y.L. Fan, H.J. Liu, Z.L. Wang, Z.L. Wang, L.L. Zhu, Y.Z. Wang, F. Song (2024) A One-Nano MOF-Two-Functions Strategy Toward Self-healing, Anti-inflammatory, and Antibacterial Hydrogels for Infected Wound Repair, Chem. Eng. J. 497:155037. https://doi.org/10.1016/j.cej.2024.155037 H. Wang, M. Wang, X. Xu, P. Gao, Z. Xu, Q. Zhang, H. Li, A. Yan, R.Y. Kao, H. Sun (2021)Multi-target mode of action of silver against Staphylococcus aureus endows it with capability to combat antibiotic resistance, Nat. Commun. 12(1):3331. https://doi.org/10.1038/s41467-021-23659-y S.B. Guo, X. Liu, H.H. Chen, J.W. Wang, Y.M. Qiao, T.L. Zhang, X.H. Ji, H. Han, Z.F. Liu, Y. Bai, J. Tang (2024) Antibacterial effect of the metal nanocomposite on Escherichia coli, J. Hazard. Mater. 476:135149. https://doi.org/10.1016/j.jhazmat.2024.135149 E.M. Darby, E. Trampari, P. Siasat, M.S. Gaya, I. Alav, M.A. Webber, J.M.A. Blair (2023) Molecular mechanisms of antibiotic resistance revisited, Nat. Rev. Microbiol. 21(5):280-295. https://doi.org/10.1038/s41579-022-00820-y E.Z. Zhou, Y. Yang, J.L. Li, G.Y. Ren, L.K. Li, Z. Li, Y.P. Lu, T.Y. Gu, F.H. Wang, D.K. Xu (2025) A Novel Cu and Ag Bearing High-Entropy Alloy With Remarkable Antibacterial, Antiviral and Anti-Microbial Corrosion Performances, Adv. Funct. Mater. 35(15):2419467. https://doi.org/10.1016/j.jmst.2020.01.039 L. Tang, S. Xie, D. Wang, Y. Wei, X. Ji, Y. Wang, N. Zhao, Z. Mou, B. Li, W.R. Sun, P.Y. Wang, N.P. Basmadji, J.L. Pedraz, C. Vairo, E.G. Lafuente, M. Ramalingam, X. Xiao, R. Wang (2025) Astragalus polysaccharide/carboxymethyl chitosan/sodium alginate based electroconductive hydrogels for diabetic wound healing and muscle function assessment, Carbohydr. Polym. 350:123058. https://doi.org/10.1016/j.carbpol.2024.123058 L. Pang, C. Dai, L. Bi, Z. Guo, J. Fan (2017) Biosafety and Antibacterial Ability of Graphene and Graphene Oxide In Vitro and In Vivo, Nanoscale Res. Lett. 12(1):564. https://doi.org/10.1186/s11671-017-2317-0 Y. Zhang, X.B. Wei, Y. Xu, W.D. Xia, C.Y. Zheng, H.Y. Zhang, W.T. Chen, K. Xu, Q. Huang (2025) Zinc sulfate gel reshapes the wound microenvironment to promote full-thickness wound healing in mice, Regen. Ther. 28:582-590. https://doi.org/10.1016/j.reth.2025.01.022 M.J. Li, J. Tian, K.K. Yu, H. Liu, X.Q. Yu, N. Wang, Q.M. Gong, K. Li, Y. Shen, X. Wei (2024) A ROS-responsive hydrogel incorporated with dental follicle stem cell-derived small extracellular vesicles promotes dental pulp repair by ameliorating oxidative stress, Bioact. Mater. 36:524-540. https://doi.org/10.1016/j.bioactmat.2024.06.036 W.J. Xiu, L. Wan, K.L. Yang, X. Li, L.H. Yuwen, H. Dong, Y.B. Mou, D.L. Yang, L.H. Wang (2022) Potentiating hypoxic microenvironment for antibiotic activation by photodynamic therapy to combat bacterial biofilm infections, Nat. Commun. 13(1):3875. https://doi.org/10.1038/s41467-022-31479-x F.J. Shariatzadeh, S. Currie, S. Logsetty, R. Spiwak, S. Liu (2025) Enhancing wound healing and minimizing scarring: A comprehensive review of nanofiber technology in wound dressings, Prog. Mater. Sci. 147:101350. https://doi.org/10.1016/j.pmatsci.2024.101350 L. Zhang, Y.F. Lin, L. Hu, Y.N. Wang, C.H. Hu, X.Y. Shangguan, S.Z. Tang, J.C. Chen, P. Hu, Z.S. Chen, Z.F. Ke, Z. Chen (2025)Transient intracellular expression of PD-L1 and VEGFR2 bispecific nanobody in cancer cells inspires long-term T cell activation and infiltration to combat tumor and inhibit cancer metastasis, Mol. Cancer 24(1):1-28. https://doi.org/10.1186/s12943-025-02253-6 Additional Declarations No competing interests reported. Supplementary Files SupportingInformation.docx Cite Share Download PDF Status: Posted Version 1 posted 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. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. 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Technology","correspondingAuthor":false,"prefix":"","firstName":"Xiaohui","middleName":"","lastName":"Ji","suffix":""},{"id":589888607,"identity":"e09d00b7-ebd3-4b3d-9f5f-8709662be261","order_by":11,"name":"Hao Han","email":"","orcid":"","institution":"Shaanxi University of Technology","correspondingAuthor":false,"prefix":"","firstName":"Hao","middleName":"","lastName":"Han","suffix":""},{"id":589888609,"identity":"8a5c6cf9-5486-4902-85d2-489f6807851c","order_by":12,"name":"Shaobo Guo","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA30lEQVRIiWNgGAWjYJACZiBOYGBvgHIPEK2F5wCYIkWLRAKRWuQjkg9/LqixyeOXfP7sMe8PBjm+GwmMnwvwaDG8kZYmPeNYWrHk7BxzY54EBmPJGwnM0jPwaZmRY8bMw3Y4ccPtHDZpoJbEDTcS2Jh58Gsx/szzD6jl5vFnIC31BLXIS+QYSPO2AbXcYDADaUkwIKTFgOdZmjRvH9AvPTlmknPSJAxnnnnYLI3XlnZgiPF8A4YY+/FnEm9sbOT5jicf/IzXlgOofAkgZmzAowFoC37pUTAKRsEoGAVAAAChpkfBhHAEKgAAAABJRU5ErkJggg==","orcid":"","institution":"Shaanxi University of Technology","correspondingAuthor":true,"prefix":"","firstName":"Shaobo","middleName":"","lastName":"Guo","suffix":""}],"badges":[],"createdAt":"2025-10-27 15:42:07","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-7961947/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-7961947/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":102746841,"identity":"d54123b2-fede-4848-9b9d-ad8c28b7d75b","added_by":"auto","created_at":"2026-02-16 09:02:01","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":11780440,"visible":true,"origin":"","legend":"\u003cp\u003eZnO@ZIF-8/Ag composite material for infectious wound healing. (\u003cstrong\u003eA\u003c/strong\u003e) Preparation of ZnO@ZIF-8/Ag composite material. (\u003cstrong\u003eB\u003c/strong\u003e) ZnO@ZIF-8/Ag composite material accelerates wound healing through antibacterial, immunomodulatory, and pro-angiogenic effects\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-7961947/v1/cef94bf580d8a5ca10e3cdbc.png"},{"id":102522220,"identity":"a8e3661a-4f75-4cd7-946a-d78729d2dcc9","added_by":"auto","created_at":"2026-02-12 14:48:56","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":19101167,"visible":true,"origin":"","legend":"\u003cp\u003eCharacterization of the ZnO@ZIF-8/Ag composite materials. TEM images of the ZnO (\u003cstrong\u003eA\u003c/strong\u003e), ZnO@ZIF-8 (\u003cstrong\u003eB\u003c/strong\u003e), and ZnO@ZIF-8/Ag (\u003cstrong\u003eC\u003c/strong\u003e) NPs. (\u003cstrong\u003eD\u003c/strong\u003e) Line scan image of the ZnO@ZIF-8/Ag NPs. (\u003cstrong\u003eE\u003c/strong\u003e) Mapping image of the ZnO@ZIF-8/Ag NPs. XRD patterns (\u003cstrong\u003eF\u003c/strong\u003e), IR spectra (\u003cstrong\u003eG\u003c/strong\u003e), and UV-vis spectra (\u003cstrong\u003eH\u003c/strong\u003e) of the ZnO, ZnO@ZIF-8, and ZnO@ZIF-8/Ag NPs. (\u003cstrong\u003eI\u003c/strong\u003e) XPS spectrum of the ZnO@ZIF-8/Ag NPs; High-resolution XPS spectra of the C 1s (\u003cstrong\u003eJ\u003c/strong\u003e), N 1s (\u003cstrong\u003eK\u003c/strong\u003e), O 1s (\u003cstrong\u003eL\u003c/strong\u003e), Zn 2p (\u003cstrong\u003eM\u003c/strong\u003e), and Ag 3d (\u003cstrong\u003eN\u003c/strong\u003e) peaks in the ZnO@ZIF-8/Ag NPs.\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-7961947/v1/5c5ba00da8b6cc631f4f9101.png"},{"id":102522217,"identity":"2d966624-fdb1-43f8-877a-ad14132067f4","added_by":"auto","created_at":"2026-02-12 14:48:55","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":12829423,"visible":true,"origin":"","legend":"\u003cp\u003eMechanism of ROS generation by the ZnO@ZIF-8/Ag composite materials. (\u003cstrong\u003eA\u003c/strong\u003e) Schematic diagram of the ZnO@ZIF-8/Ag oxidation process. (\u003cstrong\u003eB\u003c/strong\u003e) EIS plots of the ZnO, ZnO@ZIF-8, and ZnO@ZIF-8/Ag NPs. (\u003cstrong\u003eC\u003c/strong\u003e) BE step diagram of Ag\u003csup\u003e+\u003c/sup\u003e adsorption of H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e and O\u003csub\u003e2\u003c/sub\u003e. (\u003cstrong\u003eD\u003c/strong\u003e) Schematic diagram illustrating the mechanism of active oxygen generation by the ZnO@ZIF-8/Ag NPs. UV-vis spectroscopy detection of the self-supply of H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e (\u003cstrong\u003eE\u003c/strong\u003e), ·OH (\u003cstrong\u003eF\u003c/strong\u003e), and \u003csup\u003e1\u003c/sup\u003eO\u003csub\u003e2\u003c/sub\u003e (\u003cstrong\u003eG\u003c/strong\u003e) by the ZnO@ZIF-8/Ag NPs. EPR spectra of the ZnO, ZnO@ZIF-8, and ZnO@ZIF-8/Ag NPs confirming the presence of ·OH (\u003cstrong\u003eH\u003c/strong\u003e) and \u003csup\u003e1\u003c/sup\u003eO\u003csub\u003e2\u003c/sub\u003e (\u003cstrong\u003eI\u003c/strong\u003e). (\u003cstrong\u003eJ\u003c/strong\u003e) Consumption of GSH by the ZnO@ZIF-8/Ag NPs.\u003c/p\u003e","description":"","filename":"3.png","url":"https://assets-eu.researchsquare.com/files/rs-7961947/v1/2789e3ac10d729916a57a1f5.png"},{"id":102522225,"identity":"31c27bfd-4d64-41f3-847c-4090b249a9d9","added_by":"auto","created_at":"2026-02-12 14:48:57","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":33667032,"visible":true,"origin":"","legend":"\u003cp\u003eAntimicrobial activity of ZnO@ZIF-8/Ag. (\u003cstrong\u003eA\u003c/strong\u003e) Schematic diagram of the antibacterial mechanism of ZnO@ZIF-8/Ag. Inhibition zone diameter curves of the ZnO@ZIF-8/Ag NPs against \u003cem\u003eE. coli\u003c/em\u003e (\u003cstrong\u003eB\u003c/strong\u003e), \u003cem\u003eS. aureus \u003c/em\u003e(\u003cstrong\u003eC\u003c/strong\u003e), and DR-\u003cem\u003eSalmonella\u003c/em\u003e (\u003cstrong\u003eD\u003c/strong\u003e). Colony count results of the ZnO@ZIF-8/Ag NPs acting on \u003cem\u003eE. coli\u003c/em\u003e (\u003cstrong\u003eE\u003c/strong\u003e), \u003cem\u003eS. aureus \u003c/em\u003e(\u003cstrong\u003eF\u003c/strong\u003e), and DR-\u003cem\u003eSalmonella\u003c/em\u003e (\u003cstrong\u003eG\u003c/strong\u003e). Colony counts (\u003cstrong\u003eH\u003c/strong\u003e) and antibacterial activity curves (\u003cstrong\u003eI\u003c/strong\u003e) of the ZnO@ZIF-8/Ag NPs against \u003cem\u003eE. coli\u003c/em\u003e, \u003cem\u003eS. aureus\u003c/em\u003e, and DR-\u003cem\u003eSalmonella\u003c/em\u003e. (\u003cstrong\u003eJ\u003c/strong\u003e) Optical images of biofilms stained with crystal violet, n = 3. Data are expressed as mean ± SD; (\u003cstrong\u003eK\u003c/strong\u003e) SEM images and corresponding live/dead bacterial staining assays of untreated \u003cem\u003eE. coli\u003c/em\u003e, \u003cem\u003eS. aureus\u003c/em\u003e, and DR-\u003cem\u003eSalmonella\u003c/em\u003e (control group) and SEM images and live/dead bacterial staining assays of bacteria treated with ZnO@ZIF-8/Ag (experimental group). The live bacteria stained with SYTO-9 exhibit green fluorescence, while the dead bacteria stained with PI exhibit red fluorescence.\u003c/p\u003e","description":"","filename":"4.png","url":"https://assets-eu.researchsquare.com/files/rs-7961947/v1/0f852cc7a014fd82da5c29b3.png"},{"id":102522218,"identity":"7155b4b6-e062-4353-8661-3cfc50ca8034","added_by":"auto","created_at":"2026-02-12 14:48:55","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":58355702,"visible":true,"origin":"","legend":"\u003cp\u003eSchematic diagram of the antibacterial mechanisms of ZnO@ZIF-8/Ag. (\u003cstrong\u003eA\u003c/strong\u003e) Schematic diagram of the interactions between ZnO@ZIF-8/Ag and the \u003cem\u003eE. coli\u003c/em\u003e, \u003cem\u003eS. aureus\u003c/em\u003e, and DR-\u003cem\u003eSalmonella\u003c/em\u003e. (\u003cstrong\u003eB\u003c/strong\u003e) Zeta potential measurements of ZnO@ZIF-8/Ag after interacting with \u003cem\u003eE. coli\u003c/em\u003e, \u003cem\u003eS. aureus\u003c/em\u003e, and DR-\u003cem\u003eSalmonella\u003c/em\u003e for 5 min and 40 min. (\u003cstrong\u003eC\u003c/strong\u003e) Results of the nucleic acid leakage measurements for the three test bacteria after treatment with ZnO@ZIF-8/Ag. (\u003cstrong\u003eD\u003c/strong\u003e) Results of the ion leakage test. (\u003cstrong\u003eE\u003c/strong\u003e) Microcalorimetric detection of cellular respiration. (\u003cstrong\u003eF\u003c/strong\u003e) Molecular docking of Zn\u003csup\u003e2+\u003c/sup\u003e, ZIF-8, and Ag\u003csup\u003e+\u003c/sup\u003e with the protein in \u003cem\u003eβ\u003c/em\u003e-lactamase (PDB code: 4KZ6). (\u003cstrong\u003eG\u003c/strong\u003e) Multi-target antibacterial mechanism of the composite material. Values are expressed as mean ± SD, n = 3.\u003c/p\u003e","description":"","filename":"5.png","url":"https://assets-eu.researchsquare.com/files/rs-7961947/v1/07a8b8ae8d613bbbbf186906.png"},{"id":102522222,"identity":"f9197864-3558-46dd-8b6f-d351cd8f9b7e","added_by":"auto","created_at":"2026-02-12 14:48:57","extension":"png","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":63171077,"visible":true,"origin":"","legend":"\u003cp\u003eBiocompatibility of ZnO@ZIF-8/Ag. (\u003cstrong\u003eA\u003c/strong\u003e) Co-culture of ZnO@ZIF-8/Ag with RAW 264.7 macrophages. (\u003cstrong\u003eB\u003c/strong\u003e) Hemolysis rate of red blood cells after treatment with ZnO@ZIF-8/Ag NPs at different concentration gradients. Insert: Photograph of the hemolysis assay. (\u003cstrong\u003eC\u003c/strong\u003e) Quantitative analysis of cell migration in the scratch assay. (\u003cstrong\u003eD\u003c/strong\u003e) Scratch assay performed on the treated cells. (\u003cstrong\u003eE\u003c/strong\u003e) H\u0026amp;E staining of rat heart, liver, spleen, lung, kidney, and medial gastrocnemius muscle tissue. Values are expressed as mean ± SD, n = 3.\u003c/p\u003e","description":"","filename":"6.png","url":"https://assets-eu.researchsquare.com/files/rs-7961947/v1/f8fbb6fef2ce0fc96c88c893.png"},{"id":102522216,"identity":"e737176e-4a4b-4fab-bfaa-f93030b5e1ab","added_by":"auto","created_at":"2026-02-12 14:48:54","extension":"png","order_by":7,"title":"Figure 7","display":"","copyAsset":false,"role":"figure","size":73874612,"visible":true,"origin":"","legend":"\u003cp\u003eIn vivo antibacterial performance of ZnO@ZIF-8/Ag. (\u003cstrong\u003eA\u003c/strong\u003e) Schematic diagram of the experimental process when treating rats infected with \u003cem\u003eS. aureus\u003c/em\u003e. (\u003cstrong\u003eB\u003c/strong\u003e) Photographs of wounds and simulated wound area diagrams of rats in different treatment groups on days 0, 3, 6, and 9. (\u003cstrong\u003eC\u003c/strong\u003e) Photographs of bacterial colonies in the wound tissue on day 9. (\u003cstrong\u003eD\u003c/strong\u003e) Relative infected area of the wound at days 0, 3, 7, and 9 postsurgery. (\u003cstrong\u003eE\u003c/strong\u003e) Changes in the rat body volume measured from days 0 to 10 postsurgery. (\u003cstrong\u003eF\u003c/strong\u003e) H\u0026amp;E and Masson staining images of the wound tissue from rats at days 3 and 7 posttreatment under different treatment regimens. Values are expressed as mean ± SD, n = 3.\u003c/p\u003e","description":"","filename":"7.png","url":"https://assets-eu.researchsquare.com/files/rs-7961947/v1/d23d85b3119e3a3fbf3ab8f4.png"},{"id":102522223,"identity":"7300d94a-bc0f-48ed-a716-15208cfa1f19","added_by":"auto","created_at":"2026-02-12 14:48:57","extension":"png","order_by":8,"title":"Figure 8","display":"","copyAsset":false,"role":"figure","size":78697312,"visible":true,"origin":"","legend":"\u003cp\u003eFluorescent immunoassay of wound healing with ZnO@ZIF-8/Ag. Immunofluorescence images of representative positive cells for (\u003cstrong\u003eA\u003c/strong\u003e) CD86, (\u003cstrong\u003eB\u003c/strong\u003e) IL-6, (\u003cstrong\u003eC\u003c/strong\u003e) TNF-α, (\u003cstrong\u003eD\u003c/strong\u003e) CD206, (\u003cstrong\u003eE\u003c/strong\u003e) IL-10, (\u003cstrong\u003eF\u003c/strong\u003e) CD31, (\u003cstrong\u003eG\u003c/strong\u003e) VEGF, (\u003cstrong\u003eH\u003c/strong\u003e) COL-Ⅰ, and (\u003cstrong\u003eI\u003c/strong\u003e) FN in wound tissue (n = 3).\u003c/p\u003e","description":"","filename":"8.png","url":"https://assets-eu.researchsquare.com/files/rs-7961947/v1/40ebb4346d3c66f79b7cd6ab.png"},{"id":102522221,"identity":"21d95215-45d4-478c-91a4-4041bf30d988","added_by":"auto","created_at":"2026-02-12 14:48:56","extension":"docx","order_by":0,"title":"","display":"","copyAsset":false,"role":"supplement","size":7296763,"visible":true,"origin":"","legend":"","description":"","filename":"SupportingInformation.docx","url":"https://assets-eu.researchsquare.com/files/rs-7961947/v1/f7ad46674a379cf9ffa3f296.docx"}],"financialInterests":"No competing interests reported.","formattedTitle":"A ZnO@ZIF-8/Ag Dual-Platform Nanocomposite Synergizing ROS Generation for Bactericidal-Immunomodulatory Wound Therapy","fulltext":[{"header":"1 Introduction","content":"\u003cp\u003eThe issue of antibiotic-resistant bacterial infections caused by antibiotic misuse and overuse has become a significant challenge to human health [\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e, \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e]. According to a study published in \u003cem\u003eThe Lancet\u003c/em\u003e, over the course of the next 25 years, the number of fatalities resulting from antibiotic-resistant bacterial infections may exceed 39.1\u0026nbsp;million globally [\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e]. Consequently, an urgent need exists for the development of new antimicrobial strategies to address this growing threat [\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eSilver nanoparticles (Ag NPs) are regarded as a promising alternative to combat the issue of antibiotic resistance owing to their broad-spectrum antimicrobial properties [\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e]. The efficacy of Ag NPs in eliminating pathogens is attributable to multiple bactericidal mechanisms, including the release of metal ions, which disrupt bacterial membrane structures [\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e], the generation of reactive oxygen species (ROS), which interfere with DNA transcription and translation [\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e], and the synergy of metal ions with ROS to inactivate key enzymes [\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e]. This multifaceted approach significantly reduces the risk of antibiotic resistance while ensuring the efficient elimination of pathogenic bacteria. However, the antibacterial activity of Ag NPs is size-dependent, with smaller particles exhibiting stronger antibacterial performance. Notwithstanding, it has been demonstrated that larger specific surface areas increase the likelihood of aggregation and that Ag NPs tend to accumulate in biological organelles, which increases the risk of toxicity [\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e]. Research has shown that when NP sizes are less than 5 nm, they can be excreted safely from the body via the glomerular filtration pores [\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e, \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e]. The development of antimicrobial agents comprising ultra-small Ag NPs that exhibit both good dispersibility and excellent biosafety has thus become a significant challenge for the application of Ag NPs in the medical field.\u003c/p\u003e \u003cp\u003eZinc (Zn) is an essential trace element for the human body. It is involved in the composition and activation of various enzymes within the body and has been shown to play a crucial role in immune system repair [\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e]. Meanwhile, zinc oxide (ZnO), which has antibacterial properties and excellent biocompatibility, serves as an ideal drug carrier in the biomedical field [\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e]. However, when confronted with chronic infection wounds, ZnO appears to demonstrate a limited capacity to effectively inhibit bacterial growth and reproduction [\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e]. Theoretically, the loading of Ag NPs onto the ZnO surface has the potential to address the issue of Ag agglomeration and enhance biocompatibility. However, the presence of negative charges on both Ag and ZnO surfaces has been shown to facilitate the detachment of Ag NPs. Zeolite imidazolium framework material-8 (ZIF-8) is an emerging porous crystalline material. It is widely used as a medium in biomaterials due to its abundant active sites, high biocompatibility, strong antibacterial properties, and cell repair promotion capabilities [\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e]. Furthermore, the imidazole framework contains abundant nitrogen atoms, which possess lone pair electrons capable of forming coordinate bonds with electron-donating atoms or groups (the d electron orbitals of Ag NPs are empty). It can thus be concluded that the 2-methylimidazole (2-MIM) present within ZIF-8 stabilizes the loading of Ag NPs on the surface of ZnO by utilizing coordination bonds and Zn\u003csup\u003e2+\u003c/sup\u003e as active sites. Furthermore, the nitrogen elements within ZIF-8 firmly anchor the Ag NPs [\u003cspan additionalcitationids=\"CR17\" citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eFor the purpose of this study, ZIF-8 was used as an intermediary to synthesize core-shell-type ZnO@ZIF-8/Ag NPs. This was achieved by adsorbing Ag NPs with a particle size of approximately 2.4 nm onto the surface of a ZnO carrier (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003eA). The use of this strategy not only addressed the issue of Ag NP aggregation but also significantly reduced toxicity through the incorporation of ZnO and ZIF-8. The material generated ROS and metal ions in the microenvironment of infected wounds, which resulted in the bacterial cell wall and membrane integrity being compromised and the leakage of bacterial cytoplasm and nucleic acids, thereby ultimately leading to bacterial death. The\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eexperimental results, both in vitro and in vivo, validated the material\u0026rsquo;s high level of biocompatibility, which promoted the polarization of macrophages from M1 to M2 [\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e, \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e] and significantly enhanced wound healing in infected wounds (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003eB). Consequently, this study provides a theoretical foundation for the development of novel anti-infective bioactive materials and offers a novel solution to the issue of antibiotic resistance.\u003c/p\u003e"},{"header":"2 Results and Discussion","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003e2.1 Preparation and Characterization of ZnO@ZIF-8/Ag NPs\u003c/h2\u003e \u003cp\u003eIn this study, ZnO@ZIF-8/Ag NPs were prepared using a layer-by-layer assembly method. Initially, zinc acetate dihydrate (Zn (CH\u003csub\u003e3\u003c/sub\u003eCOO)\u003csub\u003e2\u003c/sub\u003e\u0026middot;2H\u003csub\u003e2\u003c/sub\u003eO) underwent reduction to ZnO NPs through a solvothermal process, in which diethylene glycol (DEG) was used as both the solvent and reducing agent. In the second experiment, Zn\u003csup\u003e2+\u003c/sup\u003e were used as a carrier to coordinate with 2-MIM to achieve the uniform growth of ZIF-8 on its surface. Finally, Ag sol with a particle size of approximately 2.4 nm was adsorbed onto the surface of the ZnO@ZIF-8 to obtain ZnO@ZIF-8/Ag NPs. Transmission electron microscopy (TEM) images revealed the particle sizes of the monodisperse ZnO (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eA), ZnO@ZIF-8 (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eB), and ZnO@ZIF-8/Ag (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eC) as 358.8\u0026thinsp;\u0026plusmn;\u0026thinsp;0.6, 364.6\u0026thinsp;\u0026plusmn;\u0026thinsp;3.3, and 375.7\u0026thinsp;\u0026plusmn;\u0026thinsp;2.8 nm (Fig. \u003cspan refid=\"MOESM1\" class=\"InternalRef\"\u003eS1\u003c/span\u003e), respectively, with the particle size of the ZnO@ZIF-8/Ag/Ag measuring 2.4\u0026thinsp;\u0026plusmn;\u0026thinsp;0.1 nm (Fig. S2). Conversely, the native Ag NPs had a propensity to agglomerate, with an average particle size of 42.4\u0026thinsp;\u0026plusmn;\u0026thinsp;1.5 nm (Fig. S3). Energy-dispersive X-ray spectroscopy (EDS) maps (Fig. S4) and line scans (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eD) demonstrated that the ZnO@ZIF-8/Ag NPs contained C, N, O, Zn, and Ag elements, the respective content of which was 53.42%, 7.11%, 23.95%, 14.15%, and 1.37%. The elemental mapping diagram (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eE) illustrates the uniform distribution of the C (blue), N (yellow), O (red), Zn (green), and Ag (purple) on the ZnO@ZIF-8/Ag surface. X-ray diffraction (XRD) analysis of the material\u0026rsquo;s crystalline structure showed that the ZnO NPs exhibited a wurtzite phase (ICDD 36-1451) [\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e], while the Ag crystalline structure was face-centered cubic (ICDD 04-0783) [\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e] (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eF). The characteristic peaks of ZIF-8 in Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eG, including the C-H stretching vibration (2929 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e), the C\u0026thinsp;=\u0026thinsp;N stretching vibration in the imidazole ring (1583 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e), and the C-N stretching vibrations (1145 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e and 994 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e), indicated that the ZIF-8 successfully coordinated with the ZnO [\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e]. Concurrently, the ultraviolet-visible (UV-vis) molecular absorption spectrum (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eH) of\u003c/p\u003e \u003cp\u003eZnO@ZIF-8 exhibited the characteristic absorption peak of ZIF-8 at 240 nm, while after loading\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003ethe Ag NPs, the local surface plasmon resonance characteristic peaks of the Ag NPs were observed in the wavelength range 400\u0026ndash;450 nm, which indicated a significant red shift and broadening of the spectral band [\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e]. This confirmed the successful loading of the Ag NPs onto the surfaces of the ZnO@ZIF-8 NPs.\u003c/p\u003e \u003cp\u003eX-ray photoelectron spectroscopy (XPS) was utilized to investigate the elemental composition and valence states of the ZnO@ZIF-8/Ag. As demonstrated in Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eI, ZnO@ZIF-8/Ag contained the elements C, N, O, Zn, and Ag. The C 1s spectrum (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eJ) was decomposed into O-C\u0026thinsp;=\u0026thinsp;O (binding energy [BE]\u0026thinsp;=\u0026thinsp;288.0 eV), C-OH (BE\u0026thinsp;=\u0026thinsp;286.3 eV), and C-C (BE\u0026thinsp;=\u0026thinsp;284.8 eV) [\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e]. The peaks at 400.9 eV, 399.8 eV, and 398.7 eV in the N 1s spectrum (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eK) corresponded to N in the imidazole ring [\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e]. The O 1s spectrum (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eL) was fitted within 530.5 and 531.6 eV, which corresponded to the lattice and surface oxygen within the ZnO microspheres. The XPS peaks at 1044.5 and 1021.5 eV were attributed to Zn 2p in the ZnO (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eM) [\u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e]. The XPS spectrum of Ag 3d was assigned to the Ag 3d\u003csub\u003e3/2\u003c/sub\u003e and Ag 3d\u003csub\u003e5/2\u003c/sub\u003e peaks at 373.8 and 367.8 eV (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eN) [\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e].\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec4\" class=\"Section2\"\u003e \u003ch2\u003e2.2 Mechanism of ROS Generation by ZnO@ZIF-8/Ag NPs\u003c/h2\u003e \u003cp\u003eNano-Ag exhibits significant local surface plasmon resonance effects, which induce thermal electron escape during the plasmon resonance process. These high-energy electrons have the capacity to react with oxygen-containing media, and this reaction generates ROS. Furthermore, an established correlation exists between the electron transfer rate and the ROS production rate. Accordingly, under normal conditions, the rate of electron transfer is directly proportional to the level of ROS yield [\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e]. Consequently, electrochemical impedance spectroscopy (EIS) measurements were conducted in this study using a DH7000C electrochemical workstation, and Nyquist plots were generated [\u003cspan additionalcitationids=\"CR29\" citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e]. In a Nyquist plot, a smaller semicircle radius is indicative of\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003elower interfacial resistance and higher electron migration efficiency [\u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e31\u003c/span\u003e]. As shown in the Nyquist plot in Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eB, the radius of ZnO@ZIF-8/Ag is reduced, which suggests enhanced charge transfer efficiency. Density functional theory calculations were then utilized to further investigate the ROS generation process [\u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e32\u003c/span\u003e]. In an oxygen-rich environment, the ZnO@ZIF-8/Ag NPs interacted with the microenvironment of the infected wound to generate H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e, and Ag promoted the breaking of the O-O bonds by providing electrons to produce \u0026middot;OH. Under visible light, surface plasmon resonance occurred on the Ag NP surface, thereby overcoming the potential energy barrier to form \u003csup\u003e1\u003c/sup\u003eO\u003csub\u003e2\u003c/sub\u003e (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eC) [\u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e33\u003c/span\u003e, \u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e34\u003c/span\u003e]. The experimental results demonstrated that Ag can efficiently adsorb H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e (Fig. S5) and catalyze the generation of ROS on the Ag surface. The following equation was postulated:\u003c/p\u003e \u003cp\u003eAg\u0026thinsp;+\u0026thinsp;O\u003csub\u003e2\u003c/sub\u003e \u0026rarr; Ag\u003csup\u003e+\u003c/sup\u003e + \u003csup\u003e1\u003c/sup\u003eO\u003csub\u003e2\u003c/sub\u003e\u003c/p\u003e \u003cp\u003eAg\u0026thinsp;+\u0026thinsp;H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e \u0026rarr; Ag\u003csup\u003e+\u003c/sup\u003e + \u0026middot;OH\u0026thinsp;+\u0026thinsp;OH\u003csup\u003e\u0026ndash;\u003c/sup\u003e\u003c/p\u003e \u003cp\u003eTo investigate the types of ROS generated by the ZnO@ZIF-8/Ag NPs, the following reagents were employed for validation: potassium permanganate (KMnO\u003csub\u003e4\u003c/sub\u003e) [\u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e35\u003c/span\u003e], 3,3',5,5'-tetramethylbenzidine (TMB) [\u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e36\u003c/span\u003e] and 1,3-diphenylisobenzofuran (DPB) [\u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e37\u003c/span\u003e] (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eD). In comparison with the control group (pure potassium permanganate), the ZnO NP, ZnO@ZIF-8 NP, and ZnO@ZIF-8/Ag NP groups all exhibited a decrease in their respective peak absorbances. The most significant decrease was observed in the ZnO@ZIF-8/Ag NP group, which suggested that this group had a superior H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e generation capacity (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eE). Subsequent analysis of the \u0026middot;OH generation capacity (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eF) demonstrated that the ZnO@ZIF-8/Ag NP group exhibited significantly higher levels of absorption compared to the other groups, which confirmed the effective catalysis of \u0026middot;OH generation by this material. Furthermore, the 1,3-diphenylisobenzofuran (DPBF) degradation experiment (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eG) demonstrated that the ZnO@ZIF-8/Ag NP group had the most rapid degradation rate, which indicated that this system generated a greater quantity of \u003csup\u003e1\u003c/sup\u003eO\u003csub\u003e2\u003c/sub\u003e.The results were further validated by electron paramagnetic resonance (EPR) spectroscopy, which confirmed that the 1:2:2:1 quadruplet peak was a characteristic absorption peak of \u0026middot;OH (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eH). In contrast, the single triplet peak at 1:1:1 corresponded to the characteristic peak of \u003csup\u003e1\u003c/sup\u003eO\u003csub\u003e2\u003c/sub\u003e (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eI) [\u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e38\u003c/span\u003e], thus confirming that the ZnO@ZIF-8/Ag NPs possessed excellent synergistic ROS (H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e, \u0026middot;OH, and \u003csup\u003e1\u003c/sup\u003eO\u003csub\u003e2\u003c/sub\u003e).\u003c/p\u003e \u003cp\u003eIn the event of bacterial infections manifesting at local wound sites, macrophages undergo M1-type polarization (pro-inflammatory phenotype) and secrete elevated levels of proinflammatory factors (e.g., TNF-α, IL-6) to facilitate the elimination of pathogens and damaged cells. Concurrently, the body upregulates glutathione (GSH) synthesis to neutralize the oxidative stress and protect the surrounding normal tissues from damage caused by the excessive inflammatory responses [\u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e39\u003c/span\u003e]. However, it has been demonstrated that excessive GSH can be utilized by bacteria to counteract the ROS produced by cells [\u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e40\u003c/span\u003e]. Further, in the context of antimicrobial experiments, the depletion of excessive GSH has been shown to enhance the efficacy of antimicrobials by reducing bacterial tolerance to ROS [\u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e35\u003c/span\u003e, \u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e41\u003c/span\u003e]. As depicted in Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eJ, with an increase in the concentration of ZnO@ZIF-8/Ag NPs, the characteristic absorption peak of GSH at 412 nm declined. It is notable that at a concentration of 50 \u0026micro;g/mL of ZnO@ZIF-8/Ag NPs, the absorption peak underwent a substantial decline. This phenomenon strongly confirms that the ZnO@ZIF-8/Ag NP composite material exhibited significant activity in consuming GSH, which indicates its ability to efficiently catalyze the oxidation of GSH and thereby potentially induce oxidative stress in biological systems.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec5\" class=\"Section2\"\u003e \u003ch2\u003e2.3 In vitro antibacterial activity testing of ZnO@ZIF-8/Ag NPs\u003c/h2\u003e \u003cp\u003eUpon oxygen adsorption, the ZnO@ZIF-8/Ag NPs generated ROS through interactions with wound microenvironment components. Theoretically, these highly oxidative ROS may have effectively oxidized the bacterial cell walls and disrupted the phospholipid bilayers of the cell membranes, which resulted in membrane perforation. Concurrently, the release of Zn\u003csup\u003e2+\u003c/sup\u003e and silver ions (Ag\u003csup\u003e+\u003c/sup\u003e) enabled electrostatic binding to bacterial cell walls, thereby inducing structural alterations, which led to bacterial death [\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e] (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003eA). Three clinically relevant pathogens were selected to evaluate the antibacterial efficacy: gram-negative \u003cem\u003eEscherichia coli\u003c/em\u003e (\u003cem\u003eE. coli\u003c/em\u003e), gram-positive \u003cem\u003eStaphylococcus aureus\u003c/em\u003e (\u003cem\u003eS. aureus\u003c/em\u003e), and drug-resistant \u003cem\u003eSalmonella\u003c/em\u003e (DR-\u003cem\u003eSalmonella\u003c/em\u003e). A disk diffusion assay (Fig. S6) demonstrated that the ZnO and ZnO@ZIF-8 NPs exhibited relatively weak antibacterial activity. In contrast, the ZnO@ZIF-8/Ag NPs displayed broad-spectrum, concentration-dependent antibacterial effects, with significant inhibition against both \u003cem\u003eE. coli\u003c/em\u003e and \u003cem\u003eS. aureus\u003c/em\u003e and notable efficacy against DR-\u003cem\u003eSalmonella\u003c/em\u003e. As illustrated in Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003eB-D, the optimal antibacterial concentration was determined to be 50 \u0026micro;g/mL for the ZnO@ZIF-8/Ag NPs, and this was used in subsequent antibacterial testing. The colony counting results (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003eE-G) confirmed that the composite material achieved 99.9% bactericidal efficiency against all three tested strains within 50 min. Quantitative analysis (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003eH and I) revealed that at the optimal concentration, the ZnO@ZIF-8/Ag NPs exhibited bactericidal rates of 99.9\u0026thinsp;\u0026plusmn;\u0026thinsp;0.2%, 99.7\u0026thinsp;\u0026plusmn;\u0026thinsp;0.12%, and 97.5\u0026thinsp;\u0026plusmn;\u0026thinsp;1.5% against \u003cem\u003eE. coli\u003c/em\u003e, \u003cem\u003eS. aureus\u003c/em\u003e, and DR-\u003cem\u003eSalmonella\u003c/em\u003e, respectively, after 30 min of exposure. As shown in Fig. S7, the minimum inhibitory concentrations (MIC) of the ZnO@ZIF-8/Ag NPs against \u003cem\u003eE. coli\u003c/em\u003e, \u003cem\u003eS. aureus\u003c/em\u003e, and DR-\u003cem\u003eSalmonella\u003c/em\u003e were determined to be 15.63\u0026thinsp;\u0026plusmn;\u0026thinsp;2.3, 31.25\u0026thinsp;\u0026plusmn;\u0026thinsp;0.1, and 31.25\u0026thinsp;\u0026plusmn;\u0026thinsp;0.4 \u0026micro;g/mL, respectively. These results collectively demonstrated that the ZnO@ZIF-8/Ag NPs exhibited outstanding antibacterial performance across the three distinct evaluation methods (disk diffusion assay, colony counting, and MIC\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003edetermination), which highlights the potential of ZnO@ZIF-8/Ag as a highly effective antimicrobial material.\u003c/p\u003e \u003cp\u003eDuring the process of reproduction, bacteria form biofilms in suitable external environments. The formation of biofilms is a multi-stage dynamic process that typically includes the following steps: adhesion, where bacteria reversibly attach to solid surfaces through physical and chemical interactions, and colonization, where microorganisms form irreversible, robust bonds with surfaces through the secretion of extracellular polymers. As microorganisms proliferate and extracellular polymers accumulate further, the biofilm gradually matures to form a complex three-dimensional structure with bacteria inside, which exhibits high antibiotic resistance and metabolic diversity [\u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e36\u003c/span\u003e]. Biofilms provide an effective protective barrier for bacteria by shielding them from phagocytosis and neutralizing antimicrobial agents. This, in turn, safeguards bacteria from host immune defense attacks. To assess the anti-biofilm activity of the ZnO@ZIF-8/Ag NPs, crystal violet staining experiments were conducted using three representative bacterial strains (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003eJ). The control group exhibited biofilms characterized by robust structural integrity, while the biofilm biomass in the treated group was found to be significantly diminished. Quantitative analysis (Fig. S8) confirmed that the ZnO@ZIF-8/Ag NPs significantly inhibited the formation of biofilms in \u003cem\u003eE. coli\u003c/em\u003e and \u003cem\u003eS. aureus\u003c/em\u003e and demonstrated some inhibitory effects on the formation of biofilms in DR-\u003cem\u003eSalmonella\u003c/em\u003e. These findings pointed to the possibility of using ZnO@ZIF-8/Ag NPs as an effective strategy against biofilm-associated infections. The inhibitory effect could be attributed to the following mechanisms: the disruption of bacterial adhesion, interference with EPS production, and the exertion of direct bactericidal effects on the cells embedded within the biofilm [\u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e42\u003c/span\u003e].\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec6\" class=\"Section2\"\u003e \u003ch2\u003e2.4 Research on antibacterial mechanisms\u003c/h2\u003e \u003cp\u003eThe primary components of bacterial cell walls (e.g., phospholipid layers, teichoic acid, lipopolysaccharides) are rich in negatively charged functional groups, such as phosphate (PO\u003csub\u003e4\u003c/sub\u003e\u003csup\u003e3\u0026ndash;\u003c/sup\u003e), carboxyl (COO\u003csup\u003e\u0026minus;\u003c/sup\u003e), and hydroxyl (OH\u003csup\u003e\u0026minus;\u003c/sup\u003e) groups. These structures are imperative for preserving bacterial integrity and metabolic functions. Theoretically, the metal ions released by the ZnO@ZIF-8/Ag NPs may have disrupted the bacterial cell walls through electrostatic adsorption, while the ROS generated by the nanomaterials oxidized the phospholipid molecules, thereby leading to cell wall lysis [\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e, \u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e, \u003cspan citationid=\"CR43\" class=\"CitationRef\"\u003e43\u003c/span\u003e] (Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003eA). To validate this hypothesis, a series of experiments were designed with the aim of assessing the antibacterial mechanisms of the nanocomposites. The ion release experiments (Fig. S9) confirmed that the ZnO@ZIF-8/Ag NPs released 78.5\u0026thinsp;\u0026plusmn;\u0026thinsp;0.4% Zn\u003csup\u003e2+\u003c/sup\u003e and 67.4\u0026thinsp;\u0026plusmn;\u0026thinsp;0.1% Ag\u003csup\u003e+\u003c/sup\u003e. Scanning electron microscopy (SEM) was used to discern the effects of the material on the morphological alterations of the bacteria. In the control group, the bacteria exhibited smooth surfaces with no obvious cytoplasmic leakage. In the experimental group, the bacterial cell surfaces exhibited clear indications of damage, which manifested as wrinkling and denting. Among the tested organisms, the cell membranes of \u003cem\u003eE. coli\u003c/em\u003e and \u003cem\u003eS. aureus\u003c/em\u003e were the most severely compromised, and this damage was accompanied by substantial leakage of cytoplasmic content. Fluorescence inverted microscopy showed that the bacteria treated with ZnO@ZIF-8/Ag NPs exhibited red fluorescence. This finding suggests that the integrity of the cell membrane structure was compromised, which enabled PI to enter the cell interior and bind to nucleic acids. In contrast, the untreated control bacteria primarily exhibited green fluorescence, which indicated intact cell membrane structures and viable bacterial cells (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003eK) [\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e]. These results demonstrated that the ZnO@ZIF-8/Ag NPs exhibited highly effective broad-spectrum antibacterial activity against \u003cem\u003eE. coli\u003c/em\u003e, \u003cem\u003eS. aureus\u003c/em\u003e, and DR-\u003cem\u003eSalmonella\u003c/em\u003e.\u003c/p\u003e \u003cp\u003eIt was hypothesized that alterations in the potential of bacterial cell walls may be indicative of changes in the surface charge of bacterial cell membranes. The Zeta potential values for \u003cem\u003eE. coli\u003c/em\u003e, \u003cem\u003eS. aureus\u003c/em\u003e, and DR-\u003cem\u003eSalmonella\u003c/em\u003e were \u0026minus;\u0026thinsp;15.59\u0026thinsp;\u0026plusmn;\u0026thinsp;1.0, \u0026minus;\u0026thinsp;14.83\u0026thinsp;\u0026plusmn;\u0026thinsp;2.4, and \u0026minus;\u0026thinsp;14.35\u0026thinsp;\u0026plusmn;\u0026thinsp;1.7 mV, respectively. Following a period of 5 min during to allow for their interaction with the ZnO@ZIF-8/Ag NPs, the Zeta potentials of the three bacteria increased to \u0026minus;\u0026thinsp;10.85\u0026thinsp;\u0026plusmn;\u0026thinsp;2.2, \u0026minus;\u0026thinsp;11.91\u0026thinsp;\u0026plusmn;\u0026thinsp;0.1, and \u0026minus;\u0026thinsp;12.64\u0026thinsp;\u0026plusmn;\u0026thinsp;0.3 mV, respectively. After a further 40 min of interaction, these potentials increased further to \u0026minus;\u0026thinsp;4.23\u0026thinsp;\u0026plusmn;\u0026thinsp;1.2, \u0026minus;\u0026thinsp;5.33\u0026thinsp;\u0026plusmn;\u0026thinsp;0.8, and \u0026minus;\u0026thinsp;7.06\u0026thinsp;\u0026plusmn;\u0026thinsp;1.9 mV, respectively (Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003eB). The results obtained from this study indicate that ZnO@ZIF-8/Ag NPs have the capacity to significantly neutralize the negative charges on the surfaces of bacterial cell membranes. Furthermore, the neutralization effect was found to be time-dependent. The data pertaining to the leakage of bacterial nucleic acid and cytoplasm (Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003eC) demonstrated that the nucleic acid leakage in the experimental group increased by 0.58\u0026thinsp;\u0026plusmn;\u0026thinsp;0.1, 0.35\u0026thinsp;\u0026plusmn;\u0026thinsp;2.9, and 0.21\u0026thinsp;\u0026plusmn;\u0026thinsp;1.3 for \u003cem\u003eE. coli\u003c/em\u003e, \u003cem\u003eS. aureus\u003c/em\u003e, and DR-\u003cem\u003eSalmonella\u003c/em\u003e, respectively. The ion leakage experiments (Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003eD) revealed significant reductions in ionic content compared to the control group. In \u003cem\u003eE. coli\u003c/em\u003e, \u003cem\u003eS. aureus\u003c/em\u003e, and DR-\u003cem\u003eSalmonella\u003c/em\u003e, respectively: K⁺ levels decreased by 6.72\u0026thinsp;\u0026plusmn;\u0026thinsp;0.1, 4.71\u0026thinsp;\u0026plusmn;\u0026thinsp;0.9, and 5.88\u0026thinsp;\u0026plusmn;\u0026thinsp;0.1 mg/mL; Ca\u0026sup2;⁺ levels decreased by 8.91\u0026thinsp;\u0026plusmn;\u0026thinsp;0.3, 3.37\u0026thinsp;\u0026plusmn;\u0026thinsp;1.2, and 3.85\u0026thinsp;\u0026plusmn;\u0026thinsp;1.1 mg/mL; Mg\u0026sup2;⁺ levels decreased by 0.23\u0026thinsp;\u0026plusmn;\u0026thinsp;0.1, 0.55\u0026thinsp;\u0026plusmn;\u0026thinsp;0.3, and 0.1\u0026thinsp;\u0026plusmn;\u0026thinsp;0.05 mg/mL. These data indicate that ZnO@ZIF-8/Ag has the capacity to compromise the integrity of bacterial cell membranes, leading to the efflux of intracellular ions and nucleic acids. Furthermore, microcalorimetric analysis (Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003eE) demonstrated the absence of an exponential growth phase in the bacterial growth curve\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003efollowing treatment with ZnO@ZIF-8/Ag, with the majority of the heat output exhibiting stabilization. This suggests that ZnO@ZIF-8/Ag may inhibit bacterial energy metabolism by disrupting the respiratory systems of bacteria, thereby exerting its antibacterial effect [\u003cspan citationid=\"CR44\" class=\"CitationRef\"\u003e44\u003c/span\u003e].\u003c/p\u003e \u003cp\u003e \u003cem\u003eβ\u003c/em\u003e-Lactamases have been identified as significant contributors to bacterial antibiotic resistance, as they facilitate bacterial survival and proliferation in environments exposed to antibiotics. This phenomenon represents a substantial challenge in the field of clinical antimicrobial therapy [\u003cspan citationid=\"CR45\" class=\"CitationRef\"\u003e45\u003c/span\u003e]. To assess the hypothesis that ZnO@ZIF-8/Ag NPs enhance antimicrobial efficacy by inhibiting \u003cem\u003eβ\u003c/em\u003e-lactamase activity, the binding affinities of Zn\u003csup\u003e2+\u003c/sup\u003e, ZIF-8 and Ag\u003csup\u003e+\u003c/sup\u003e with \u003cem\u003eβ\u003c/em\u003e-lactamase were calculated separately (Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003eF). The Bes of Zn\u003csup\u003e2+\u003c/sup\u003e, ZIF-8, and Ag\u003csup\u003e+\u003c/sup\u003e with \u003cem\u003eβ\u003c/em\u003e-lactamase were \u0026minus;\u0026thinsp;0.82, \u0026minus;\u0026thinsp;5.03, and \u0026minus;\u0026thinsp;0.21 kcal/mol, respectively (Fig. S10), which indicated that the inhibitory capacity against \u003cem\u003eβ\u003c/em\u003e-lactamase was ZIF-8\u0026thinsp;\u0026gt;\u0026thinsp;Zn\u003csup\u003e2+\u003c/sup\u003e \u0026gt; Ag\u003csup\u003e+\u003c/sup\u003e. These results demonstrated that the nanocomposite likely exerted synergistic antibacterial effects through multiple mechanisms, as schematically illustrated in Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003eG. Bacterial surfaces typically carry negative charges, so this enabled strong electrostatic interactions between the surface components (e.g., carboxyl and phosphate groups) and the released Zn\u003csup\u003e2+\u003c/sup\u003e/Ag\u003csup\u003e+\u003c/sup\u003e ions from the ZnO@ZIF-8/Ag NPs. Concurrently, the unique physical structure of ZIF-8 induced mechanical damage to the bacterial membranes upon contact, which compromised the membrane integrity. This structural disruption significantly altered the membrane permeability, disrupted the osmotic balance and caused massive leakage of the intracellular components (e.g., proteins, nucleic acids, ions), and thereby severely interfered with the normal bacterial physiology. Furthermore, the ZnO@ZIF-8/Ag NPs exhibited catalytic activity, which facilitated the adsorption and subsequent conversion of environmental H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e and O\u003csub\u003e2\u003c/sub\u003e into highly oxidative hydroxyl radicals (\u0026middot;OH) and singlet oxygen (\u003csup\u003e1\u003c/sup\u003eO\u003csub\u003e2\u003c/sub\u003e). These ROS inflicted oxidative damage on critical biomacromolecules within the bacterial cells. Simultaneously, the nanocomposite oxidized the intracellular reduced GSH to oxidized glutathione (GSSG), which amplified the ROS-mediated antibacterial effects and disrupted the cellular redox homeostasis. The released Ag\u003csup\u003e+\u003c/sup\u003e ions specifically bound to the thiol groups on the bacterial DNA, thereby inducing strand breaks and base damage that compromised the genetic integrity of the bacteria. Additionally, Ag\u003csup\u003e+\u003c/sup\u003e disrupted the ion channels in the bacterial membranes, which led to the leakage of essential ions (e.g., K⁺, Na⁺, Ca\u0026sup2;⁺) and consequent loss of cellular function. In summary, the ZnO@ZIF-8/Ag NPs achieved potent antibacterial efficacy through a multifaceted mechanism involving (1) electrostatic adsorption, (2) physical membrane damage, (3) ROS generation, (4) enzymatic interference, (5) DNA damage, and (6) ion channel disruption [\u003cspan citationid=\"CR46\" class=\"CitationRef\"\u003e46\u003c/span\u003e].\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec7\" class=\"Section2\"\u003e \u003ch2\u003e2.5 Biocompatibility of ZnO@ZIF-8/Ag NPs\u003c/h2\u003e \u003cp\u003eThe biological safety of materials utilized in wound dressings must be a primary consideration during evaluations. In this study, the biocompatibility of the ZnO@ZIF-8/Ag NPs was systematically assessed through a range of in vitro and in vivo methods, including cell co-culture, blood compatibility, cell migration, and tissue toxicity lesions.\u003c/p\u003e \u003cp\u003eIn the experimental setup, RAW 264.7 macrophages were co-cultured with ZnO@ZIF-8/Ag\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eNPs. To distinguish between the macrophages and the ZnO@ZIF-8/Ag NPs, the macrophages were labelled with a fluorescent tag constructed using lentivirus and appeared red. Meanwhile, the ZnO@ZIF-8/Ag NPs were labelled with CY5-polyethylene glycol-sulfhydryl (CY5-PEG-SH) to appear green. Following phagocytosis of the ZnO@ZIF-8/Ag NPs, the cells exhibited yellow fluorescence. The results (Fig.\u0026nbsp;\u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e6\u003c/span\u003eA) demonstrated the effective uptake of the ZnO@ZIF-8/Ag NPs by the macrophages. As depicted in Fig.\u0026nbsp;\u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e6\u003c/span\u003eB, hemocompatibility testing revealed that, within the concentration range that was examined, treatment with ZnO@ZIF-8/Ag NPs did not cause significant hemolysis. The observed hemolysis rates were all found to be below the internationally recognized safety threshold of 5% [\u003cspan citationid=\"CR47\" class=\"CitationRef\"\u003e47\u003c/span\u003e]. Subsequently, scratch wound healing experiments were conducted using L929 mouse fibroblast cells (Fig.\u0026nbsp;\u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e6\u003c/span\u003eC and D). In comparison with the cells devoid of ZnO@ZIF-8/Ag NPs, the cell migration rate in the ZnO@ZIF-8/Ag NP-treated group was significantly increased, reaching 68.9% at 12 h and 92.6% at 24 h. Furthermore, a series of histopathological examinations using hematoxylin and eosin (H\u0026amp;E) staining were conducted on major organs (heart, liver, spleen, lungs, and kidneys) of Sprague-Dawley rats, along with in vivo toxicity assessments (Fig.\u0026nbsp;\u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e6\u003c/span\u003eE). No overt pathological changes were observed in the cellular morphology and histological structure of these organs. The results of the in vivo muscle injection experiment showed that the experimental group exhibited mild local muscle fiber ruptures, with no obvious cellular swelling, degeneration, or necrosis. These alterations were also observed in normal skeletal muscle cells, which indicated that the experimental treatment had minimal interference with basic cellular physiological functions [\u003cspan citationid=\"CR48\" class=\"CitationRef\"\u003e48\u003c/span\u003e]. In summary, the experimental findings from both the in vitro and in vivo contexts demonstrated that the ZnO@ZIF-8/Ag NPs exhibited favorable biocompatibility and promoted cell proliferation.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec8\" class=\"Section2\"\u003e \u003ch2\u003e2.6 Antimicrobial and wound healing properties in vivo\u003c/h2\u003e \u003cp\u003eZinc is a trace element that is indispensable to the human body, as it plays a crucial role in\u003c/p\u003e \u003cp\u003efundamental life processes, such as DNA synthesis, protein metabolism, cell proliferation, and immune regulation. Research has demonstrated that when zinc is applied directly to wounds and the surrounding damaged skin, it promotes the clearance of necrotic tissue, reduces the risk of infection, alleviates inflammation, and activates epithelialization [\u003cspan citationid=\"CR49\" class=\"CitationRef\"\u003e49\u003c/span\u003e]. In this study, a full-thickness skin\u003c/p\u003e \u003cp\u003e \u003cem\u003eS. aureus\u003c/em\u003e infectious wound model was established in Sprague-Dawley rats to evaluate the in vivo therapeutic efficacy of ZnO@ZIF-8/Ag (Fig.\u0026nbsp;\u003cspan refid=\"Fig7\" class=\"InternalRef\"\u003e7\u003c/span\u003eA and B). On the third day, the ZnO@ZIF-8/Ag group exhibited a 49.48\u0026thinsp;\u0026plusmn;\u0026thinsp;3.9% healing rate, which increased to 93.23\u0026thinsp;\u0026plusmn;\u0026thinsp;1.5% on the sixth day and 99.78\u0026thinsp;\u0026plusmn;\u0026thinsp;1.7% on the ninth day when compared with the control, ZnO, and ZnO@ZIF-8 groups (Fig.\u0026nbsp;\u003cspan refid=\"Fig7\" class=\"InternalRef\"\u003e7\u003c/span\u003eD). Furthermore, the wounds treated with ZnO@ZIF-8/Ag exhibited no bacterial presence after day 9 (Fig.\u0026nbsp;\u003cspan refid=\"Fig7\" class=\"InternalRef\"\u003e7\u003c/span\u003eC and Fig. S11). Dynamic monitoring revealed no significant difference in body weight between the \u003cem\u003eS. aureus\u003c/em\u003e-infected group and the control group (Fig.\u0026nbsp;\u003cspan refid=\"Fig7\" class=\"InternalRef\"\u003e7\u003c/span\u003eE), which confirmed the significant advantages and application potential of ZnO@ZIF-8/Ag in the healing of infected wounds.\u003c/p\u003e \u003cp\u003eTo further assess the process of wound healing in infected wounds, wound sections were collected on days 3 and 7, and H\u0026amp;E staining and Masson\u0026rsquo;s trichrome staining were performed. As demonstrated in Fig.\u0026nbsp;\u003cspan refid=\"Fig7\" class=\"InternalRef\"\u003e7\u003c/span\u003eF, during the acute phase post-trauma (day 3), the control group exhibited extensive infiltration of inflammatory cells (primarily neutrophils and macrophages) and a loosened tissue structure; in contrast, the ZnO@ZIF-8/Ag-treated group had significantly reduced inflammatory cell infiltration and better reserved tissue structural integrity. By the repair phase (day 7), residual inflammatory cells and minimal collagen deposition were still visible in the control group, whereas the ZnO@ZIF-8/Ag group exhibited nearly complete resolution of the inflammatory response, with newly formed epithelial tissue intact. The dermis layer had abundant vascularization and orderly collagen fibers, with significantly increased collagen deposition. The results indicated that the ZnO@ZIF-8/Ag NPs material exhibited significant advantages in inhibiting inflammatory responses, promoting epithelial formation, and enhancing collagen deposition.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec9\" class=\"Section2\"\u003e \u003ch2\u003e2.7 Immunofluorescence\u003c/h2\u003e \u003cp\u003eThe process of wound healing is categorized into four distinct stages: hemostasis, inflammation, proliferation, and remodeling [\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e]. During the inflammatory phase, M1 (marked by CD86) initiate the inflammatory response by secreting pro-inflammatory factors, such as IL-6 and TNF-α, to eliminate pathogens and necrotic tissue. M2 (marked by CD206) subsequently promote tissue repair and angiogenesis by secreting anti-inflammatory factors, such as IL-10, and stimulating the body to produce pro-repair factors, such as platelet-endothelial cell adhesion molecule-1 (CD31), vascular endothelial growth factor (VEGF), type I collagen/fibronectin (COL-Ⅰ/FN), and proliferating cell nuclear antigen (PCNA). These factors participate in intercellular interactions and signal transduction and collectively drive the transition of wounds from the inflammatory phase to the proliferative phase, thereby ensuring the smooth progression of the wound healing process [\u003cspan citationid=\"CR50\" class=\"CitationRef\"\u003e50\u003c/span\u003e, \u003cspan citationid=\"CR51\" class=\"CitationRef\"\u003e51\u003c/span\u003e]. In the immunofluorescence images in this study, the distribution of CD86 (Fig.\u0026nbsp;\u003cspan refid=\"Fig8\" class=\"InternalRef\"\u003e8\u003c/span\u003eA), IL-6 (Fig.\u0026nbsp;\u003cspan refid=\"Fig8\" class=\"InternalRef\"\u003e8\u003c/span\u003eB), and TNF-α (Fig.\u0026nbsp;\u003cspan refid=\"Fig8\" class=\"InternalRef\"\u003e8\u003c/span\u003eC) was lower in the ZnO@ZIF-8/Ag group. The expression levels of CD206 (Fig.\u0026nbsp;\u003cspan refid=\"Fig8\" class=\"InternalRef\"\u003e8\u003c/span\u003eD) and IL-10 (Fig.\u0026nbsp;\u003cspan refid=\"Fig8\" class=\"InternalRef\"\u003e8\u003c/span\u003eE) were elevated in the ZnO, ZnO@ZIF-8, and ZnO@ZIF-8/Ag groups compared to the group without added material, but particularly in the ZnO@ZIF-8/Ag group. This\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003efinding indicated that ZnO@ZIF-8/Ag effectively promoted the polarization of M1 macrophages toward M2 macrophages, reduced the destructive effects of inflammatory factors on tissues, enhanced the role of M2 macrophages in tissue repair, and accelerated the wound healing process.\u003c/p\u003e \u003cp\u003eIn the context of angiogenesis, CD31 is a marker for vascular endothelial cells, while VEGF is a key player in the process, with its expression levels reflecting the status of angiogenesis. COL-Ⅰ/FN and PCNA are significant components of the extracellular matrix, they play pivotal roles in cell adhesion, proliferation, and migration and serve as key markers for tissue repair during wound healing [\u003cspan citationid=\"CR52\" class=\"CitationRef\"\u003e52\u003c/span\u003e, \u003cspan citationid=\"CR53\" class=\"CitationRef\"\u003e53\u003c/span\u003e]. In the ZnO@ZIF-8/Ag sample, the expression levels of CD31 (Fig.\u0026nbsp;\u003cspan refid=\"Fig8\" class=\"InternalRef\"\u003e8\u003c/span\u003eF), VEGF (Fig.\u0026nbsp;\u003cspan refid=\"Fig8\" class=\"InternalRef\"\u003e8\u003c/span\u003eG), COL-Ⅰ (Fig.\u0026nbsp;\u003cspan refid=\"Fig8\" class=\"InternalRef\"\u003e8\u003c/span\u003eH), FN (Fig.\u0026nbsp;\u003cspan refid=\"Fig8\" class=\"InternalRef\"\u003e8\u003c/span\u003eI), and PCNA (Fig. S12) were found to be significantly elevated in comparison to those in the ZnO and ZnO@ZIF-8 samples. It is clear from the experimental evidence that ZnO@ZIF-8/Ag significantly advances the inhibition of inflammatory responses, the promotion of tissue repair, the acceleration of angiogenesis, and the enhancement of extracellular matrix synthesis, all of which provides robust support for its application in wound healing.\u003c/p\u003e \u003c/div\u003e"},{"header":"3 Conclusion","content":"\u003cp\u003eThis study involved the use of ZnO as a precursor and employed a ZIF-8 coordination-mediated strategy to synthesize ZnO@ZIF-8/Ag nanocomposites loaded with ultra-small Ag sol (approximately 2.4 nm) with the aim of treating wounds caused by drug-resistant bacteria. These nanocomposites exhibited high antibacterial activity and biocompatibility and promoted the healing of infected wounds. The antibacterial activity of ZnO@ZIF-8/Ag can be attributed to the generation of ROS and the release of Zn\u003csup\u003e2+\u003c/sup\u003e and Ag\u003csup\u003e+\u003c/sup\u003e ions with the simultaneous inactivation of bacteria through the regulation of the redox balance in the microenvironment through the removal of excess GSH. The ZnO@ZIF-8/Ag NPs demonstrated high levels of biological activity, as indicated by both the in vitro and in vivo biocompatibility results. Moreover, this nanocomposite material induced M1 macrophages to polarize toward the M2 phenotype, thereby reducing inflammatory responses in the in vivo experiments, promoting collagen synthesis and deposition, speeding up angiogenesis and tissue remodeling, and ultimately accelerating wound healing rates and achieving comprehensive tissue repair. In the present study, a novel nano-composite material-based synergistic treatment strategy for bacterial infection wounds was developed. This strategy has significant application potential and provides a theoretical basis and experimental foundation for the design of novel antimicrobial wound dressings.\u003c/p\u003e"},{"header":"4 Experimental section","content":"\u003cp\u003eFor all the experiments, please see the Supporting Information.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eSupplementary Information\u003c/strong\u003e The online version contains supplementary material available at\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthor contributions\u0026nbsp;\u003c/strong\u003eRuiling Hu: Writing\u0026ndash;original draft, Software, Methodology, Data curation. Yujie Zhang: Writing\u0026ndash;original draft, Investigation, Formal analysis, Data curation. Chengzhi Gao: Writing\u0026ndash;original draft, Software, Data curation. Hang Wei: Investigation, Data curation. Xianjian Hong: Formal analysis, Validation, Investigation. Tinghui Qiang: Validation, Software, Methodology, Formal analysis. Zhongshang Guo: Formal analysis, Data curation. Li Huang: Methodology, Investigation. Xiaoyun Lei: Software, Methodology, Investigation, Formal analysis. Caibin Zhao: Investigation, Software, Formal analysis, Data curation. Xiaohui Ji: Writing\u0026ndash;review \u0026amp; editing, Methodology, Funding acquisition, Project administration. Hao Han: Writing\u0026ndash;review \u0026amp; editing, Methodology. Shaobo Guo: Writing\u0026ndash;review \u0026amp; editing, Supervision, Investigation, Funding acquisition, Conceptualization.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding\u003c/strong\u003e This work was supported by the Science and Technology Innovation Team Project of Shaanxi Province (2025RS-CXTD-040), Shaanxi Provincial Natural Science Foundation (24JK0376), Key Research and Development Program of Shaanxi (2024SF-YBXM-158) and Shaanxi Province Health and Wellness Scientific Research Innovation Capacity Enhancement Project (2024PT-16). Additional funding was provided by the Open Project Program of the State Key Laboratory of Qinba Biological Resources and Ecological Environment (SLGPT2019KF04-01).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eData availability\u003c/strong\u003e Data will be made available on request.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCompeting interests\u003c/strong\u003e The authors declare no competing interests.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eX. Wang, X.D. Qin, Y. Liu, Y.T. Fang, H. Meng, M.L. Shen, L.L. Liu, W.W. Huan, J. Tian, Y.W. Yang (2024) Plasmonic Supramolecular Nanozyme-Based Bio-Cockleburs for Synergistic Therapy of Infected Diabetic Wounds, Adv. Mater. 36(49):2411194. https://doi.org/10.1002/adma.202411194\u003c/li\u003e\n\u003cli\u003eA. Abbas, A. Barkhouse, D. Hackenberger, G.D. Wright (2024) Antibiotic resistance: A key microbial survival mechanism that threatens public health, Cell Host Microbe 32(6):837-851. https://doi.org/10.1016/j.chom.2024.05.015\u003c/li\u003e\n\u003cli\u003eM. Naghavi, S. E. Vollset, K. S. Ikuta, L. R. Swetschinski, A. P. Gray, E. E. Wool, et al., (2024) Global burden of bacterial antimicrobial resistance 1990\u0026ndash;2021: a systematic analysis with forecasts to 2050, Lancet 404(10459):1199-1226. https://doi.org/10.1016/S0140-6736(24)01867-1\u003c/li\u003e\n\u003cli\u003eJ. Song, Y.H. Lu, T. Pan, J.F. Wang, Z.Q. Liu, L. Xu, S.Q. Zhang, Y.P. Li, Y.Y. Bai, B.C. Heng, X.N. Zheng, H.Y. Wu, X.Y. Han, Y.S. Guo, Q. Cui, X.L. Deng, X.H. Zhang (2024) Manipulation of Surface Electrical Charge on Nanocomposite Membranes Confers Wide Spectrum Bactericidal Effects and Promotes Tissue Regeneration, Adv. Funct. Mater. 34(22):2314024. https://doi.org/10.1002/adfm.202314024\u003c/li\u003e\n\u003cli\u003eX.Y. Gong, N.D. Jadhav, V.V. Lonikar, A.N. Kulkarni, H.K. Zhang, B.R. Sankapal, J.N. Ren, B. Bin Xu, H.M. Pathan, Y. Ma, Z.P. Lin, E. Witherspoon, Z. Wang, Z.H. Guo (2024) An overview of green synthesized silver nanoparticles towards bioactive antibacterial, antimicrobial and antifungal applications, Adv. Colloid Interface Sci. 323:103053. https://doi.org/10.1016/j.cis.2023.103053\u003c/li\u003e\n\u003cli\u003eC.Y. Wang, S.L. Zhu, Y.Q. Liang, C.L. Qin, F. Wang, H. Wang, C.T. Chang (2023) A. Inoue, Flexible free-standing antibacterial nanoporous Ag ribbon, J. Colloid Interface Sci. 645:287-296. https://doi.org/10.1016/j.jcis.2023.04.153\u003c/li\u003e\n\u003cli\u003eY.X. Wang, M.J. Malkmes, C. Jiang, P. Wang, L.Y. Zhu, H.M. Zhang, Y.H. Zhang, H. Huang, L. Jiang (2021) Antibacterial mechanism and transcriptome analysis of ultra-small gold nanoclusters as an alternative of harmful antibiotics against Gram-negative bacteria, J. Hazard. Mater. 416:126236. https://doi.org/10.1016/j.jhazmat.2021.126236\u003c/li\u003e\n\u003cli\u003eD.L. Han, X.M. Liu, S.L. Wu (2022) Metal organic framework-based antibacterial agents and their underlying mechanisms, Chem. Soc. Rev. 51(16):7138-7169. https://doi.org/10.1039/D2CS00460G\u003c/li\u003e\n\u003cli\u003eZ. Li, T. Yang, X. Li, P. Yin, B. Yang, D. Li, Y. Wang, W. Teng, Q. Yu, W. Li (2025) A Bio-Responsive Hydrogel with Spatially Heterogeneous Structure for Treating Infectious Tissue Injuries, Adv Sci (Weinh) 12(23):2500088. https://doi.org/10.1002/advs.202570182\u003c/li\u003e\n\u003cli\u003eB. Du, X. Jiang, A. Das, Q. Zhou, M. Yu, R. Jin, J. Zheng (2017) Glomerular barrier behaves as an atomically precise bandpass filter in a sub-nanometre regime, Nat. Nanotechnol. 12(11):1096-1102. https://doi.org/10.1038/nnano.2017.170\u003c/li\u003e\n\u003cli\u003eC.Q. Peng, Y.Y. Huang, J. Zheng (2020) Renal clearable nanocarriers: Overcoming the physiological barriers for precise drug delivery and clearance, J. Control. Release 322:64-80. https://doi.org/10.1016/j.jconrel.2020.03.020\u003c/li\u003e\n\u003cli\u003eI. Wessels, H.J. Fischer, L. Rink (2021) Dietary and Physiological Effects of Zinc on the Immune System, in: P.J. Stover, R. Balling (Eds.), Annu. Rev. Nutr. 41(1):133-175. https://doi.org/10.1146/annurev-nutr-122019-120635\u003c/li\u003e\n\u003cli\u003eA.U.R. Khan, K. Huang, M.S. Khalaji, F. Yu, X. Xie, T. Zhu, Y. Morsi, Z. Jinzhong, X. Mo (2021) Multifunctional bioactive core-shell electrospun membrane capable to terminate inflammatory cycle and promote angiogenesis in diabetic wound, Bioact. Mater. 6(9):2783-2800. https://doi.org/10.1016/j.bioactmat.2021.01.040\u003c/li\u003e\n\u003cli\u003eX. Yang, M. Jia, Z. Li, Z. Ma, J. Lv, D. Jia, D. He, R. Zeng, G. Luo, Y. Yu (2022) In-situ synthesis silver nanoparticles in chitosan/Bletilla striata polysaccharide composited microneedles for infected and susceptible wound healing, Int. J. of Biol. Macromol. 215:550-559. https://doi.org/10.1016/j.ijbiomac.2022.06.131\u003c/li\u003e\n\u003cli\u003eM.N. Liu, L. Zhang, R.R. Yang, H.Y. Cui, Y.A. Li, X.J. Li, H. Huang (2024) Integrating metal-organic framework ZIF-8 with green modifier empowered bacteria with improved bioremediation, J. Hazard. Mater. 461:132475. https://doi.org/10.1016/j.jhazmat.2023.132475\u003c/li\u003e\n\u003cli\u003eH. Akhtar, U. Amara, K. Mahmood, M. Hanif, M. Khalid, S. Qadir, Q.H. Peng, M. Safdar, M. Amjad, M.Z. Saif, A. Tahir, M. Yaqub, K. Khalid (2024) Drug carrier wonders: Synthetic strategies of zeolitic imidazolates frameworks (ZIFs) and their applications in drug delivery and anti-cancer activity, Adv. Colloid Interface Sci. 329:103184. https://doi.org/10.1016/j.cis.2024.103184\u003c/li\u003e\n\u003cli\u003eS. Yang, Y. Zhu, C. Ji, H. Zhu, A. Lao, R. Zhao, Y. Hu, Y. Zhou, J. Zhou, K. Lin, Y. Xu (2024) A five-in-one novel MOF-modified injectable hydrogel with thermo-sensitive and adhesive properties for promoting alveolar bone repair in periodontitis: Antibacterial, hemostasis, immune reprogramming, pro-osteo-/angiogenesis and recruitment, Bioact. Mater. 41:239-256. https://doi.org/10.1016/j.bioactmat.2024.07.016\u003c/li\u003e\n\u003cli\u003eH. Zhang, S. Yuan, B. Zheng, P. Wu, X. He, Y. Zhao, Z. Zhong, X. Zhang, J. Guan, H. Wang, L. Yang, X. Zheng (2025) Lubricating and Dual-Responsive Injectable Hydrogels Formulated From ZIF-8 Facilitate Osteoarthritis Treatment by Remodeling the Microenvironment, Small 21(3):2407885. https://doi.org/10.1002/smll.202407885\u003c/li\u003e\n\u003cli\u003eF. Guan, R.X. Wang, Z.J. Yi, P. Luo, W.Y. Liu, Y. Xie, Z.Q. Liu, Z.W. Xia, H. Zhang, Q. Cheng (2025) Tissue macrophages: origin, heterogenity, biological functions, diseases and therapeutic targets, Signal Transduct. Target. Ther. 10(1):93. https://doi.org/10.1038/s41392-025-02124-y\u003c/li\u003e\n\u003cli\u003eN. Eslahi, F. Soleimani, R. Lotfi, F. Mohandes, A. Simchi, M. Razavi (2024) How biomimetic nanofibers advance the realm of cutaneous wound management: The state-of-the-art and future prospects, Prog. Mater. Sci. 145:101293. \u003cu\u003ehttps://doi.org/10.1016/j.pmatsci.2024.101293\u003c/u\u003e\u003c/li\u003e\n\u003cli\u003eW.L. Liang, J.L. Cheng, J.D. Zhang, Q.Y. Xiong, M.J. Jin, J.H. Zhao (2022) pH-Responsive On-Demand Alkaloids Release from Core-Shell ZnO@ZIF-8 Nanosphere for Synergistic Control of Bacterial Wilt Disease, ACS Nano 16(2):2762-2773. https://doi.org/10.1021/acsnano.1c09724\u003c/li\u003e\n\u003cli\u003eM.H. Ali, M.A.K. Azad, K.A. Khan, M.O. Rahman, U. Chakma, A. Kumer (2023) Analysis of Crystallographic Structures and Properties of Silver Nanoparticles Synthesized Using PKL Extract and Nanoscale Characterization Techniques, ACS Omega 8(31):28133-28142. https://doi.org/10.1021/acsomega.3c01261\u003c/li\u003e\n\u003cli\u003eS. Sun, X. Meng, J. Xu, Z. Yang, X. Zhang, H. Dong (2024) Ag-decorated multivariate metal-organic frameworks for enhanced sonodynamic therapy of bacterial infections in wound healing, Nano Today 55:102181. https://doi.org/10.1016/j.nantod.2024.102181\u003c/li\u003e\n\u003cli\u003eZ. Li, J. Li (2023) Synergistic surface modulation of Ag/ZnO@ZIF-8 hybrid microspheres for enhanced photocatalytic degradation efficiency of Rhodamine B, CrystEngComm 25(14):2064-2074. https://doi.org/10.1039/D2CE01700H\u003c/li\u003e\n\u003cli\u003eL.-X. Liang, J.-C. Li, J.-W. Su, W.-W. Yang, Q. Xu, Z.-G. Qu (2025) ZIF-modified tailored multiscale-pore N-doped graphite felt for high performance vanadium redox flow batteries, J. Energy Storage 128:117252. https://doi.org/10.1016/j.est.2025.117252\u003c/li\u003e\n\u003cli\u003eJ. Cui, D. Wu, Z. Li, G. Zhao, J. Wang, L. Wang, B. Niu (2021) Mesoporous Ag/ZnO hybrid cages derived from ZIF-8 for enhanced photocatalytic and antibacterial activities, Ceram. Int. 47(11):15759-15770. https://doi.org/10.1016/j.ceramint.2021.02.148\u003c/li\u003e\n\u003cli\u003eN.J. Waleng, T.S. Munonde, A. Mpupa, T. Moremedi, Y. Zhang, P.N. Nomngongo (2025) Photoelectrochemical properties of magnetic amine-based MIL-101(Cr) hybrid material and its application in the degradation of acebutolol in water, RSC Adv. 15(23):17986-17999. https://doi.org/10.1039/D5RA02767E\u003c/li\u003e\n\u003cli\u003eY.T. Wen, N. Qi, Y. Zhang, Z.Q. Li, B. Fan, M. Zhang, Y. Li (2025)Green synthesis of polysaccharide-based Ag@AgCl photoactive porous hydrogel spheres for enhanced visible light photocatalytic degradation of organic pollutants, Carbohydr. Polym. 356:123401. https://doi.org/10.1016/j.carbpol.2025.123401\u003c/li\u003e\n\u003cli\u003eS.S. Hu, M.L. Gao, J.J. Huang, H. Wang, Q.Y. Wang, W.J. Yang, Z.H. Sun, X.S. Zheng, H.L. Jiang (2024) Introducing Hydrogen-Bonding Microenvironment in Close Proximity to Single-Atom Sites for Boosting Photocatalytic Hydrogen Production, J. Am. Chem. Soc. 146(29):20391-20400. https://doi.org/10.1021/jacs.4c06013\u003c/li\u003e\n\u003cli\u003eH. Yuan, H. Shimotani, J. Ye, S. Yoon, H. Aliah, A. Tsukazaki, M. Kawasaki, Y. Iwasa (2010)Electrostatic and electrochemical nature of liquid-gated electric-double-layer transistors based on oxide semiconductors, J. Am. Chem. Soc. 132(51):18402-18407. https://doi.org/10.1021/ja108912x\u003c/li\u003e\n\u003cli\u003eX. Huang, X. Li, C. Deng, X. Deng, Y. Qu, W. Yu, S. Wang, H. Du, S. Zhang, P. Zhang, X. Yue, W. Yu (2025) Fabrication of highly efficient ZnO-Pt catalysts assisted by biomass-derived carboxymethyl cellulose for the photodegradation of diverse antibiotics, J. Environ. Manage. 382:125418. https://doi.org/10.1016/j.jenvman.2025.125418\u003c/li\u003e\n\u003cli\u003eC.Y. Jiang, M.M. Sun, Y.S. Wang, C.X. Dong, Y. Yu, G. Wang, Y. Lu, Z.B. Chen (2025)Coordination Engineering in Fe-Mn Dual-Atom Nanozyme: Yielding ROS Storm to Efficiently Promote Wound Healing, Adv. Funct. Mater. 2424599. https://doi.org/10.1002/adfm.202424599\u003c/li\u003e\n\u003cli\u003eY. Hao, Y. Zhao, C. Zhang, X. Chen, N. Sun, W. Wei (2024) ZIF-8 derived Ag/ZnO photocatalyst with enriched Ag-ZnO interface for effective oxidation of methane to liquid oxygenates with simultaneous mmol-scale productivity and \u0026sim;100 % selectivity, Appl. Catal. B Environ. 359:124505. https://doi.org/10.1016/j.apcatb.2024.124505\u003c/li\u003e\n\u003cli\u003eY. Zhao, S. Zhao, Y. Du, Z. Gao, Y. Li, H. Ma, H. Li, X. Ren, Q. Fan, D. Wu, Q. Wei (2025)Inverse Oxide/Alloy-Structured Nanozymes with NIR-Triggered Enzymatic Cascade Regulation of ROS Homeostasis for Efficient Wound Healing, Adv. Mater. 37(14):2418731. https://doi.org/10.1002/adma.202418731\u003c/li\u003e\n\u003cli\u003eP. Hu, Z. Jia, S. Zhao, K. Lin, G. Yang, W. Guo, S. Yu, J. Cheng, G. Du, J. Shi (2024) Injectable Therapeutic Hydrogel with H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e Self-Supplying and GSH Consumption for Synergistic Chemodynamic/Low-Temperature Photothermal Inhibition of Postoperative Tumor Recurrence and Wound Infection, Adv. Healthc. Mater. 13(26):2401551. https://doi.org/10.1002/adhm.202401551\u003c/li\u003e\n\u003cli\u003eY.L. Lin, X.J. He, T. Huang, J. Zhao, L.Y. Liu, J.Q. He, J.L. Shen, Q.Z. Ren (2024) High-Valent Silver-Porphyrin complex hybrid graphene oxide nanoplatform promotes MRSA-Infected wound healing, Chem. Eng. J. 483:149279. https://doi.org/10.1016/j.cej.2024.149279\u003c/li\u003e\n\u003cli\u003eT. Nakazono, A.R. Parent, K. Sakai (2015) Improving singlet oxygen resistance during photochemical water oxidation by cobalt porphyrin catalysts, Chemistry 21(18):6723-6726. https://doi.org/10.1002/chem.201500716\u003c/li\u003e\n\u003cli\u003eL. Xiong, H. Qi, S. Zhang, L. Zhang, X. Liu, A. Wang, J. Tang (2023) Highly Selective Transformation of Biomass Derivatives to Valuable Chemicals by Single-Atom Photocatalyst Ni/TiO\u003csub\u003e2\u003c/sub\u003e, Adv. Mater. 35(16):2209646. https://doi.org/10.1002/adma.202209646\u003c/li\u003e\n\u003cli\u003eK.L. Wu, D.D. Zhu, X.L. Dai, W.N. Wang, X.Y. Zhong, Z.B. Fang, C. Peng, X.W. Wei, H.S. Qian, X.L. Chen, X.W. Wang, Z.B. Zha, L. Cheng (2022) Bimetallic oxide Cu\u003csub\u003e1.5\u003c/sub\u003eMn\u003csub\u003e1.5\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e cage-like frame nanospheres with triple enzyme-like activities for bacterial-infected wound therapy, Nano Today 43:101380. https://doi.org/10.1016/j.nantod.2022.101380\u003c/li\u003e\n\u003cli\u003eR. Łyżeń, G. Gawron, L. Kadziński, B. Banecki (2022) GSH Protects the Escherichia coli Cells from High Concentrations of Thymoquinone, Molecules 27(8):2546. https://doi.org/10.3390/molecules27082546\u003c/li\u003e\n\u003cli\u003eY.L. Zhao, Y. Wu, Q. Xu, Y. Liu, Z.Y. Song, H.Y. Han (2024) H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e self-supplying and GSH-depleting nanosystem for amplified NIR mediated-chemodynamic therapy of MRSA biofilm-associated infections, J. Nanobiotechnol. 22(1):117. https://doi.org/10.1186/s12951-024-02350-6\u003c/li\u003e\n\u003cli\u003eY.L. Fan, H.J. Liu, Z.L. Wang, Z.L. Wang, L.L. Zhu, Y.Z. Wang, F. Song (2024) A One-Nano MOF-Two-Functions Strategy Toward Self-healing, Anti-inflammatory, and Antibacterial Hydrogels for Infected Wound Repair, Chem. Eng. J. 497:155037. https://doi.org/10.1016/j.cej.2024.155037\u003c/li\u003e\n\u003cli\u003eH. Wang, M. Wang, X. Xu, P. Gao, Z. Xu, Q. Zhang, H. Li, A. Yan, R.Y. Kao, H. Sun (2021)Multi-target mode of action of silver against Staphylococcus aureus endows it with capability to combat antibiotic resistance, Nat. Commun. 12(1):3331. https://doi.org/10.1038/s41467-021-23659-y\u003c/li\u003e\n\u003cli\u003eS.B. Guo, X. Liu, H.H. Chen, J.W. Wang, Y.M. Qiao, T.L. Zhang, X.H. Ji, H. Han, Z.F. Liu, Y. Bai, J. Tang (2024) Antibacterial effect of the metal nanocomposite on Escherichia coli, J. Hazard. Mater. 476:135149. https://doi.org/10.1016/j.jhazmat.2024.135149\u003c/li\u003e\n\u003cli\u003eE.M. Darby, E. Trampari, P. Siasat, M.S. Gaya, I. Alav, M.A. Webber, J.M.A. Blair (2023) Molecular mechanisms of antibiotic resistance revisited, Nat. Rev. Microbiol. 21(5):280-295. https://doi.org/10.1038/s41579-022-00820-y\u003c/li\u003e\n\u003cli\u003eE.Z. Zhou, Y. Yang, J.L. Li, G.Y. Ren, L.K. Li, Z. Li, Y.P. Lu, T.Y. Gu, F.H. Wang, D.K. Xu (2025) A Novel Cu and Ag Bearing High-Entropy Alloy With Remarkable Antibacterial, Antiviral and Anti-Microbial Corrosion Performances, Adv. Funct. Mater. 35(15):2419467. https://doi.org/10.1016/j.jmst.2020.01.039\u003c/li\u003e\n\u003cli\u003eL. Tang, S. Xie, D. Wang, Y. Wei, X. Ji, Y. Wang, N. Zhao, Z. Mou, B. Li, W.R. Sun, P.Y. Wang, N.P. Basmadji, J.L. Pedraz, C. Vairo, E.G. Lafuente, M. Ramalingam, X. Xiao, R. Wang (2025) Astragalus polysaccharide/carboxymethyl chitosan/sodium alginate based electroconductive hydrogels for diabetic wound healing and muscle function assessment, Carbohydr. Polym. 350:123058. https://doi.org/10.1016/j.carbpol.2024.123058\u003c/li\u003e\n\u003cli\u003eL. Pang, C. Dai, L. Bi, Z. Guo, J. Fan (2017) Biosafety and Antibacterial Ability of Graphene and Graphene Oxide In Vitro and In Vivo, Nanoscale Res. Lett. 12(1):564. https://doi.org/10.1186/s11671-017-2317-0\u003c/li\u003e\n\u003cli\u003eY. Zhang, X.B. Wei, Y. Xu, W.D. Xia, C.Y. Zheng, H.Y. Zhang, W.T. Chen, K. Xu, Q. Huang (2025) Zinc sulfate gel reshapes the wound microenvironment to promote full-thickness wound healing in mice, Regen. Ther. 28:582-590. https://doi.org/10.1016/j.reth.2025.01.022\u003c/li\u003e\n\u003cli\u003eM.J. Li, J. Tian, K.K. Yu, H. Liu, X.Q. Yu, N. Wang, Q.M. Gong, K. Li, Y. Shen, X. Wei (2024) A ROS-responsive hydrogel incorporated with dental follicle stem cell-derived small extracellular vesicles promotes dental pulp repair by ameliorating oxidative stress, Bioact. Mater. 36:524-540. https://doi.org/10.1016/j.bioactmat.2024.06.036\u003c/li\u003e\n\u003cli\u003eW.J. Xiu, L. Wan, K.L. Yang, X. Li, L.H. Yuwen, H. Dong, Y.B. Mou, D.L. Yang, L.H. Wang (2022) Potentiating hypoxic microenvironment for antibiotic activation by photodynamic therapy to combat bacterial biofilm infections, Nat. Commun. 13(1):3875. https://doi.org/10.1038/s41467-022-31479-x\u003c/li\u003e\n\u003cli\u003eF.J. Shariatzadeh, S. Currie, S. Logsetty, R. Spiwak, S. Liu (2025) Enhancing wound healing and minimizing scarring: A comprehensive review of nanofiber technology in wound dressings, Prog. Mater. Sci. 147:101350. https://doi.org/10.1016/j.pmatsci.2024.101350\u003c/li\u003e\n\u003cli\u003eL. Zhang, Y.F. Lin, L. Hu, Y.N. Wang, C.H. Hu, X.Y. Shangguan, S.Z. Tang, J.C. Chen, P. Hu, Z.S. Chen, Z.F. Ke, Z. Chen (2025)Transient intracellular expression of PD-L1 and VEGFR2 bispecific nanobody in cancer cells inspires long-term T cell activation and infiltration to combat tumor and inhibit cancer metastasis, Mol. Cancer 24(1):1-28. https://doi.org/10.1186/s12943-025-02253-6\u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Zinc-silver composite material, Antibacterial, Antibacterial mechanism, Wound healing","lastPublishedDoi":"10.21203/rs.3.rs-7961947/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-7961947/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eThe treatment of wounds caused by drug-resistant bacteria is a significant challenge in clinical medicine. Silver (Ag) nanoparticles have attracted considerable attention due to their ability to inhibit drug-resistant bacteria. Nevertheless, the propensity of Ag nanoparticles to aggregate as well as their elevated toxicity have restricted their practical application. To address this issue, this study involved the design of a core-shell-type ZnO@ZIF-8/Ag nanocomposite material that combines high antibacterial activity with excellent biocompatibility. The mean diameter of the Ag nanoparticles in this material was approximately 2.4 nm, and they were highly dispersed. Within the wound microenvironment medium, antibacterial factors, such as hydroxyl radicals (\u0026middot;OH), singlet oxygen (\u003csup\u003e1\u003c/sup\u003eO\u003csub\u003e2\u003c/sub\u003e), Zn\u003csup\u003e2+\u003c/sup\u003e, and Ag\u003csup\u003e+\u003c/sup\u003e, were generated. The material induced bacterial death by altering the secondary structure of the cell wall of drug-resistant bacteria, thereby inhibiting respiration, lysing phospholipid layers, causing cellular content leakage, and disrupting \u003cem\u003eβ\u003c/em\u003e-lactamases. The introduction of zinc oxide (ZnO) significantly reduced the toxicity of the Ag nanoparticles and regulated macrophage polarization, which inhibited the secretion of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) by M1-type macrophages (M1) while concomitantly promoting the secretion of interleukin-10 (IL-10) and vascular endothelial growth factor (VEGF) by M2-type macrophages (M2). The expression of platelet-endothelial cell adhesion molecule-1 (CD31), type I collagen/fibronectin (COL-Ⅰ/FN), and proliferating cell nuclear antigen (PCNA) was significantly promoted, which significantly enhanced wound healing in infected wounds. This study thus offers a novel strategy for developing therapies against drug-resistant bacterial infections with the potential for clinical application.\u003c/p\u003e","manuscriptTitle":"A ZnO@ZIF-8/Ag Dual-Platform Nanocomposite Synergizing ROS Generation for Bactericidal-Immunomodulatory Wound Therapy","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2026-02-12 14:48:48","doi":"10.21203/rs.3.rs-7961947/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"36ceff75-9563-498d-9522-f787e2052346","owner":[],"postedDate":"February 12th, 2026","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2026-02-25T11:53:07+00:00","versionOfRecord":[],"versionCreatedAt":"2026-02-12 14:48:48","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-7961947","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-7961947","identity":"rs-7961947","version":["v1"]},"buildId":"XKTyCvWXoU3ODBz1xrDgd","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}