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Bismuth Tungstate−Silver Sulfide Z‑Scheme Heterostructure Nanoglue Promotes Wound Healing through Wound Sealing and Bacterial Inactivation Liqi Wei, Yining Chen, Xinru Yu, Yan Yan, Hongxiang Liu, Xingyu Cui, Xin Liu, Xiaodong Yang, Jiaqi Meng, Shuo Yang, Lili Wang, Xizhen Yang, Rui Chen,* and Yan Cheng* Cite This: ACS Appl. Mater. Interfaces 2022, 14, 53491−53500 Read Online ACCESS Metrics & More Article Recommendations *sı Supporting Information ABSTRACT: Rapid wound closure and bacterial inactivation are effective strategies to promote wound healing. Herein, a versatile nanoglue, bismuth tungstate (Bi2WO6)−silver sulfide (Ag2S) direct Z- scheme heterostructure nanoparticles (BWOA NPs), was designed to accelerate wound healing. BWOA NPs’ hollow structure and rough surface could effectively close wound tissues acting as a barrier between external bacteria and the wound. More importantly, the unique Z-scheme heterostructure endows BWOA NPs with an effective electron and hole separating ability with potent redox potential, where electrons and holes could effectively react with water and oxygen to produce reactive oxygen species, leading to a higher antibacterial activity against both endogenous and external bacteria at the wound site. A series of in vitro and in vivo biological assessments demonstrated that BWOA NPs could rapidly close wounds and promote wound healing. With sunlight irradiation, the inhibiting rates of BWOA NPs against Escherichia coli and Staphylococcus aureus are 61.62 ± 2.85 and 73.40 ± 3.28%, respectively. Also, the wound healing rate in BWOA NP-treated mice is 25.90 ± 5.85% higher than PBS. This design provides a new effective strategy to promote bacterial inactivation and accelerate wound healing. KEYWORDS: wound healing, Z-scheme, antibacterial, adhesion, reactive oxygen species ■ INTRODUCTION The skin is an effective barrier protecting the human body from noxious outside environmental threats.1 Once the skin is severely damaged and long exposed to harmful bacteria and viruses, it will lead to diseases such as ulceration, infection, sepsis, or even death.2−5 Thus, cutaneous wound healing is very important for the repair and recovery of tissue function after injury.6 Recently, certain nanomaterials with particular physical and chemical properties have been used for treating cutaneous wounds and accelerate their healing.7−10 Photo- catalytic antibacterial agents, such as titania,11 graphene quantum dots,12 copper metal−organic framework hydro- gels,13,14 and black phosphorus,15,16 have been widely investigated for their potential to accelerate the wound healing process due to their excellent photocatalytic antibacterial activity. However, the ideal nanomaterials should promote wound healing through various activities. Since bleeding and bacterial infection are characteristics in the early stage of wound formation, nanomaterials exhibiting hemostatic (stop bleeding) and antibacterial properties could be beneficial for speeding up wound healing. Rapid wound closure is an effective wound healing strategy because not only can it stop bleeding but also reduce the risk of bacterial infection due to decreased wound exposure. Some adhesives, such as bioinspired medical adhesives,17,18 self- assembling peptides,19 multifunctional films,20 or polymer- based hydrogels,21,22 have been designed to this effect. Recently, a few inorganic nanomaterials, such as iron oxide and ceria, have been used for wound closure as well due to their nanobridging effects;23,24 because of the excellent adhesive performance of biological tissues by silica nano- particles (NPs), which was reported by Rose et al.,25 our recent work has shown that a hollow and rough surface can effectively increase the adhesive performance of ceria NPs.26 Thus, we speculate that other inorganic NPs with similar morphology could be used for rapid wound closure. Received: August 25, 2022 Accepted: November 10, 2022 Published: November 23, 2022 Research Articlewww.acsami.org © 2022 American Chemical Society 53491 https://doi.org/10.1021/acsami.2c15299 ACS Appl. Mater. Interfaces 2022, 14, 53491−53500 D ow nl oa de d vi a U N IV R E G I D O C A R IR I on J an ua ry 2 7, 2 02 6 at 2 0: 29 :4 2 (U T C ). Se e ht tp s: //p ub s. ac s. or g/ sh ar in gg ui de lin es f or o pt io ns o n ho w to le gi tim at el y sh ar e pu bl is he d ar tic le s. https://pubs.acs.org/action/doSearch?field1=Contrib&text1="Liqi+Wei"&field2=AllField&text2=&publication=&accessType=allContent&Earliest=&ref=pdf https://pubs.acs.org/action/doSearch?field1=Contrib&text1="Yining+Chen"&field2=AllField&text2=&publication=&accessType=allContent&Earliest=&ref=pdf https://pubs.acs.org/action/doSearch?field1=Contrib&text1="Xinru+Yu"&field2=AllField&text2=&publication=&accessType=allContent&Earliest=&ref=pdf https://pubs.acs.org/action/doSearch?field1=Contrib&text1="Yan+Yan"&field2=AllField&text2=&publication=&accessType=allContent&Earliest=&ref=pdf https://pubs.acs.org/action/doSearch?field1=Contrib&text1="Hongxiang+Liu"&field2=AllField&text2=&publication=&accessType=allContent&Earliest=&ref=pdf https://pubs.acs.org/action/doSearch?field1=Contrib&text1="Xingyu+Cui"&field2=AllField&text2=&publication=&accessType=allContent&Earliest=&ref=pdf https://pubs.acs.org/action/doSearch?field1=Contrib&text1="Xin+Liu"&field2=AllField&text2=&publication=&accessType=allContent&Earliest=&ref=pdf https://pubs.acs.org/action/doSearch?field1=Contrib&text1="Xiaodong+Yang"&field2=AllField&text2=&publication=&accessType=allContent&Earliest=&ref=pdf https://pubs.acs.org/action/doSearch?field1=Contrib&text1="Jiaqi+Meng"&field2=AllField&text2=&publication=&accessType=allContent&Earliest=&ref=pdf https://pubs.acs.org/action/doSearch?field1=Contrib&text1="Jiaqi+Meng"&field2=AllField&text2=&publication=&accessType=allContent&Earliest=&ref=pdf https://pubs.acs.org/action/doSearch?field1=Contrib&text1="Shuo+Yang"&field2=AllField&text2=&publication=&accessType=allContent&Earliest=&ref=pdf https://pubs.acs.org/action/doSearch?field1=Contrib&text1="Lili+Wang"&field2=AllField&text2=&publication=&accessType=allContent&Earliest=&ref=pdf https://pubs.acs.org/action/doSearch?field1=Contrib&text1="Xizhen+Yang"&field2=AllField&text2=&publication=&accessType=allContent&Earliest=&ref=pdf https://pubs.acs.org/action/doSearch?field1=Contrib&text1="Rui+Chen"&field2=AllField&text2=&publication=&accessType=allContent&Earliest=&ref=pdf https://pubs.acs.org/action/doSearch?field1=Contrib&text1="Yan+Cheng"&field2=AllField&text2=&publication=&accessType=allContent&Earliest=&ref=pdf https://pubs.acs.org/action/showCitFormats?doi=10.1021/acsami.2c15299&ref=pdf https://pubs.acs.org/doi/10.1021/acsami.2c15299?ref=pdf https://pubs.acs.org/doi/10.1021/acsami.2c15299?goto=articleMetrics&ref=pdf https://pubs.acs.org/doi/10.1021/acsami.2c15299?goto=recommendations&?ref=pdf https://pubs.acs.org/doi/10.1021/acsami.2c15299?goto=supporting-info&ref=pdf https://pubs.acs.org/doi/10.1021/acsami.2c15299?fig=abs1&ref=pdf https://pubs.acs.org/toc/aamick/14/48?ref=pdf https://pubs.acs.org/toc/aamick/14/48?ref=pdf https://pubs.acs.org/toc/aamick/14/48?ref=pdf https://pubs.acs.org/toc/aamick/14/48?ref=pdf www.acsami.org?ref=pdf https://pubs.acs.org?ref=pdf https://pubs.acs.org?ref=pdf https://doi.org/10.1021/acsami.2c15299?urlappend=%3Fref%3DPDF&jav=VoR&rel=cite-as https://www.acsami.org?ref=pdf https://www.acsami.org?ref=pdf Bacterial inactivation is another effective strategy to promote wound healing.27 Inorganic nanomaterials, especially semi- conductor nanomaterials, which can produce photoinduced electrons and holes to generate reactive oxygen species (ROS) by light irradiation, have been used as antibacterial agents to promote wound healing.28,29 However, their antibacterial efficacy has been limited by the fast recombination of photoinduced electrons and holes. Forming heterostructures between two nanomaterials, especially Z-scheme heterostruc- tures,is a proven effective way to improve their ROS generation ability since Z-scheme heterostructures could not only separate photoinduced electrons and holes based on step- wise energy levels but also better maintain the strong redox potential of separated electrons and holes compared with traditional type I and II heterostructures.30,31 Bismuth tungstate (Bi2WO6, abbreviated as BWO) nano- materials have been used in tumor therapy32,33 and photo- electric chemical immunosensors34 with excellent biocompat- ibility. As an N-type semiconductor with a bandgap of ∼2.77 eV,35,36 Bi2WO6 can absorb lightBWO were deduced to be −0.69 eV (0.23 eV) and −0.17 eV (2.63 eV), respectively. Therefore, when BWO hybridizes with Ag2S, the electrons in Ag2S would transfer to BWO due to the Fermi equilibrium, leading to a built-in electric field pointing Figure 2. Physicochemical characterization of BWOA NPs. (a) TEM image of BWO NPs. (b) TEM and HRTEM images of BWOA NPs. (c) STEM and element mapping images of BWOA NPs. (d) XRD patterns and (e) UV−Vis diffuse reflectance spectra of BWO and BWOA NPs. Figure 3. Energy band structures of BWO and Ag2S as well as the formation of a BWOA Z-scheme heterostructure. (a, b) Kubelka−Munk function deduced from the UV−Vis diffuse reflectance spectra of Ag2S and BWO NPs. (c) Mott−Schottky plots and (d) XPS valence spectra of Ag2S and BWO NPs. (e) Ag 3d, (f) Bi 4f, and (g) W 4f high-resolution XPS spectra of Ag2S, BWO, and BWOA NPs. (h) Scheme of energy band structures of Ag2S and BWO NPs and the formation of the BWOA Z-scheme heterostructure. ACS Applied Materials & Interfaces www.acsami.org Research Article https://doi.org/10.1021/acsami.2c15299 ACS Appl. Mater. Interfaces 2022, 14, 53491−53500 53493 https://pubs.acs.org/doi/suppl/10.1021/acsami.2c15299/suppl_file/am2c15299_si_001.pdf https://pubs.acs.org/doi/suppl/10.1021/acsami.2c15299/suppl_file/am2c15299_si_001.pdf https://pubs.acs.org/doi/suppl/10.1021/acsami.2c15299/suppl_file/am2c15299_si_001.pdf https://pubs.acs.org/doi/suppl/10.1021/acsami.2c15299/suppl_file/am2c15299_si_001.pdf https://pubs.acs.org/doi/suppl/10.1021/acsami.2c15299/suppl_file/am2c15299_si_001.pdf https://pubs.acs.org/doi/10.1021/acsami.2c15299?fig=fig2&ref=pdf https://pubs.acs.org/doi/10.1021/acsami.2c15299?fig=fig2&ref=pdf https://pubs.acs.org/doi/10.1021/acsami.2c15299?fig=fig2&ref=pdf https://pubs.acs.org/doi/10.1021/acsami.2c15299?fig=fig2&ref=pdf https://pubs.acs.org/doi/10.1021/acsami.2c15299?fig=fig3&ref=pdf https://pubs.acs.org/doi/10.1021/acsami.2c15299?fig=fig3&ref=pdf https://pubs.acs.org/doi/10.1021/acsami.2c15299?fig=fig3&ref=pdf https://pubs.acs.org/doi/10.1021/acsami.2c15299?fig=fig3&ref=pdf www.acsami.org?ref=pdf https://doi.org/10.1021/acsami.2c15299?urlappend=%3Fref%3DPDF&jav=VoR&rel=cite-as from Ag2S to BWO. Furthermore, the high-resolution XPS spectra of Ag 3d, Bi 4f, and W 4f were analyzed to confirm BWOA’s formation of a Z-scheme heterostructure. Figure 3e shows the high-resolution XPS spectra of Ag 3d, with binding energies at 373.95 and 367.9 eV in Ag2S NPs shown for Ag 3d 5/2 and Ag 3d 3/2, respectively, while in BWOA NPs, these peaks shift to lower binding energies (373.75 eV for Ag 3d 5/2 and 367.7 eV for Ag 3d 3/2). Meanwhile, the characteristic peaks of Bi 4f (Figure 3f) and W 4f (Figure 3g) shifted toward higher binding energies (0.1 eV for Bi 4f and 0.15 eV for W 4f). Generally, such binding energy shifts indicate a strong interaction between two different components in the heterostructure interface, implying the formation of a BWOA heterostructure. More importantly, these shifts indicate that the electrons would flow from Ag2S to BWOA, further confirming the formation of a built-in E-field pointing from Ag2S to BWOA. As shown in Figure 3h, the built-in E-field between Ag2S and BWO would promote photoinduced electron transfer from the CB of BWO to VB of Ag2S, proving the formation of BWOA’s Z-scheme heterostructure, which is similar to other Z-scheme heterostructures.23,50,51 As a result, BWO and Ag2S could form a Z-scheme heterostructure, which assists in the production of photoinduced electrons and holes with strong redox potential (Figure 3h). Tissue Adhesive Capability, Sunlight-Triggered ROS Generation, and Antibacterial Activity of BWOP Z- Scheme Heterostructure NPs. The tissue adhesive ability of BWO and BWOA NPs was detected by using them to attach two pieces of mouse skin. As shown in Figure 4a, both the physical mixture of BWO and Ag2S (BWO + A) NPs and BWOA NPs could adhere the two pieces of skin, while phosphate buffered saline (PBS, pH 7.4) could not, indicating the adhesive ability of both BWO + A and BWOA NPs. Moreover, BWOA NPs showed better adhesion than BWO + A NPs. To further quantify the adhesive effect of BWOA NPs, the displacement curve of BWO + A and BWOA NPs by the lap-shear adhesion test is displayed in Figure 4b, which shows the significantly larger failure force and displacement of both BWO + A and BWOA NPs compared to PBS, demonstrating the adhesive ability of these two NPs. In addition, the larger failure force and displacement of BWOA NPs also indicated the stronger adhesion ability of BWOA NPs, which may be due to BWOA’s coarser surface as shown by the TEM images (Figure 2a). As proven by the energy band structure (Figure 3h), the photoinduced electrons in the CB of Ag2S and photoinduced holes in the VB of BWO could react with oxygen and water to produce O2·− and OH·, respectively, due to their excellent separating ability of photoinduced electrons and holes. The photoluminescence, electrochemical impedance spectra (EIS), and photocurrent production of BWO and BWOA NPs served to investigate the separation ability of photoinduced electrons and holes. As shown in Figure 4c, with an excitation wavelength of 320 nm, BWO could produce an emission peak in the visible region at ∼520 nm, however, the intensity of this emission was weak in BWOA NPs, indicating the excellent electron−hole separation ability of Z-scheme heterostructures. Figure 4d and Figure S5 show the EIS spectra and photocurrents of BWO and BWOA NPs, in which BWOA showed a smaller semicircle in Nyquist plots and larger photocurrent than BWO, implying a more efficient charge separation of Z-scheme heterostructures. Next, the sunlight- Figure 4. In vitro wound healing ability of BWOA Z-scheme heterostructure NPs. (a) Photos and (b) force−displacement curves for lap joints of two pieces of mouse skin stuck by 2 μL of PBS, BWOA (100 mg mL−1), or BWO + A (90.6 mg mL−1 for BWO and 9.4 mg mL−1 for Ag2S). (c) PL and (d) EIS spectra of BWO and BWOA NPs. (e) XTT absorption and (f) APF fluorescence after incubation with PBS, Ag2S (9.4 μg mL−1), BWO (90.6 μg mL−1), BWOA (100 μg mL−1), and BWO + A (90.6 μg mL−1 for BWO and 9.4 μg mL−1 for Ag2S) with SSR (0.1 W cm−2, 15 min). ESR spectra of (g) DMPO-O2 •− methanol dispersion and (h) DMPO-OH• aqueous dispersion of Ag2S (9.4 μg mL−1), BWO (90.6 μg mL−1), and BWOA (100 μg mL−1) with SSR (0.1 W cm−2, 15 min). ACS Applied Materials & Interfaces www.acsami.org Research Article https://doi.org/10.1021/acsami.2c15299 ACS Appl. Mater. Interfaces 2022, 14, 53491−53500 53494 https://pubs.acs.org/doi/suppl/10.1021/acsami.2c15299/suppl_file/am2c15299_si_001.pdf https://pubs.acs.org/doi/10.1021/acsami.2c15299?fig=fig4&ref=pdf https://pubs.acs.org/doi/10.1021/acsami.2c15299?fig=fig4&ref=pdf https://pubs.acs.org/doi/10.1021/acsami.2c15299?fig=fig4&ref=pdf https://pubs.acs.org/doi/10.1021/acsami.2c15299?fig=fig4&ref=pdf www.acsami.org?ref=pdf https://doi.org/10.1021/acsami.2c15299?urlappend=%3Fref%3DPDF&jav=VoR&rel=cite-as triggered ROS generation ability of BWOA Z-scheme heterostructure NPs was investigated by 3′-(p-aminophenyl) fluorescein (APF) and 2,3-bis-(2-methoxy-4-nitro-5-sulfopheh- yl)-2H-tetrazolium-5-carboxanilide) (XTT), which produces fluorescence and absorbance when reacting with OH· and O2·−, respectively. As shown in Figure 4e and Figure S6, with SSR, only BWOA NPs could induce XTT absorption at 470 nm, while no absorption was detected in either sunlight- triggered BWO, Ag2S, and BWO + A NPs or in any of these NPs without SSR, meaning that, due to their stronger reduction ability, the photoinduced electrons in BWOA Z- scheme heterostructures could react with O2 to generate O2·−. Meanwhile, without SSR, no APF fluorescence was generated when incubating with PBS, Ag2S, BWO, BWO + A, or BWOA NPs (Figure S7), while BWOA NPs could produce more APF fluorescence with SSR than BWO + A NPs (Figure 4f), implying that the photoinduced holes in BWOA Z-scheme heterostructures could reactwith water to generate OH·. In addition, the generation of O2·− and OH· species was further verified by electron spin resonance (ESR) spectra. As shown in Figures S8 and S9, no ESR signal was detected without SSR. With SSR, both O2·− and OH· were detected in BWOA NPs, in which BWOA’s signal intensity was higher than those of BWO and Ag2S (Figure 4g,h). These results further demonstrated the excellent O2·− and OH· generation ability of BWOA Z-scheme heterostructures. Encouraged by the excellent ROS generation ability detected in our previous experiments, we investigated BWOA Z-scheme heterostructure NP’s antibacterial activity against both Gram- negative Escherichia coli (E. coli) and Gram-positive Staph- ylococcus aureus (S. aureus). PBS, Ag2S, BWO, BWO + A NPs, and commercial titanium dioxide (P25) were used as the control. As shown in Figure 5a, the growth of both Gram- negative and Gram-positive bacteria was significantly inhibited by BWOA NPs with SSR; the inhibiting efficacy was higher than those of other NPs, while bacterial growth was not inhibited without SSR (Figure S10). The quantitative analysis of bacterial growth based on the optical images of bacterial colonies (n = 3) showed inhibiting rates of BWOA NPs against E. coli and S. aureus of 61.62 ± 2.85% and 73.40 ± 3.28%, respectively (Figure S11), higher than those of BWO + A NPs (36.94 ± 5.34% for E. coli and 37.37 ± 2.50% for S. aureus). We also measured the Ag+ release in BWOA NPs by an inductively coupled plasma optical emission spectrometer, finding that only 3.43 μg mL−1 of Ag+ was released from the BWOA aqueous solution (600 μg mL−1). More importantly, the released Ag+ had no antibacterial activity as proven by antibacterial tests (Figure S9 and Figure 5a), showing that the antibacterial ability originated from BWOA Z-scheme heterostructures. Figure 5b and Figure S12 show the live/ dead staining of bacteria after BWO + A and BWOA NP treatment with SSR, which further confirmed the bacterial inactivation ability of BWOA Z-scheme heterostructure NPs. Figure 5. Antibacterial ability of BWOA Z-scheme heterostructure NPs. (a) Optical images of bacterial colonies of E. coli and S. aureus treated with PBS, Ag2S (56.4 μg mL−1), BWO (543.6 μg mL−1), BWOA (600 μg mL−1), BWO + A (543.6 μg mL−1 for BWO and 56.4 μg mL−1 for Ag2S), and P25 (600 μg mL−1) for 24 h with SSR (0.1 W cm−2, 15 min). Fluorescence images of (b) live/dead staining and (c) DCF staining (scale bar = 20 μm) of E. coli and S. aureus treated with PBS, BWOA (600 μg mL−1), and BWO + A (543.6 μg mL−1 for BWO and 56.4 μg mL−1 for Ag2S) for 6 h with SSR (0.1 W cm−2, 15 min). GSH levels of E. coli and S. aureus treated with PBS, BWOA (600 μg mL−1), and BWO + A (543.6 μg mL−1 for BWO and 56.4 μg mL−1 forAg2S) (d) with or (e) without SSR (0.1 W cm−2, 15 min) (n = 3, *pfacilitating highly antibacterial activity. As a result, BWOA Z-scheme heterostructure NPs can act as a multifunctional nanoglue, which holds great potential for accelerating the wound healing process based on their rapid wound closure and bacteria inactivation abilities. ■ EXPERIMENTAL SECTION Preparation of BWO and BWOA NPs. Bi2O3 templates were first synthesized through a hydrothermal method. In detail, urea (0.016 g) and polyvinylpyrrolidone (MW = 10,000, 0.6 g) were dissolved in ethylene glycol (50 mL), and then, nitric acid solution (6.28%, 10 mL) containing 0.364 g of bismuth(III) nitrate pentahydrate was added. After stirring for 30 min, the above solution was transferred into a 100 mL stainless steel autoclave, sealed, and heated at 150 °C for 3 h. Finally, Bi2O3 NPs were obtained by centrifugation, washing with deionized water, and drying at 60 °C. Next, the collected Bi2O3 NPs were dispersed into deionized water to form an aqueous solution (4.657 mg mL−1, 14 mL), to which 0.0462 g of sodium tungstate was added and stirred for 30 min. After being transferred into a 20 mL stainless steel autoclave and heated at 150 °C for 12 h, hollow BWO NPS were collected by centrifugation, washing, and drying. Finally, BWAO NPs were obtained by in situ growth of Ag2S on the surface of Bi2WO6 NPs. Typically, silver nitrate (0.0017 g) was added into BWO dispersions (1 mg mL−1, 30 mL) and stirred for 30 min, and then, sodium sulfide (0.0015 g) was added and stirred for another 2 h. The precipitates were collected by centrifugation and washed with deionized water three times. Finally, BWOA NPs were obtained after drying at 60 °C for 12 h. Figure 6. In vivo wound healing ability of BWOA Z-scheme heterostructure NPs. (a) Images of wounds before and after closure. The wounds were treated with 20 μL of PBS, BWOA (10 mg mL−1), and BWO + A (9.06 mg mL−1 for BWO and 0.94 mg mL−1 for Ag2S). (b) Quantification of wound healing rate expressed as percentage of the initial wound length (n = 5, **PAgricultural University. First, 1.5 cm-long transverse full-thickness cutting wounds were made on the back of mice after anesthetizing. Then, 20 μL of PBS, BWO + A (9.06 mg mL−1 for BWO and 0.94 mg mL−1 for Ag2S), or BWOA (10 mg mL−1) NPs were added to the wound site of mice, and both edges of wounds were pressed with fingers for 30 s and excessive NP solution was removed. The above mice were divided into two groups, one was irradiated with simulated sunlight (0.1 W cm−2) for 30 min, and the other was not. The wounds were measured and photographed every 2 days. After wound healing, the mice were sacrificed and wound tissues were embedded with paraffin for H&E and Masson’s trichrome staining. Statistical Analysis. 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