{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T01:33:35Z","timestamp":1760060015392,"version":"build-2065373602"},"reference-count":28,"publisher":"MDPI AG","issue":"15","license":[{"start":{"date-parts":[[2025,7,24]],"date-time":"2025-07-24T00:00:00Z","timestamp":1753315200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"MEXT","award":["22H03956","25K15914"],"award-info":[{"award-number":["22H03956","25K15914"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Materials"],"abstract":"<jats:p>Salmonella enterica serovar Typhimurium (S. Typhimurium) is an intracellular pathogen capable of surviving and replicating within macrophages, which causes foodborne diseases such as gastroenteritis. To develop a strategy against intracellular bacteria in macrophages, we designed silver-loaded biodegradable polymer nanoparticles functionalized with S. Typhimurium membrane vesicles (MVs). Silver nanoparticles (Ag NPs) were initially encapsulated within biodegradable poly(lactic-co-glycolic) nanoparticles (Ag-P NPs), which were then surface-modified with polyethyleneimine to form Ag-PP NPs. These were subsequently fused with S. Typhimurium MVs via a sonication method to generate Ag-PP@MV NPs. The resulting MV-coated nanoparticles displayed a similar protein profile to that of native MVs and exhibited antibacterial activity against intracellular S. Typhimurium. Notably, the enhanced cellular uptake of the MV-modified NPs contributed to their intracellular bactericidal efficacy. This study highlights MV modification as a promising strategy to improve NP delivery to macrophages for treating persistent intracellular infections.<\/jats:p>","DOI":"10.3390\/ma18153470","type":"journal-article","created":{"date-parts":[[2025,7,24]],"date-time":"2025-07-24T10:28:09Z","timestamp":1753352889000},"page":"3470","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Modification of Silver-Loaded Biodegradable Polymer Nanoparticles with Bacterial Membrane Vesicles for Treating Intracellular Bacterial Infections"],"prefix":"10.3390","volume":"18","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-1332-3189","authenticated-orcid":false,"given":"Wei","family":"Xu","sequence":"first","affiliation":[{"name":"Faculty of Advanced Science and Technology, Kumamoto University, 2-39-1 Kurokami, Chuo-ku, Kumamoto 860-8555, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sayo","family":"Maruyama","sequence":"additional","affiliation":[{"name":"Faculty of Advanced Science and Technology, Kumamoto University, 2-39-1 Kurokami, Chuo-ku, Kumamoto 860-8555, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8070-8708","authenticated-orcid":false,"given":"Takuro","family":"Niidome","sequence":"additional","affiliation":[{"name":"Faculty of Advanced Science and Technology, Kumamoto University, 2-39-1 Kurokami, Chuo-ku, Kumamoto 860-8555, Japan"},{"name":"Institute of Industrial Nanomaterials, Kumamoto University, 2-39-1 Kurokami, Chuo-ku, Kumamoto 860-8555, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2025,7,24]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"355","DOI":"10.1038\/nrmicro2128","article-title":"Antimicrobial mechanisms of phagocytes and bacterial evasion strategies","volume":"7","author":"Flannagan","year":"2009","journal-title":"Nat. Rev. Microbiol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"900","DOI":"10.1074\/mcp.RA119.001841","article-title":"Proteomic analysis of Salmonella-modified membranes reveals adaptations to macrophage hosts","volume":"18","author":"Reuter","year":"2020","journal-title":"Mol. Cell. Proteom."},{"key":"ref_3","doi-asserted-by":"crossref","unstructured":"Lamichhane, B., Mawad, A.M.M., Saleh, M., Kelley, W.G., Harrington, P.J., Lovestad, C.W., Amezcua, J., Sarhan, M.M., El Zowalaty, M.E., and Ramadan, H. (2024). Salmonellosis: An overview of epidemiology, pathogenesis, and innovative approaches to mitigate the antimicrobial resistant infections. Antibiotics, 13.","DOI":"10.3390\/antibiotics13010076"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"735","DOI":"10.2147\/IDR.S345574","article-title":"Clinical perspective of antimicrobial resistance in bacteria","volume":"15","author":"Zhu","year":"2022","journal-title":"Infect. Drug Resist."},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"Sridhar, S., Forrest, S., Pickard, D., Cormie, C., Lees, E.A., Thomson, N.R., Dougan, G., and Baker, S. (2021). Inhibitory Concentrations of Ciprofloxacin Induce an Adaptive Response Promoting the Intracellular Survival of Salmonella enterica Serovar Typhimurium. mBio, 12.","DOI":"10.1128\/mBio.01093-21"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"388","DOI":"10.1039\/D4PM00032C","article-title":"Antimicrobial nanoparticles: Current landscape and future challenges","volume":"1","author":"Mondal","year":"2024","journal-title":"RSC Pharm."},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Bamal, D., Singh, A., Chaudhary, G., Kumar, M., Singh, M., Rani, N., Mundlia, P., and Sehrawat, A.R. (2021). Silver Nanoparticles Biosynthesis, Characterization, Antimicrobial Activities, Applications, Cytotoxicity and Safety Issues: An Updated Review. Nanomaterials, 11.","DOI":"10.3390\/nano11082086"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"upae084","DOI":"10.1093\/chemle\/upae084","article-title":"Encapsilation of silver nanoplates into poly(lactic-co-glycolic) acid nanoparticles and the antibacterial activity against intracellular bacteria","volume":"54","author":"Nawata","year":"2024","journal-title":"Chem. Lett."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.colsurfb.2009.09.001","article-title":"Biodegradable polymeric nanoparticles based drug delivery systems","volume":"75","author":"Kumari","year":"2010","journal-title":"Colloids Surf. B Biointerfaces"},{"key":"ref_10","first-page":"749","article-title":"Systemic delivery to central nervous system by engineered PLGA nanoparticles","volume":"8","author":"Cai","year":"2016","journal-title":"Am. J. Transl. Res."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"2053","DOI":"10.1016\/S0142-9612(02)00641-5","article-title":"New micelle-like polymer aggregated made from PEI-PLGA deblock copolymers: Micellar characteristics and cellular uptake","volume":"24","author":"Nam","year":"2003","journal-title":"Biomaterials"},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Aytar \u00c7elik, P., Erdogan-Gover, K., Barut, D., Enuh, B.M., Amasya, G., Sengel-T\u00fcrk, C.T., Derkus, B., and \u00c7abuk, A. (2023). Bacterial Membrane Vesicles as Smart Drug Delivery and Carrier Systems: A New Nanosystems Tool for Current Anticancer and Antimicrobial Therapy. Pharmaceutics, 15.","DOI":"10.3390\/pharmaceutics15041052"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"5012","DOI":"10.1016\/j.vaccine.2015.05.014","article-title":"Altered host immune responses to membrane vesicles from Salmonella and Gram-negative pathogens","volume":"33","author":"Laughlin","year":"2015","journal-title":"Vaccine"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"7692","DOI":"10.4049\/jimmunol.179.11.7692","article-title":"Membrane vesicles are immunogenic facsimiles of salmonella typhimurium that potently activate dendritic cells, prime B and T cell responses and stimulate protective immunity in vivo","volume":"179","author":"Alaniz","year":"2007","journal-title":"J. Immunol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"2105854","DOI":"10.1002\/advs.202105854","article-title":"Biomimetic lipopolysaccharide-free bacterial outer membrane-functionalized nanoparticles for brain-targeted drug delivery","volume":"9","author":"Chen","year":"2022","journal-title":"Adv. Sci."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"2182","DOI":"10.1128\/IAI.01277-10","article-title":"Discovery of Salmonella Virulence Factors Translocated via Outer Membrane Vesicles to Murine Macrophages","volume":"79","author":"Yoon","year":"2011","journal-title":"Infect. Immun."},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Wu, N., Han, Z., Lv, W., Huang, Y., Zhu, J., Deng, J., and Xue, Q. (2025). Reprogramming peritoneal macrophages with outer membrane vesicles-coated PLGA nanoparticles for endometriosis prevention. Biomaterials, 319.","DOI":"10.1016\/j.biomaterials.2025.123198"},{"key":"ref_18","doi-asserted-by":"crossref","unstructured":"Xu, W., Maruyama, S., Sato, A., and Niidome, T. (2024). Bacterial membrane vesicles combined with nanoparticles for bacterial vaccines and cancer immunotherapy. Colloids Surf. B Biointerfaces, 243.","DOI":"10.1016\/j.colsurfb.2024.114125"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"5500","DOI":"10.1021\/jp031077n","article-title":"Silver nanoplates: Size control in two dimensions and formation mechanisms","volume":"108","author":"Chen","year":"2004","journal-title":"J. Phys. Chem. B"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"313081","DOI":"10.1155\/2013\/313081","article-title":"Detection limits of DLS and UV-Vis spectroscopy in characterization of polydisperse nanoparticles colloids","volume":"2013","author":"Tomaszewska","year":"2013","journal-title":"J. Nanomater."},{"key":"ref_21","doi-asserted-by":"crossref","unstructured":"Sharfalddin, A.A., Emwas, A.H., Jaremko, M., and Hussien, M.A. (2021). Complexation of uranyl (UO2)2+ with bidentate ligands: XRD, spectroscopic, computational, and biological studies. PLoS ONE, 16.","DOI":"10.1371\/journal.pone.0256186"},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"Szatanek, R., Baj-Krzyworzeka, M., Zimoch, J., Lekka, M., Siedlar, M., and Baran, J. (2017). The Methods of Choice for Extracellular Vesicles (EVs) Characterization. Int. J. Mol. Sci., 18.","DOI":"10.3390\/ijms18061153"},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Dumontel, B., Susa, F., Limongi, T., Vighetto, V., Debellis, D., Canta, M., and Cauda, V. (2022). Nanotechnological engineering of extracellular vesicles for the development of actively targeted hybrid nanodevices. Cell Biosci., 12.","DOI":"10.1186\/s13578-022-00784-9"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"2000015","DOI":"10.1002\/adfm.202000015","article-title":"Autologous versatile vesicles-incorporated biomimetic extracellular matrix induces biomineralization","volume":"30","author":"Wei","year":"2020","journal-title":"Adv. Funct. Mater."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"188","DOI":"10.1021\/acs.est.5b03982","article-title":"Rethinking stability of silver sulfide nanoparticles (Ag2S-NPs) in the aquatic environment: Photoinduced transformation of Ag2S-NPs in the presence of Fe(III)","volume":"50","author":"Li","year":"2016","journal-title":"Environ. Sci. Technol."},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Liu, Q., Liu, Q., Yi, J., Liang, K., Hu, B., Zhang, X., and Kong, Q. (2016). Outer membrane vesicles from flagellin-deficient Salmonella enterica serovar Typhimurium induce cross-reactive immunity and provide cross-protection against heterologous Salmonella challenge. Sci. Rep., 6.","DOI":"10.1038\/srep34776"},{"key":"ref_27","first-page":"522","article-title":"Induced fusion and aggregation of bacterial outer membrane vesicles: Experimental and theoretical analysis","volume":"578","author":"Gnopo","year":"2020","journal-title":"Colloids Surf. B Biointerfaces"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"6958","DOI":"10.1038\/s41467-024-51364-z","article-title":"A pH-sensitive motif in an outer membrane protein activates bacterial membrane vesicle production","volume":"15","author":"Dehinwal","year":"2024","journal-title":"Nat. Commun."}],"container-title":["Materials"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1996-1944\/18\/15\/3470\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,9]],"date-time":"2025-10-09T18:15:15Z","timestamp":1760033715000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1996-1944\/18\/15\/3470"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,7,24]]},"references-count":28,"journal-issue":{"issue":"15","published-online":{"date-parts":[[2025,8]]}},"alternative-id":["ma18153470"],"URL":"https:\/\/doi.org\/10.3390\/ma18153470","relation":{},"ISSN":["1996-1944"],"issn-type":[{"type":"electronic","value":"1996-1944"}],"subject":[],"published":{"date-parts":[[2025,7,24]]}}}