{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,26]],"date-time":"2026-08-26T08:21:05Z","timestamp":1787732465541,"version":"build-2784847793"},"reference-count":40,"publisher":"Wiley","issue":"4","license":[{"start":{"date-parts":[[2026,8,12]],"date-time":"2026-08-12T00:00:00Z","timestamp":1786492800000},"content-version":"vor","delay-in-days":11,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2026,8,1]],"date-time":"2026-08-01T00:00:00Z","timestamp":1785542400000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["MicrobiologyOpen"],"published-print":{"date-parts":[[2026,8]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:p>\n                    Human microbiota is increasingly considered to shape health and disease, drawing interest of pharma and biotech industries in advanced models of in vitro human microbiome to streamline drug development. In this context, Universal\u2010Bac\n                    <jats:sup>3<\/jats:sup>\n                    Gel represents a new generation of 3D biomaterials designed to mimic the properties of human mucus and biofilm features, including micro\u2010gradients that replicate the heterogeneous environments colonized by microorganisms in the human body. To evaluate the suitability of Universal\u2010Bac\n                    <jats:sup>3<\/jats:sup>\n                    Gel for studying clinically relevant species in antimicrobial resistance, the so\u2010called ESKAPE pathogens (\n                    <jats:italic>Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterobacter cloacae<\/jats:italic>\n                    ) were cultured within this 3D environment. Bacterial growth was monitored at 24\u2010 and 48\u2010h post\u2010inoculation via spot plating, while viability, spatial distribution, and organization were assessed by confocal laser scanning microscopy. All ESKAPE strains successfully grew throughout the structure of Universal\u2010Bac\n                    <jats:sup>3<\/jats:sup>\n                    Gel. Distinct 3D biofilm architectures were observed across species, ranging from diffuse colonization to compact microcolony formation, in agreement with species\u2010specific biofilm patterns. Ciprofloxacin susceptibility assays revealed reduced susceptibility of bacteria cultured within Universal\u2010Bac\n                    <jats:sup>3<\/jats:sup>\n                    Gel compared with their planktonic counterparts, supporting the development of biofilm\u2010associated tolerance phenotypes. Consistent with these findings, crystal violet staining confirmed the accumulation of biofilm\u2010associated biomass within the hydrogel. Notably, the platform's ready\u2010to\u2010use 96\u2010well format allowed direct comparison of these high\u2010priority pathogens under standardized conditions, highlighting species\u2010specific biofilm traits that would be difficult to discern in conventional two\u2010dimensional culture systems. This work highlights the versatility of Universal\u2010Bac\n                    <jats:sup>3<\/jats:sup>\n                    Gel as a biofilm\u2010relevant in vitro platform for studying pathogen colonization, biofilm development and antimicrobial susceptibility under controlled conditions.\n                  <\/jats:p>","DOI":"10.1002\/mbo3.70371","type":"journal-article","created":{"date-parts":[[2026,8,13]],"date-time":"2026-08-13T04:51:31Z","timestamp":1786596691000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Universal\u2010Bac\n                    <sup>3<\/sup>\n                    Gel: A 3D Biofilm\u2010Relevant Matrix That Supports In Vitro Growth and Biofilm Formation of ESKAPE Pathogens"],"prefix":"10.1002","volume":"15","author":[{"ORCID":"https:\/\/orcid.org\/0009-0009-2882-1613","authenticated-orcid":false,"given":"Emanuela","family":"Peluso","sequence":"first","affiliation":[{"name":"Molecular Medicine Department (DMM), Centre for Health Technologies (CHT), Unit\u00e0 di Ricerca (UdR) INSTM, Operative Unit (OU) of the Interuniversity Center for the Promotion of the 3Rs Principles in Teaching and Research (Centro 3R) University of Pavia Pavia Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9031-2371","authenticated-orcid":false,"given":"Sebasti\u00e3o","family":"van Uden","sequence":"additional","affiliation":[{"name":"Bac<sup>3<\/sup>Gel\u00ae Lda, Edif\u00edcio Inova\u00e7\u00e3o II Porto Salvo Portugal"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7262-6166","authenticated-orcid":false,"given":"Sonja","family":"Visentin","sequence":"additional","affiliation":[{"name":"Department of Molecular Biotechnology and Health Sciences University of Torino Turin Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5081-5254","authenticated-orcid":false,"given":"Paola","family":"Petrini","sequence":"additional","affiliation":[{"name":"BioAvatar Lab, Department of Chemistry Materials, and Chemical Engineering \u201cG. Natta\u201d, Politecnico di Milano Milan Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3735-2432","authenticated-orcid":false,"given":"Daniela Peneda","family":"Pacheco","sequence":"additional","affiliation":[{"name":"Bac<sup>3<\/sup>Gel\u00ae Lda, Edif\u00edcio Inova\u00e7\u00e3o II Porto Salvo Portugal"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1181-3632","authenticated-orcid":false,"given":"Livia","family":"Visai","sequence":"additional","affiliation":[{"name":"Molecular Medicine Department (DMM), Centre for Health Technologies (CHT), Unit\u00e0 di Ricerca (UdR) INSTM, Operative Unit (OU) of the Interuniversity Center for the Promotion of the 3Rs Principles in Teaching and Research (Centro 3R) University of Pavia Pavia Italy"},{"name":"Department of Prevention and Rehabilitation in Occupational Medicine and Speciality Medicine, UOR6 Nanotechnology Laboratory Istituti Clinici Scientifici Maugeri IRCCS Pavia Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2026,8,12]]},"reference":[{"key":"e_1_2_12_2_1","doi-asserted-by":"publisher","DOI":"10.1017\/S0007114512004023"},{"key":"e_1_2_12_3_1","doi-asserted-by":"publisher","DOI":"10.1086\/595011"},{"key":"e_1_2_12_4_1","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.01009-09"},{"key":"e_1_2_12_5_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.addr.2008.09.008"},{"key":"e_1_2_12_6_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.tim.2019.05.003"},{"key":"e_1_2_12_7_1","doi-asserted-by":"publisher","DOI":"10.1128\/mmbr.61.3.377-392.1997"},{"key":"e_1_2_12_8_1","doi-asserted-by":"publisher","DOI":"10.1128\/microbiolspec.bai-0013-2019"},{"key":"e_1_2_12_9_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41579-019-0158-9"},{"key":"e_1_2_12_10_1","doi-asserted-by":"publisher","DOI":"10.3390\/antibiotics15060537"},{"key":"e_1_2_12_11_1","doi-asserted-by":"publisher","DOI":"10.1093\/femsre\/fux010"},{"key":"e_1_2_12_12_1","doi-asserted-by":"publisher","DOI":"10.1099\/mic.0.000352"},{"key":"e_1_2_12_13_1","doi-asserted-by":"publisher","DOI":"10.1093\/ismejo\/wrae036"},{"key":"e_1_2_12_14_1","doi-asserted-by":"publisher","DOI":"10.1086\/314056"},{"key":"e_1_2_12_15_1","doi-asserted-by":"publisher","DOI":"10.3390\/microorganisms9050925"},{"key":"e_1_2_12_16_1","doi-asserted-by":"publisher","DOI":"10.1002\/bit.10376"},{"key":"e_1_2_12_17_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.tcb.2011.06.002"},{"key":"e_1_2_12_18_1","doi-asserted-by":"publisher","DOI":"10.3390\/cells11223634"},{"key":"e_1_2_12_19_1","doi-asserted-by":"publisher","DOI":"10.1128\/aem.01269-25"},{"key":"e_1_2_12_20_1","doi-asserted-by":"publisher","DOI":"10.3389\/fmicb.2019.00539"},{"key":"e_1_2_12_21_1","doi-asserted-by":"publisher","DOI":"10.1128\/microbiolspec.VMBF-0016-2015"},{"key":"e_1_2_12_22_1","volume-title":"Biosensors","author":"Nasiri R.","year":"2024"},{"key":"e_1_2_12_23_1","doi-asserted-by":"publisher","DOI":"10.3791\/2437"},{"key":"e_1_2_12_24_1","doi-asserted-by":"publisher","DOI":"10.1017\/S0950268812000465"},{"key":"e_1_2_12_25_1","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.01336-16"},{"key":"e_1_2_12_26_1","doi-asserted-by":"crossref","unstructured":"Pacheco D. 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