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This study evaluated bacterial nanocellulose (BNC) as a scaffold for the 3D culture of colorectal cancer cells. BNC scaffolds were produced using paraffin spheres as porogens and subsequently characterized in terms of morphology, chemical structure, thermal stability, and mechanical properties. Human colorectal cancer cell lines SW480 and SW620, along with CHO\u2010K1 cells, were exposed to BNC leachates and cultured on 2D and 3D BNC matrices. Cells grown on 2D BNC exhibited typical monolayer proliferation, whereas 3D matrices allowed cell infiltration and volumetric distribution, supporting multicellular organization and the formation of spheroid\u2010like aggregates. Cell viability remained in both leachate and direct scaffold cultures. Genotoxicity assays showed low levels of DNA damage, with most cells displaying no detectable genotoxic effects, and mutagenicity tests revealed no significant differences compared to controls (p &gt; 0.05). Overall, the results demonstrate that 3D BNC supports cell viability, multicellular organization, and genomic safety, highlighting its suitability as a biocompatible scaffold for in vitro 3D colon cancer models.<\/jats:p>","DOI":"10.1002\/mabi.70272","type":"journal-article","created":{"date-parts":[[2026,10,4]],"date-time":"2026-10-04T17:00:11Z","timestamp":1791133211000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Bacterial Nanocellulose as a Platform for Three\u2010Dimensional Colon Cancer Cultures: Physicochemical Characterization and Biocompatibility Evaluation"],"prefix":"10.1002","volume":"26","author":[{"given":"M.","family":"Londo\u00f1o","sequence":"first","affiliation":[{"name":"Systems Biology Research Group School of Health Sciences Faculty of Medicine Universidad Pontificia Bolivariana  Medell\u00edn Colombia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"C.","family":"Castro","sequence":"additional","affiliation":[{"name":"New Materials Research Group, School of Engineering Universidad Pontificia Bolivariana Medell\u00edn Colombia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"A.","family":"Ca\u00f1as","sequence":"additional","affiliation":[{"name":"New Materials Research Group, School of Engineering Universidad Pontificia Bolivariana Medell\u00edn Colombia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"S.","family":"Ruiz","sequence":"additional","affiliation":[{"name":"NanoBioCel Group Department of Pharmacy and Food Science Faculty of Pharmacy University of the Basque Country (UPV\/ EHU)  Vitoria\u2010Gasteiz Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"J.","family":"Pedraz","sequence":"additional","affiliation":[{"name":"NanoBioCel Group Department of Pharmacy and Food Science Faculty of Pharmacy University of the Basque Country (UPV\/ EHU)  Vitoria\u2010Gasteiz Spain"},{"name":"Joint Research Laboratory (JRL) on Advanced Pharma Development Centro de investigac\u00f3n Lascaray ikergunea A Joint Venture of TECNALIA and University of the Basque Country  Vitoria\u2010Gasteiz Spain"},{"name":"Centro de Investigaci\u00f3n Biom\u00e9dica en Red de Bioingenier\u00eda Biomateriales y Nanomedicina (CIBER\u2010BBN) Instituto De Salud Carlos III  Vitoria\u2010Gasteiz Spain"},{"name":"Bioaraba NanoBioCel Research Group  Vitoria\u2010Gasteiz Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7755-5846","authenticated-orcid":false,"given":"M.","family":"Osorio","sequence":"additional","affiliation":[{"name":"Systems Biology Research Group School of Health Sciences Faculty of Medicine Universidad Pontificia Bolivariana  Medell\u00edn Colombia"},{"name":"New Materials Research Group, School of Engineering Universidad Pontificia Bolivariana Medell\u00edn Colombia"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2026,10,4]]},"reference":[{"key":"e_1_2_10_2_1","doi-asserted-by":"publisher","DOI":"10.3390\/cancers14071732"},{"key":"e_1_2_10_3_1","doi-asserted-by":"publisher","DOI":"10.3322\/CAAC.21660"},{"key":"e_1_2_10_4_1","doi-asserted-by":"publisher","DOI":"10.1001\/jama.2021.0106"},{"key":"e_1_2_10_5_1","doi-asserted-by":"publisher","DOI":"10.3389\/fmed.2022.995882"},{"key":"e_1_2_10_6_1","doi-asserted-by":"publisher","DOI":"10.4155\/bio.12.133"},{"key":"e_1_2_10_7_1","doi-asserted-by":"publisher","DOI":"10.5114\/aoms.2016.63743"},{"key":"e_1_2_10_8_1","doi-asserted-by":"crossref","first-page":"45","DOI":"10.1016\/B978-0-12-804667-8.00003-1","volume-title":"In Vitro Toxicology","author":"Jain A. 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