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These three\u2010dimensional bacterial aggregates are embedded in a self\u2010produced extracellular matrix that restricts antibiotic penetration and shields bacilli from host immune responses. The resulting spatial and physiological heterogeneity within biofilms generates microenvironments that favor slow\u2010growing or non\u2010replicating cells, markedly reducing the efficacy of conventional antimicrobial therapies. Increasing experimental and clinical evidence supports the presence of biofilm\u2010like mycobacterial communities in TB lesions, linking this growth mode to disease chronicity, treatment failure, and relapse. This review aims to provide an integrated overview of the biological and physiological states adopted by mycobacteria within biofilm microenvironments, with particular emphasis on the mechanisms underlying biofilm\u2010associated drug tolerance. In addition, it critically discusses therapeutic strategies designed to overcome this tolerance, focusing on synergistic antibiotic combinations and peptide\u2013antibiotic therapies that directly disrupt biofilm architecture, enhance drug penetration, or sensitize biofilm\u2010embedded bacilli to antimicrobial killing.\n                  <\/jats:p>","DOI":"10.1111\/apm.70166","type":"journal-article","created":{"date-parts":[[2026,3,10]],"date-time":"2026-03-10T16:19:18Z","timestamp":1773159558000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Biofilm Associated Persistence and Drug Tolerance in Mycobacteria Within Host Microenvironments"],"prefix":"10.1111","volume":"134","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-1830-2678","authenticated-orcid":false,"given":"Lourdes Serrano","family":"Garcia","sequence":"first","affiliation":[{"name":"School of Pharmaceutical Sciences S\u00e3o Paulo State University 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