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However, worries about synthetic additives\u2019 safety and long-term effects urge the use of natural, biocompatible alternatives. This study explored the antibacterial efficacy of chitosan (CH) and nano-chitosan (NCH) incorporated into different restorative materials: amalgam (Am), glass ionomer cement (G), and resin composite (R). This study employs a direct comparative analysis of both modified and unmodified restorative materials against Am to evaluate their efficacy against\n                    <jats:italic>Streptococcus mutans<\/jats:italic>\n                    ATCC 25,175 using agar diffusion, broth inhibition, biofilm formation assays, and qPCR investigation of virulence genes (\n                    <jats:italic>ldh<\/jats:italic>\n                    and\n                    <jats:italic>gtfB<\/jats:italic>\n                    ). Incorporating CH and NCH, especially at 15%, decreased biofilm biomass, downregulated virulence gene expression, and significantly increased the effectiveness of bacterial growth suppression. When compared to controls, the G modified with 15% NCH (GN15) showed the strongest inhibition of bacterial growth by 43.1%. The most significant finding was the ability of GN15 to downregulate the expression of\n                    <jats:italic>ldh<\/jats:italic>\n                    and\n                    <jats:italic>gtfB<\/jats:italic>\n                    by 50% and 28%, respectively, indicating a disruption in acid production and biofilm matrix synthesis. While Am showed the highest overall inhibition (86%, 65%). These results demonstrate that nano-chitosan is a biocompatible and promising addition to restorative materials, providing improved defense against cariogenic biofilms and facilitating the creation of dental biomaterials of the future.\n                  <\/jats:p>","DOI":"10.1186\/s13568-025-02004-2","type":"journal-article","created":{"date-parts":[[2026,2,2]],"date-time":"2026-02-02T23:45:39Z","timestamp":1770075939000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Nano-chitosan modified restorative materials suppress Streptococcus mutans biofilm and virulence gene expression"],"prefix":"10.1186","volume":"16","author":[{"given":"Jakline N.","family":"Saddik","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Martha M.","family":"Naguib","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Labib M.","family":"Labib","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ahmed O.","family":"El-Gendy","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fatma","family":"Molham","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2026,2,3]]},"reference":[{"key":"2004_CR1","doi-asserted-by":"crossref","unstructured":"Amin FR, Khurshid S, Zafar Z, Sefat MS, Kumar F, N. 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