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However, these antimicrobials can lead to microbial resistance and chlorhexidine can impair health-associated nitrate (NO\n                    <jats:sub>3<\/jats:sub>\n                    <jats:sup>-<\/jats:sup>\n                    ) metabolism. We tested the effect of chlorhexidine (0.002%), metronidazole (16\u00a0\u00b5g\/ml) and amoxicillin (0.7\u00a0\u00b5g\/ml), with and without 8 mM NO\n                    <jats:sub>3<\/jats:sub>\n                    <jats:sup>-<\/jats:sup>\n                    , on the bacterial composition and NO\n                    <jats:sub>3<\/jats:sub>\n                    <jats:sup>-<\/jats:sup>\n                    metabolism of subgingival plaque samples from 12 periodontitis patients grown in vitro for 8\u00a0h. The low sublethal concentrations of amoxicillin and chlorhexidine significantly inhibited microbial growth and impaired NO\n                    <jats:sub>3<\/jats:sub>\n                    <jats:sup>-<\/jats:sup>\n                    reduction, whereas the physiological concentration of metronidazole did not. A lower subgingival microbial dysbiosis index (SMDI) was found in the NO\n                    <jats:sub>3<\/jats:sub>\n                    <jats:sup>-<\/jats:sup>\n                    condition compared with amoxicillin alone and chlorhexidine with or without NO\n                    <jats:sub>3<\/jats:sub>\n                    <jats:sup>-<\/jats:sup>\n                    (\n                    <jats:italic>p<\/jats:italic>\n                    \u2009&lt;\u20090.05). The SMDI of the metronidazole conditions was also significantly lower than in those with chlorhexidine (\n                    <jats:italic>p<\/jats:italic>\n                    \u2009&lt;\u20090.05). Moreover, NO\n                    <jats:sub>3<\/jats:sub>\n                    <jats:sup>-<\/jats:sup>\n                    alone or combined with metronidazole appeared to increase\n                    <jats:italic>Neisseria<\/jats:italic>\n                    spp. and\n                    <jats:italic>Aggregatibacter<\/jats:italic>\n                    spp., whilst disease-associated changes were found in the chlorhexidine and amoxicillin conditions. Adding NO\n                    <jats:sub>3<\/jats:sub>\n                    <jats:sup>-<\/jats:sup>\n                    to metronidazole led to health-associated changes compared with metronidazole alone. In conclusion, low levels of amoxicillin and chlorhexidine limited microbial growth, impaired NO\n                    <jats:sub>3<\/jats:sub>\n                    <jats:sup>-<\/jats:sup>\n                    metabolism and were linked to disease-associated microbial profiles. A dual treatment of metronidazole\u2009+\u2009NO\n                    <jats:sub>3<\/jats:sub>\n                    <jats:sup>-<\/jats:sup>\n                    should be further investigated in clinical studies.\n                  <\/jats:p>","DOI":"10.1038\/s41598-026-36588-x","type":"journal-article","created":{"date-parts":[[2026,1,30]],"date-time":"2026-01-30T07:04:45Z","timestamp":1769756685000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Combined effects of nitrate and antimicrobial compounds on in vitro subgingival biofilms"],"prefix":"10.1038","volume":"16","author":[{"given":"Siobhan P.","family":"Moran","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Maria","family":"Nadal-Ruiz","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alex","family":"Mira","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ana","family":"Serrano-Valcarce","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mercedes","family":"Mompe\u00e1n","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Migl\u0117","family":"\u017diemyt\u0117","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Andr\u00e9s","family":"L\u00f3pez-Rold\u00e1n","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fiona L.","family":"Henriquez","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mia C.","family":"Burleigh","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bob T.","family":"Rosier","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2026,1,30]]},"reference":[{"key":"36588_CR1","doi-asserted-by":"publisher","first-page":"462","DOI":"10.1111\/idj.12630","volume":"71","author":"T Kwon","year":"2021","unstructured":"Kwon, T., Lamster, I. 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