{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,12,15]],"date-time":"2025-12-15T14:21:24Z","timestamp":1765808484658,"version":"build-2065373602"},"reference-count":49,"publisher":"MDPI AG","issue":"9","license":[{"start":{"date-parts":[[2025,9,10]],"date-time":"2025-09-10T00:00:00Z","timestamp":1757462400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Coordination for the Improvement of Higher Education Personnel, Brazil","award":["001","2021\/02260-6","23\/2025"],"award-info":[{"award-number":["001","2021\/02260-6","23\/2025"]}]},{"name":"S\u00e3o Paulo Research Foundation","award":["001","2021\/02260-6","23\/2025"],"award-info":[{"award-number":["001","2021\/02260-6","23\/2025"]}]},{"name":"PROPG\/UNESP","award":["001","2021\/02260-6","23\/2025"],"award-info":[{"award-number":["001","2021\/02260-6","23\/2025"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Dentistry Journal"],"abstract":"<jats:p>Objectives: This study aimed to evaluate whether the addition of pressurized carbon dioxide (PCD) influences the antimicrobial efficacy of 2.5% sodium hypochlorite (NaOCl) against Enterococcus faecalis biofilm in root canals and dentinal tubules. Methods: Forty extracted human mandibular premolars with single canals were contaminated with E. faecalis for 10 days and randomly assigned to four groups (n = 10): 2.5% NaOCl, 2.5% NaOCl + CO2, sterile saline, and sterile saline + CO2. The pH and temperature of the NaOCl solution were measured before and after CO2 incorporation. Microbial load was assessed by CFU counts before and after irrigation, and in dentin samples from the cervical, middle, and apical thirds. Oxidative stress was evaluated via lipid peroxidation (TBARS), protein carbonyl content, and total protein quantification. Biofilm metabolic activity was analyzed using the XTT reduction assay. Data were analyzed using one-way ANOVA on ranks and two-way repeated measures ANOVA (\u03b1 = 0.05), a very large effect size (Cohen\u2019s d) \u2248 1.756 was assumed. Results: All irrigation protocols significantly reduced bacterial load (p &lt; 0.05). Both NaOCl groups outperformed the saline controls (p = 0.009). The addition of CO2 to NaOCl slightly enhanced disinfection in the main canal but did not improve antimicrobial action in dentinal tubules. CO2 incorporation reduced the pH of NaOCl from ~13.4 to 7.4 and slightly increased the temperature, making the solution more chemically reactive. However, both oxidative stress markers and the XTT assay showed that the combination with CO2 impaired the antimicrobial effectiveness of NaOCl. Conclusions: Despite the improvement in bacterial reduction in the root canal lumen, the combination of PCD with NaOCl failed to enhance intratubular disinfection and reduced the oxidative damage and metabolic inactivation of the biofilm. CO2 pressurization appears to limit the antimicrobial action of NaOCl.<\/jats:p>","DOI":"10.3390\/dj13090417","type":"journal-article","created":{"date-parts":[[2025,9,10]],"date-time":"2025-09-10T08:43:54Z","timestamp":1757493834000},"page":"417","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Synergistic Effect of Sodium Hypochlorite and Carbon Dioxide Against Enterococcus faecalis Biofilm"],"prefix":"10.3390","volume":"13","author":[{"given":"J\u00falia Guerra","family":"de Andrade","sequence":"first","affiliation":[{"name":"Department of Restorative Dentistry, Ara\u00e7atuba School of Dentistry, S\u00e3o Paulo State University (UNESP), Ara\u00e7atuba 16015-050, SP, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1948-862X","authenticated-orcid":false,"given":"Ana Fl\u00e1via Folhas","family":"Natali","sequence":"additional","affiliation":[{"name":"Department of Restorative Dentistry, Ara\u00e7atuba School of Dentistry, S\u00e3o Paulo State University (UNESP), Ara\u00e7atuba 16015-050, SP, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Caroline","family":"Loureiro","sequence":"additional","affiliation":[{"name":"Department of Restorative Dentistry, Ara\u00e7atuba School of Dentistry, S\u00e3o Paulo State University (UNESP), Ara\u00e7atuba 16015-050, SP, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Gladiston William Lobo","family":"Rodrigues","sequence":"additional","affiliation":[{"name":"Department of Restorative Dentistry, Ara\u00e7atuba School of Dentistry, S\u00e3o Paulo State University (UNESP), Ara\u00e7atuba 16015-050, SP, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ana Paula Fernandes","family":"Ribeiro","sequence":"additional","affiliation":[{"name":"Department of Restorative Dentistry, Ara\u00e7atuba School of Dentistry, S\u00e3o Paulo State University (UNESP), Ara\u00e7atuba 16015-050, SP, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Rayara Nogueira","family":"de Freitas","sequence":"additional","affiliation":[{"name":"Department of Basic Sciences, Ara\u00e7atuba School of Dentistry, S\u00e3o Paulo State University (UNESP), Ara\u00e7atuba 16015-050, SP, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Renan Jose","family":"Barzotti","sequence":"additional","affiliation":[{"name":"Department of Basic Sciences, Ara\u00e7atuba School of Dentistry, S\u00e3o Paulo State University (UNESP), Ara\u00e7atuba 16015-050, SP, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Laura Ces\u00e1rio","family":"Oliveira","sequence":"additional","affiliation":[{"name":"Department of Restorative Dentistry, Ara\u00e7atuba School of Dentistry, S\u00e3o Paulo State University (UNESP), Ara\u00e7atuba 16015-050, SP, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0009-0003-6111-5311","authenticated-orcid":false,"given":"Yuri Gabriel Chamorro","family":"de Moraes","sequence":"additional","affiliation":[{"name":"Department of Restorative Dentistry, Ara\u00e7atuba School of Dentistry, S\u00e3o Paulo State University (UNESP), Ara\u00e7atuba 16015-050, SP, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Nat\u00e1lia Amanda","family":"Gomes","sequence":"additional","affiliation":[{"name":"Department of Restorative Dentistry, Ara\u00e7atuba School of Dentistry, S\u00e3o Paulo State University (UNESP), Ara\u00e7atuba 16015-050, SP, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6481-5506","authenticated-orcid":false,"given":"Ant\u00f4nio Hernandes","family":"Chaves-Neto","sequence":"additional","affiliation":[{"name":"Department of Basic Sciences, Ara\u00e7atuba School of Dentistry, S\u00e3o Paulo State University (UNESP), Ara\u00e7atuba 16015-050, SP, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4140-340X","authenticated-orcid":false,"given":"Frederico Canato","family":"Martinho","sequence":"additional","affiliation":[{"name":"Department of Advanced Oral Sciences and Therapeutics, School of Dentistry, University of Maryland, Baltimore, MD 21201, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2362-8920","authenticated-orcid":false,"given":"Rog\u00e9rio de Castilho","family":"Jacinto","sequence":"additional","affiliation":[{"name":"Department of Restorative Dentistry, Ara\u00e7atuba School of Dentistry, S\u00e3o Paulo State University (UNESP), Ara\u00e7atuba 16015-050, SP, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2025,9,10]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"539","DOI":"10.1016\/j.sdentj.2024.01.009","article-title":"Prevalence of Enterococcus faecalis and Candida albicans in endodontic retreatment Cases: A comprehensive study","volume":"36","author":"Ahmed","year":"2024","journal-title":"Saudi Dent. 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