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Lyophilized CFS of each probiotic strain was individually mixed with Poloxamer gel using cold technique to prepare probiotic supernatants (PS) gel. Dentin blocks from cervical root portions were standardized at similar dimensions, infected with\n                      <jats:italic>E. faecalis<\/jats:italic>\n                      for 21 days to generate a mature biofilm. At the end of incubation, infected dentin blocks were randomly assigned to five groups (\n                      <jats:italic>n<\/jats:italic>\n                      \u2009=\u200910) based on the medication as follows: positive control (PC), calcium hydroxide (CH),\n                      <jats:italic>Lactobacillus acidophilus<\/jats:italic>\n                      PS gel (LA),\n                      <jats:italic>Lactobacillus bulgaricus<\/jats:italic>\n                      PS gel (LB),\n                      <jats:italic>Lactobacillus<\/jats:italic>\n                      combination PS gel (LC) combining LA and LB at a 1:1 ratio. All dentin blocks were medicated and incubated for 7 days under anaerobic conditions. Then, dentin blocks were examined with CLSM at 40X magnification; three-dimensional imaging in Z-stack mode was processed, and the dead\n                      <jats:italic>E. faecalis<\/jats:italic>\n                      % was quantitatively assessed using bioImage_L v21 software and total biovolume was calculated. Dead\n                      <jats:italic>E. faecalis<\/jats:italic>\n                      % were compared across groups using a one-way ANOVA test followed by Tukey\u2019s post hoc test. Biovolume data were compared using the Kruskal-Wallis test followed by Dunn\u2019s post hoc test with p-value adjustment using Holm\u2019s method. The significance level was set at\n                      <jats:italic>p<\/jats:italic>\n                      \u2009&lt;\u20090.05 for all tests. Statistical analysis was performed using R version 4.6.0 for Windows.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>\n                      LB was linked to the highest percentage of dead\n                      <jats:italic>E. faecalis<\/jats:italic>\n                      % (\n                      <jats:italic>p<\/jats:italic>\n                      \u2009&lt;\u20090.05), followed by LC, LA, CH, and the PC. Significant differences in dead\n                      <jats:italic>E. faecalis<\/jats:italic>\n                      % were observed between all groups (\n                      <jats:italic>p<\/jats:italic>\n                      \u2009&lt;\u20090.05) except for CH and the PC groups. There was no significant difference in total biovolume among all groups.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusion<\/jats:title>\n                    <jats:p>The Initial assessment of LB showed a promising antimicrobial action in dentin model that exceeded LA and CH. As biofilm biovolume was unaffected, LB appears to reduce bacterial viability rather than remove biofilm mass, warranting further validation in polymicrobial and intact canal models.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Clinical significance<\/jats:title>\n                    <jats:p>\n                      Probiotics may be of clinical relevance due to their antimicrobial action against\n                      <jats:italic>E. faecalis<\/jats:italic>\n                      .\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.1007\/s00784-026-07125-x","type":"journal-article","created":{"date-parts":[[2026,9,7]],"date-time":"2026-09-07T02:26:25Z","timestamp":1788747985000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["A preliminary antimicrobial investigation of lactobacillus bulgaricus probiotic supernatants as an intracanal medication against mature E. faecalis biofilm in an ex-vivo dentin blocks model"],"prefix":"10.1007","volume":"30","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-7198-9595","authenticated-orcid":false,"given":"Sherif","family":"Heidar","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4111-3541","authenticated-orcid":false,"given":"Hayam Y.","family":"Hassan","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4443-2345","authenticated-orcid":false,"given":"Edgar","family":"Sch\u00e4fer","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1465-3198","authenticated-orcid":false,"given":"Shehabeldin","family":"Saber","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2026,9,7]]},"reference":[{"key":"7125_CR1","doi-asserted-by":"publisher","first-page":"509","DOI":"10.1016\/j.joen.2015.12.014","volume":"42","author":"D Ricucci","year":"2016","unstructured":"Ricucci D, Candeiro GT, Bugea C, Siqueira JF (2016) Jr. Complex Apical Intraradicular Infection and Extraradicular Mineralized Biofilms as the Cause of Wet Canals and Treatment Failure: Report of 2 Cases. 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