{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,7]],"date-time":"2026-08-07T14:27:18Z","timestamp":1786112838950,"version":"build-2736575974"},"reference-count":56,"publisher":"Wiley","issue":"7","license":[{"start":{"date-parts":[[2026,7,8]],"date-time":"2026-07-08T00:00:00Z","timestamp":1783468800000},"content-version":"vor","delay-in-days":7,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"},{"start":{"date-parts":[[2026,7,1]],"date-time":"2026-07-01T00:00:00Z","timestamp":1782864000000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["J Biomed Mater Res"],"published-print":{"date-parts":[[2026,7]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:p>\n                    This study compared four biofilm fixation protocols for scanning electron microscopy (SEM) imaging and their compatibility with Fiji\u2010based image analysis, and introduced a complementary approach for SEM biofilm cross\u2010section analysis to be used when assessing the antimicrobial activity of dental restorative materials. In the first phase, resin composite discs (8\u2009\u00d7\u20092\u2009mm) were characterized for degree of conversion and surface roughness, then incubated with renilla\u2010reporter\n                    <jats:styled-content style=\"fixed-case\">\n                      <jats:italic>Streptococcus mutans<\/jats:italic>\n                    <\/jats:styled-content>\n                    for 1 and 24\u2009h. Metabolic activity was assessed by luciferase assay, and biofilms were fixed using four protocols based on buffered formalin and\/or glutaraldehyde, with or without graded ethanol dehydration, prior to SEM imaging. In parallel, biofilm cross\u2010sections were analyzed using resin bars incubated under different conditions. In the second phase, the most effective fixation protocol was validated using antimicrobial\u2010containing restorative materials, including two resin composites and a glass ionomer cement. Data were analyzed using ANOVA and Tukey's test (\n                    <jats:italic>p<\/jats:italic>\n                    \u2009&lt;\u20090.05). The results demonstrated that biofilm fixation protocols significantly influence SEM image quality, biofilm architectural features, and quantitative measurements obtained through image analysis. In conclusion, standardized biofilm fixation using glutaraldehyde combined with graded ethanol dehydration and coupled with machine\u2010learning augmented analysis of SEM images is essential for producing superior image resolution to enable clearer discrimination among bacterial cells, biofilm matrix, and material surfaces, and improved sensitivity in detecting architectural differences, particularly in biofilm cross\u2010sections.\n                  <\/jats:p>","DOI":"10.1002\/jbm.b.70125","type":"journal-article","created":{"date-parts":[[2026,7,8]],"date-time":"2026-07-08T07:42:11Z","timestamp":1783496531000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Quantitative and Structural Characterization of\n                    <scp>\n                      <i>Streptococcus mutans<\/i>\n                    <\/scp>\n                    Biofilms on Dental Restorative Materials Using Scanning Electron Microscopy and Machine Learning"],"prefix":"10.1002","volume":"114","author":[{"given":"Krishna Kumar","family":"Kungumaraj","sequence":"first","affiliation":[{"name":"Department of Biomaterial and Biomedical Sciences Oregon Health &amp; Science University \u2010 School of Dentistry  Portland Oregon USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sivashankari","family":"Rajasekaran","sequence":"additional","affiliation":[{"name":"Department of Biomaterial and Biomedical Sciences Oregon Health &amp; Science University \u2010 School of Dentistry  Portland Oregon USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bao","family":"Huynh","sequence":"additional","affiliation":[{"name":"Department of Biomaterial and Biomedical Sciences Oregon Health &amp; Science University \u2010 School of Dentistry  Portland Oregon 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