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This study aimed to evaluate the effects of enzymatic-based CMCR on the proliferation and osteogenic differentiation of dental pulp stem cells (DPSCs).<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>MTT assay, scratch assay, and gene expression analysis were conducted to assess cell proliferation (viability percentage), migration (wound healing percentage), and osteogenic differentiation (RT-qPCR). The viability of DPSCs after direct exposure to enzymatic-based CMCR (0.5%, 1%, and 2% in DMEM) was measured at 24 and 48\u00a0h post-exposure. Indirect exposure was also tested using sound mineralized (0.5\u00a0mm and 1\u00a0mm) and demineralized (0.5\u00a0mm) dentin discs. Scratch assay was performed for control, direct, and indirect groups at 0, 24, and 48\u00a0h, followed by fixation and crystal violet staining at 48\u00a0h. Dentin sialophosphoprotein (DSPP) gene expression was quantified using RT-qPCR after 48\u00a0h.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>\n                      Two-way ANOVA of proliferation and migration assays revealed that both indirect (0.5\u00a0mm demineralized dentin) and direct (0.5% DMEM) exposure groups showed the lowest cytotoxic effect after 24\u00a0h (82.75\u2009\u00b1\u20093.37 and 68.11\u2009\u00b1\u20094.99, respectively). After 48\u00a0h, a recovery trend in DPSC viability was observed, with both groups exhibiting the highest proliferation (93.60\u2009\u00b1\u20094.67 and 78.25\u2009\u00b1\u20094.88, respectively). These groups also showed the highest wound closure percentages (78.78\u2009\u00b1\u20090.89 for direct and 74.19\u2009\u00b1\u20091.03 for indirect exposure). One-way ANOVA of RT-qPCR results demonstrated significant upregulation of DSPP gene expression in the indirect (0.5\u00a0mm demineralized dentin) group (5.35\u2009\u00b1\u20092.01,\n                      <jats:italic>p<\/jats:italic>\n                      \u2009&lt;\u20090.05).\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusion<\/jats:title>\n                    <jats:p>Enzymatic-based CMCR exhibits favorable biocompatibility, particularly when applied directly on DPSCs at low concentrations and when used indirectly through dentin discs. These findings suggest that this enzymatic caries removal approach is potentially safe, effective, and minimally invasive, making it suitable for conservative restorative dentistry.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.1186\/s12903-025-07495-w","type":"journal-article","created":{"date-parts":[[2025,12,27]],"date-time":"2025-12-27T02:23:05Z","timestamp":1766802185000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Effect of Enzymatic-based chemomechanical caries removal agent on proliferation and osteogenic differentiation of dental pulp stem cells"],"prefix":"10.1186","volume":"26","author":[{"given":"Hend A.","family":"Gouda","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hamdi H.","family":"Hamama","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Menatalla M.","family":"Elhindawy","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Asmaa S.","family":"Elmahdy","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Youssry M.","family":"Elhawary","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"297","published-online":{"date-parts":[[2025,12,27]]},"reference":[{"issue":"Suppl 1Suppl 1","key":"7495_CR1","doi-asserted-by":"publisher","DOI":"10.1186\/1472-6831-6-S1-S2","volume":"6","author":"BL Edelstein","year":"2006","unstructured":"Edelstein BL. 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The included premolars in this study underwent extraction for therapeutic indications not linked to the study. Informed consents were taken from all persons receiving dental care at the Oral and Maxillofacial Surgery Department Clinic, Faculty of Dentistry, Mansoura University. They donated their extracted premolars voluntarily to the researchers to conduct the study. The ethics committee of Faculty of Dentistry, Mansoura University granted ethical approval (No.A02020822) to this study prior to commencement.The authors declare no conflicts of interest. The authors alone are responsible for the content and writing of this paper","order":2,"name":"Ethics","group":{"name":"EthicsHeading","label":"Ethics approval and consent for publication"}},{"value":"The authors declare no competing interests.","order":3,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing interests"}},{"value":"Not applicable.","order":4,"name":"Ethics","group":{"name":"EthicsHeading","label":"Consent for publication"}}],"article-number":"182"}}