{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,24]],"date-time":"2025-10-24T05:34:05Z","timestamp":1761284045296,"version":"build-2065373602"},"reference-count":29,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2025,10,24]],"date-time":"2025-10-24T00:00:00Z","timestamp":1761264000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Dent. Med."],"abstract":"<jats:sec>\n                    <jats:title>Objectives<\/jats:title>\n                    <jats:p>Dental pulp tissue presents unique challenges for RNA analysis due to its fibrous nature, elevated RNase expression, and susceptibility to degradation during extraction procedures. This study systematically evaluated three distinct preservation methodologies to determine optimal approaches for maintaining RNA integrity and quality in human dental pulp tissue.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>Dental pulp samples were obtained from thirty-six patients diagnosed with irreversible pulpitis requiring endodontic treatment. Tissues were preserved using three methods: snap freezing in liquid nitrogen, RNAiso Plus reagent preservation, and RNAlater solution storage. RNA quality was comprehensively assessed using multiple complementary approaches; Nanodrop spectrophotometry, Qubit fluorometry, and Bioanalyzer capillary electrophoresis. Parameters assessed were yield quantification, purity assessment and structural integrity.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>\n                      RNAlater storage demonstrated statistically significant superior performance across all evaluated parameters. Yield analysis showed an 11.5-fold enhancement relative to snap freezing (4,425.92\u2009\u00b1\u20092,299.78 vs. 384.25\u2009\u00b1\u2009160.82\u2005ng\/\u03bcl,\n                      <jats:italic>p<\/jats:italic>\n                      \u2009&amp;lt;\u20090.001) and 1.8-fold improvement over RNAiso Plus extraction. Integrity assessment indicated significant advantages with RNAlater samples exhibiting mean RIN values of 6.0\u2009\u00b1\u20092.07 compared to snap freezing at 3.34\u2009\u00b1\u20092.87 (\n                      <jats:italic>p<\/jats:italic>\n                      \u2009=\u20090.028). Quality evaluation demonstrated RNAlater samples achieved optimal RNA quality in 75% of cases while snap freezing achieved this standard in only 33% of samples.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusion<\/jats:title>\n                    <jats:p>RNAlater storage establishes as the optimal preservation approach for dental pulp RNA investigations, delivering enhanced yield, purity, and integrity parameters. The results furnish credible affirmation for establishing methodological standardisation in dental transcriptomics investigations and clinical implementations.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.3389\/fdmed.2025.1659932","type":"journal-article","created":{"date-parts":[[2025,10,24]],"date-time":"2025-10-24T05:28:15Z","timestamp":1761283695000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":0,"title":["RNA preservation in human dental pulp for transcriptomic profiling: a comparative multi-parameter study"],"prefix":"10.3389","volume":"6","author":[{"given":"Raksha","family":"Bhat","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Preethesh","family":"Shetty","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Shishir","family":"Shetty","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1965","published-online":{"date-parts":[[2025,10,24]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","first-page":"647","DOI":"10.3892\/ijmm.2017.3065","article-title":"Comparison of gene expression profiles between dental pulp and periodontal ligament tissues in humans","volume":"40","author":"Gong","year":"2017","journal-title":"Int J Mol Med"},{"key":"B2","doi-asserted-by":"publisher","first-page":"105646","DOI":"10.1016\/j.archoralbio.2023.105646","article-title":"Comparison of human dental tissue RNA extraction methods for RNA sequencing","volume":"148","author":"Zhao","year":"2023","journal-title":"Arch Oral Biol"},{"key":"B3","doi-asserted-by":"publisher","first-page":"e18072","DOI":"10.7717\/peerj.18072","article-title":"Optimizing total RNA extraction method for human and mice samples","volume":"12","author":"Zeng","year":"2024","journal-title":"PeerJ"},{"key":"B4","doi-asserted-by":"publisher","first-page":"116","DOI":"10.1038\/modpathol.3880267","article-title":"Preservation of RNA for functional genomic studies: a multidisciplinary tumor bank protocol","volume":"14","author":"Florell","year":"2001","journal-title":"Mod Pathol"},{"key":"B5","doi-asserted-by":"publisher","first-page":"88","DOI":"10.1186\/1471-2164-5-88","article-title":"Comparison of frozen and RNALater solid tissue storage methods for use in RNA expression microarrays","volume":"5","author":"Ramos","year":"2004","journal-title":"BMC Genomics"},{"key":"B6","doi-asserted-by":"publisher","first-page":"5303","DOI":"10.3390\/ma13225303","article-title":"Advanced biomaterials and techniques for oral tissue engineering and regeneration-A review","volume":"13","author":"Matichescu","year":"2020","journal-title":"Materials (Basel)"},{"key":"B7","doi-asserted-by":"publisher","first-page":"807","DOI":"10.1136\/jclinpath-2012-201283","article-title":"Preservation of nucleic acids and tissue morphology in paraffin-embedded clinical samples: comparison of five molecular fixatives","volume":"66","author":"Staff","year":"2013","journal-title":"J Clin Pathol"},{"year":"2024","key":"B8","article-title":"Preserving RNA integrity in tissue samples prior to RNA isolation. 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