{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,23]],"date-time":"2026-09-23T10:07:18Z","timestamp":1790158038469,"version":"4.1.0"},"reference-count":49,"publisher":"Wiley","issue":"9","license":[{"start":{"date-parts":[[2026,9,23]],"date-time":"2026-09-23T00:00:00Z","timestamp":1790121600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2026,9,23]],"date-time":"2026-09-23T00:00:00Z","timestamp":1790121600000},"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":["European Journal of Pain"],"published-print":{"date-parts":[[2026,10]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:sec>\n                    <jats:title>Background<\/jats:title>\n                    <jats:p>Accurate evaluation of pain\u2010related behaviours in rodent models is crucial for elucidating pain mechanisms and assessing the efficacy of novel analgesic compounds. Traditional nociceptive assays primarily measure evoked responses, which have limited translational relevance since spontaneous pain is the predominant symptom in pain patients. Rodent models of spontaneous pain reveal innate behaviours, including paw licking and flinching, but their manual quantification is often subject to observer bias, is error\u2010prone and labor\u2010intensive.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>We have developed a deep\u2010learning\u2010based algorithm designed to automatically detect and quantify spontaneous pain behaviours in mice from standard 2D bottom\u2010up video recordings. Focusing on paw licking and flinching in the capsaicin pain model, and validated in the formalin test and a model of cancer\u2010evoked pain, our approach provides an objective, high\u2010throughput and reproducible alternative to traditional manual scoring.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>\n                      The pipeline, combining DeepLabCut\u2010based keypoint tracking with a transformer classifier, achieved 92% balanced accuracy in identifying licking, flinching and baseline behaviours in the capsaicin model. Automated quantification showed an excellent correlation with expert human annotations (Pearson's\n                      <jats:italic>r<\/jats:italic>\n                      \u2009=\u20090.99), while frame\u2010by\u2010frame comparisons with blinded observers showed high agreement and minimal bias. Performance was maintained on an independently generated external dataset (Pearson's\n                      <jats:italic>r<\/jats:italic>\n                      \u2009=\u20090.98), supporting reproducibility across experimental settings. Validation in inflammatory and cancer pain models further demonstrated robust generalizability across distinct pathological conditions.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusion<\/jats:title>\n                    <jats:p>By reducing observer\u2010dependent variability and capturing subtle temporal and spatial dynamics, this framework provides a standardized and accessible tool for the objective assessment of ongoing nociceptive responses, facilitating comparisons across distinct experimental models and institutional settings.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Significance Statement<\/jats:title>\n                    <jats:p>Our findings highlight the potential of deep learning to advance preclinical pain research by enabling standardized, unbiased and clinically relevant assessment of spontaneous pain in rodent models.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.1002\/ejp.70391","type":"journal-article","created":{"date-parts":[[2026,9,23]],"date-time":"2026-09-23T09:28:00Z","timestamp":1790155680000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Deep Learning\u2010Based Quantification of Spontaneous Pain Behaviours in Mice"],"prefix":"10.1002","volume":"30","author":[{"given":"Michele","family":"Zanoletti","sequence":"first","affiliation":[{"name":"Institute of Clinical Physiology National Research Council  Pisa Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Elisa","family":"Bellantoni","sequence":"additional","affiliation":[{"name":"Department of Health Sciences Section of Clinical Pharmacology and Oncology University of Florence  Florence Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"L\u00eddia","family":"Pombo Gomes","sequence":"additional","affiliation":[{"name":"Graduate Program of Neurosciences, Institute of Biology Fluminense Federal University  Rio de Janeiro Niteroi Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Maria Luiza","family":"Quadros dos Santos","sequence":"additional","affiliation":[{"name":"Graduate Program of Neurosciences, Institute of Biology Fluminense Federal University  Rio de Janeiro Niteroi Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Daniel","family":"Souza Monteiro de Ara\u00fajo","sequence":"additional","affiliation":[{"name":"Graduate Program of Neurosciences, Institute of Biology Fluminense Federal University  Rio de Janeiro Niteroi Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gaetano","family":"De Siena","sequence":"additional","affiliation":[{"name":"Department of Health Sciences Section of Clinical Pharmacology and Oncology University of Florence  Florence Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sabrina","family":"Qader Kudsi","sequence":"additional","affiliation":[{"name":"Graduate Program in Pharmacology Federal University of Santa Maria (UFSM)  Santa Maria Rio Grande do Sul Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9223-8395","authenticated-orcid":false,"given":"Romina","family":"Nassini","sequence":"additional","affiliation":[{"name":"Department of Health Sciences Section of Clinical Pharmacology and Oncology University of Florence  Florence Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marco","family":"Laurino","sequence":"additional","affiliation":[{"name":"Institute of Clinical Physiology National Research Council  Pisa Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Francesco","family":"De Logu","sequence":"additional","affiliation":[{"name":"Department of Health Sciences Section of Clinical Pharmacology and Oncology University of Florence  Florence Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2026,9,23]]},"reference":[{"key":"e_1_2_9_2_1","doi-asserted-by":"crossref","first-page":"91","DOI":"10.1016\/0304-3959(94)00095-V","article-title":"The Formalin Test: Scoring Properties of the First and Second Phases of the Pain Response in Rats","volume":"60","author":"Abbott F. 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