{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,15]],"date-time":"2026-10-15T23:27:52Z","timestamp":1792106872757,"version":"4.4.0"},"reference-count":38,"publisher":"MDPI AG","issue":"1","license":[{"start":{"date-parts":[[2025,12,19]],"date-time":"2025-12-19T00:00:00Z","timestamp":1766102400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100004505","name":"University of Catania","doi-asserted-by":"crossref","id":[{"id":"10.13039\/501100004505","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Diagnostics"],"abstract":"<jats:p>Background\/Objectives: Immunohistochemistry (IHC) is a critical diagnostic tool in forensic pathology, enabling molecular-level assessment of wound vitality, post-mortem interval, and cause of death. However, IHC interpretation is subject to variability due to its reliance on human expertise. This study investigates whether artificial intelligence (AI), specifically a generative model, can assist in the diagnostic evaluation of IHC slides and replicate expert-level scoring, thereby improving consistency and reproducibility. Methods: A total of 225 high-resolution IHC images were classified into five immunoreactivity categories. The AI model (ChatGPT-4V) was trained on 150 labeled images and tested blindly on 75 unseen slides. Performance was assessed using confusion matrices, per-class precision\/recall\/F1, overall accuracy, Cohen\u2019s \u03ba (unweighted and weighted), and binary metrics (sensitivity, specificity, MCC). Results: Overall accuracy was 81.3% (95% CI: 71.1\u201388.5%), with substantial agreement (\u03ba = 0.767 unweighted; 0.805 linear-weighted; 0.848 quadratic-weighted). Binary classification achieved a sensitivity of 98.3%, specificity of 93.3%, MCC of 0.92. Accuracy was highest in extreme categories (\u2212 and +++, 93.3%), while intermediate classes (+ and ++) showed reduced performance (error rates up to 33%). Evaluation was rapid and consistent but lacked interpretative reasoning and struggled with borderline cases. Conclusions: AI-assisted diagnostic evaluation of IHC slides demonstrates promising accuracy and consistency, particularly in well-defined staining patterns. While not a replacement for human expertise, AI can serve as a valuable adjunct in forensic pathology, supporting rapid and standardized assessments. Ethical and legal considerations must guide its implementation in medico-legal contexts.<\/jats:p>","DOI":"10.3390\/diagnostics16010006","type":"journal-article","created":{"date-parts":[[2025,12,19]],"date-time":"2025-12-19T09:29:16Z","timestamp":1766136556000},"page":"6","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":8,"title":["AI-Assisted Diagnostic Evaluation of IHC in Forensic Pathology: A Comparative Study with Human Scoring"],"prefix":"10.3390","volume":"16","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-0357-8791","authenticated-orcid":false,"given":"Francesco","family":"Sessa","sequence":"first","affiliation":[{"name":"Department of Psychology and Health Sciences, Faculty of Human Sciences, Education, and Sports, Pegaso Telematic University, 80143 Naples, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0008-3027-1641","authenticated-orcid":false,"given":"Mara","family":"Ragusa","sequence":"additional","affiliation":[{"name":"Department of Medical, Surgical and Advanced Technologies \u201cG.F. Ingrassia\u201d, University of Catania, 95121 Catania, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7880-3450","authenticated-orcid":false,"given":"Massimiliano","family":"Esposito","sequence":"additional","affiliation":[{"name":"Faculty of Medicine and Surgery, \u201cKore\u201d University of Enna, 94100 Enna, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mario","family":"Chisari","sequence":"additional","affiliation":[{"name":"Faculty of Medicine and Surgery, \u201cKore\u201d University of Enna, 94100 Enna, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9895-3402","authenticated-orcid":false,"given":"Cristoforo","family":"Pomara","sequence":"additional","affiliation":[{"name":"Department of Medical, Surgical and Advanced Technologies \u201cG.F. Ingrassia\u201d, University of Catania, 95121 Catania, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Monica","family":"Salerno","sequence":"additional","affiliation":[{"name":"Department of Medical, Surgical and Advanced Technologies \u201cG.F. Ingrassia\u201d, University of Catania, 95121 Catania, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,12,19]]},"reference":[{"key":"ref_1","first-page":"666","article-title":"Application of Immunohistochemistry and Special Staining Technique in Forensic Traumatic Pathology Identification","volume":"37","author":"Chen","year":"2021","journal-title":"Fa Yi Xue Za Zhi"},{"key":"ref_2","doi-asserted-by":"crossref","unstructured":"Dervi\u0161evi\u0107, E., \u010camd\u017ei\u0107, N., Lazovi\u0107, E., Salihbegovi\u0107, A., Sessa, F., Spahovi\u0107, H., and D\u2019Errico, S. (2025). Histological and Immunohistochemical Evidence in Hypothermia-Related Death: An Experimental Study. Int. J. Mol. Sci., 26.","DOI":"10.3390\/ijms26157578"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1416","DOI":"10.1111\/iwj.13206","article-title":"Histopathological, Immunohistochemical, and Molecular Studies for Determination of Wound Age and Vitality in Rats","volume":"16","author":"Khalaf","year":"2019","journal-title":"Int. Wound J."},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"Salerno, M., Cocimano, G., Roccuzzo, S., Russo, I., Piombino-Mascali, D., M\u00e1rquez-Grant, N., Zammit, C., Esposito, M., and Sessa, F. (2022). New Trends in Immunohistochemical Methods to Estimate the Time since Death: A Review. Diagnostics, 12.","DOI":"10.3390\/diagnostics12092114"},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"Prangenberg, J., Doberentz, E., Mawick, A., and Madea, B. (2022). Mini Review: The Forensic Value of Heat Shock Proteins. Front. Med., 8.","DOI":"10.3389\/fmed.2021.800100"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"154","DOI":"10.1053\/j.semdp.2024.05.001","article-title":"Review of Immunohistochemistry Techniques: Applications, Current Status, and Future Perspectives","volume":"41","author":"Mebratie","year":"2024","journal-title":"Semin. Diagn. Pathol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"e335","DOI":"10.5858\/arpa.2023-0042-CP","article-title":"Recommendations for Performance Evaluation of Machine Learning in Pathology","volume":"148","author":"Hanna","year":"2024","journal-title":"Arch. Pathol. Lab. Med."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"185","DOI":"10.1097\/PAF.0000000000000408","article-title":"Usability of Immunohistochemistry in Forensic Samples with Varying Decomposition","volume":"39","author":"Lesnikova","year":"2018","journal-title":"Am. J. Forensic Med. Pathol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"27","DOI":"10.1186\/s41935-024-00401-5","article-title":"Is It Time for a Guideline on the Use of Immunohistochemistry in Forensic Pathology?","volume":"14","author":"Lancia","year":"2024","journal-title":"Egypt. J. Forensic Sci."},{"key":"ref_10","first-page":"354","article-title":"Immunohistochemistry as a Tool to Characterize Human Skin Wounds of Hanging Marks","volume":"26","author":"Legaz","year":"2018","journal-title":"Rom. J. Leg. Med."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"e26486","DOI":"10.1097\/MD.0000000000026486","article-title":"Immunohistochemistry in Postmortem Diagnosis of Acute Cerebral Hypoxia and Ischemia: A Systematic Review","volume":"100","author":"Barranco","year":"2021","journal-title":"Medicine"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"134","DOI":"10.1016\/S1344-6223(01)00020-7","article-title":"Immunohistochemistry of Pulmonary Surfactant-Associated Protein A in Acute Respiratory Distress Syndrome","volume":"3","author":"Zhu","year":"2001","journal-title":"Leg. Med."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"1564","DOI":"10.5858\/arpa.2014-0074-RA","article-title":"Standardization of Diagnostic Immunohistochemistry: Literature Review and Geisinger Experience","volume":"138","author":"Lin","year":"2014","journal-title":"Arch. Pathol. Lab. Med."},{"key":"ref_14","first-page":"5204","article-title":"Principle and Techniques of Immunohistochemistry\u2014A Review","volume":"6","author":"Kabiraj","year":"2015","journal-title":"Int. J. Biol. Med. Res."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"277","DOI":"10.1038\/s41698-024-00770-z","article-title":"A Universal Immunohistochemistry Analyzer for Generalizing AI-Driven Assessment of Immunohistochemistry across Immunostains and Cancer Types","volume":"8","author":"Brattoli","year":"2024","journal-title":"npj Precis. Oncol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"100408","DOI":"10.1016\/j.jpi.2024.100408","article-title":"Advancements in Pathology: Digital Transformation, Precision Medicine, and Beyond","volume":"16","author":"Ahuja","year":"2025","journal-title":"J. Pathol. Inform."},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Wong, A.N.N., He, Z., Leung, K.L., To, C.C.K., Wong, C.Y., Wong, S.C.C., Yoo, J.S., Chan, C.K.R., Chan, A.Z., and Lacambra, M.D. (2022). Current Developments of Artificial Intelligence in Digital Pathology and Its Future Clinical Applications in Gastrointestinal Cancers. Cancers, 14.","DOI":"10.3390\/cancers14153780"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"100688","DOI":"10.1016\/j.modpat.2024.100688","article-title":"Introduction to Artificial Intelligence and Machine Learning in Pathology and Medicine: Generative and Nongenerative Artificial Intelligence Basics","volume":"38","author":"Rashidi","year":"2025","journal-title":"Mod. Pathol."},{"key":"ref_19","first-page":"e70363","article-title":"Artificial Intelligence in Forensic Sciences: A Systematic Review of Past and Current Applications and Future Perspectives","volume":"16","author":"Ketsekioulafis","year":"2024","journal-title":"Cureus"},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Sessa, F., Esposito, M., Cocimano, G., Sablone, S., Karaboue, M.A.A., Chisari, M., Albano, D.G., and Salerno, M. (2024). Artificial Intelligence and Forensic Genetics: Current Applications and Future Perspectives. Appl. Sci., 14.","DOI":"10.3390\/app14052113"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"e190822207706","DOI":"10.2174\/2666484401666220819111603","article-title":"Artificial Intelligence: An Advanced Evolution in Forensic and Criminal Investigation","volume":"1","author":"Yadav","year":"2022","journal-title":"Curr. Forensic Sci."},{"key":"ref_22","first-page":"172","article-title":"Artificial Intelligence in Forensic Science","volume":"4","author":"Mohsin","year":"2023","journal-title":"Int. J. Forensic Res."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"1103","DOI":"10.1007\/s12024-023-00772-6","article-title":"The Integration and Implications of Artificial Intelligence in Forensic Science","volume":"20","author":"Tynan","year":"2024","journal-title":"Forensic Sci. Med. Pathol."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"844","DOI":"10.1038\/s41374-018-0057-0","article-title":"Principles and Approaches for Reproducible Scoring of Tissue Stains in Research","volume":"98","author":"Meyerholz","year":"2018","journal-title":"Lab. Investig."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"251","DOI":"10.1016\/j.diii.2024.04.003","article-title":"ChatGPT in Radiology: A Systematic Review of Performance, Pitfalls, and Future Perspectives","volume":"105","author":"Keshavarz","year":"2024","journal-title":"Diagn. Interv. Imaging"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"71","DOI":"10.1186\/s40164-024-00537-z","article-title":"Exploring Whether ChatGPT-4 with Image Analysis Capabilities Can Diagnose Osteosarcoma from X-Ray Images","volume":"13","author":"Ren","year":"2024","journal-title":"Exp. Hematol. Oncol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"1168","DOI":"10.1007\/s11604-024-01606-3","article-title":"Evaluating ChatGPT-4V in Chest CT Diagnostics: A Critical Image Interpretation Assessment","volume":"42","author":"Dehdab","year":"2024","journal-title":"Jpn. J. Radiol."},{"key":"ref_28","unstructured":"Cross, S., Furness, P., Igali, L., Snead, D., and Treanor, D. (2018). Best Practice Recommendations for Implementing Digital Pathology, The Royal College of Pathologists."},{"key":"ref_29","doi-asserted-by":"crossref","unstructured":"Wang, J., Zhang, S., Li, J., Deng, M., Zeng, Z., Dong, Z., Chen, F., Liu, W., Wu, L., and Yu, H. (2025). Development and Clinical Validation of Deep Learning-Based Immunohistochemistry Prediction Models for Subtyping and Staging of Gastrointestinal Cancers. BMC Gastroenterol., 25.","DOI":"10.1186\/s12876-025-04045-0"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"114227","DOI":"10.1016\/j.ejca.2024.114227","article-title":"Artificial Intelligence Breakthroughs in Pioneering Early Diagnosis and Precision Treatment of Breast Cancer: A Multimethod Study","volume":"209","author":"Darbandi","year":"2024","journal-title":"Eur. J. Cancer"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"109391","DOI":"10.1016\/j.compbiomed.2024.109391","article-title":"Artificial Intelligence for Breast Cancer Detection and Its Health Technology Assessment: A Scoping Review","volume":"184","author":"Uwimana","year":"2025","journal-title":"Comput. Biol. Med."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"1449","DOI":"10.1038\/s41379-022-01077-9","article-title":"Artificial Intelligence System Shows Performance at the Level of Uropathologists for the Detection and Grading of Prostate Cancer in Core Needle Biopsy: An Independent External Validation Study","volume":"35","author":"Jung","year":"2022","journal-title":"Mod. Pathol."},{"key":"ref_33","doi-asserted-by":"crossref","unstructured":"Sessa, F., Chisari, M., Esposito, M., Guardo, E., Di Mauro, L., Salerno, M., and Pomara, C. (2025). Advancing Diagnostic Tools in Forensic Science: The Role of Artificial Intelligence in Gunshot Wound Investigation\u2014A Systematic Review. Forensic Sci., 5.","DOI":"10.3390\/forensicsci5030030"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"445","DOI":"10.1007\/s00414-022-02928-5","article-title":"Applications of Artificial Intelligence in Forensic Sciences: Current Potential Benefits, Limitations and Perspectives","volume":"137","author":"Galante","year":"2022","journal-title":"Int. J. Leg. Med."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"52","DOI":"10.1016\/j.imed.2023.08.001","article-title":"Medical Artificial Intelligence and the Black Box Problem: A View Based on the Ethical Principle of \u201cDo No Harm\u201d","volume":"4","author":"Xu","year":"2024","journal-title":"Intell. Med."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"e354","DOI":"10.1002\/cpz1.354","article-title":"Ethical, Legal, and Social Implications (ELSI) Research: Methods and Approaches","volume":"2","author":"Ogbogu","year":"2022","journal-title":"Curr. Protoc."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"45","DOI":"10.1007\/s12559-023-10179-8","article-title":"Interpreting Black-Box Models: A Review on Explainable Artificial Intelligence","volume":"16","author":"Hassija","year":"2024","journal-title":"Cognit Comput."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"e10","DOI":"10.2196\/mhealth.4994","article-title":"Unpacking the Black Box: A Formative Research Approach to the Development of Theory-Driven, Evidence-Based, and Culturally Safe Text Messages in Mobile Health Interventions","volume":"4","author":"Maar","year":"2016","journal-title":"JMIR mHealth uHealth"}],"container-title":["Diagnostics"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2075-4418\/16\/1\/6\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,12,21]],"date-time":"2025-12-21T05:17:40Z","timestamp":1766294260000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2075-4418\/16\/1\/6"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,12,19]]},"references-count":38,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2026,1]]}},"alternative-id":["diagnostics16010006"],"URL":"https:\/\/doi.org\/10.3390\/diagnostics16010006","relation":{},"ISSN":["2075-4418"],"issn-type":[{"value":"2075-4418","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,12,19]]}}}