{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,10]],"date-time":"2026-10-10T14:54:59Z","timestamp":1791644099396,"version":"4.3.4"},"reference-count":54,"publisher":"MDPI AG","issue":"5","license":[{"start":{"date-parts":[[2026,2,27]],"date-time":"2026-02-27T00:00:00Z","timestamp":1772150400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Bladder cancer (BC) is the most common malignancy of the urinary tract. Approximately 75% of cases are non-muscle-invasive BC (NMIBC), while muscle-invasive BC (MIBC) and advanced tumors account for most cancer-specific mortality. Accurate assessment of tumor invasion is essential, as staging variability may lead to inappropriate treatments. Tumor invasion involves several mechanisms including extracellular matrix (ECM) remodeling mediated by metalloproteinases, angiogenesis, and cell adhesions. Masson\u2019s trichrome staining (MTS) provides relevant information on ECM composition. This study evaluated the application of machine learning to MTS-stained bladder biopsies to predict muscle invasion. A retrospective analysis of bladder biopsy images obtained from transurethral resections and cystectomies (2022\u20132024). A total of 702 histological images were analyzed. A convolutional neural network (CNN) was trained to classify tumors as MIBC or NMIBC and model outputs were correlated with clinical variables. The CNN achieved an accuracy of 95.2% in the training set and 90.1% in validation. Model-derived probabilities were significantly associated with tumor grade, lesion size, and muscle invasion. Logistic regression demonstrated a strong association with invasive disease (OR = 0.07, p = 0.017). CNN-based analysis of MTS-stained bladder biopsy images enable accurate prediction of muscle invasion, with potential to improve diagnostic precision.<\/jats:p>","DOI":"10.3390\/ijms27052237","type":"journal-article","created":{"date-parts":[[2026,2,27]],"date-time":"2026-02-27T10:41:02Z","timestamp":1772188862000},"page":"2237","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Use of Artificial Intelligence Model Associated with Masson\u2019s Trichrome Staining as a Predictor of Muscle Invasion in Bladder Cancer"],"prefix":"10.3390","volume":"27","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-0411-0672","authenticated-orcid":false,"given":"Diego","family":"Parrao","sequence":"first","affiliation":[{"name":"School of Medicine, Universidad de O\u2019Higgins, Rancagua 2820000, Chile"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1339-4358","authenticated-orcid":false,"given":"Hector","family":"Gallegos","sequence":"additional","affiliation":[{"name":"Department of Urology, Hospital S\u00f3tero del Rio, Santiago 8207257, Chile"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Karin","family":"Ruz","sequence":"additional","affiliation":[{"name":"Department of Pathology, Hospital Dr. Franco Ravera Zunino, Rancagua 2820000, Chile"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rom\u00e1n","family":"Lay","sequence":"additional","affiliation":[{"name":"School of Medicine, Universidad de O\u2019Higgins, Rancagua 2820000, Chile"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0002-9271-8070","authenticated-orcid":false,"given":"Catalina","family":"Saavedra","sequence":"additional","affiliation":[{"name":"School of Medicine, Universidad de O\u2019Higgins, Rancagua 2820000, Chile"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Renata","family":"Guerrero","sequence":"additional","affiliation":[{"name":"School of Medicine, Universidad de O\u2019Higgins, Rancagua 2820000, Chile"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0006-2146-5677","authenticated-orcid":false,"given":"Mat\u00edas","family":"Larra\u00f1aga","sequence":"additional","affiliation":[{"name":"Department of Urology, Hospital Dr. Franco Ravera Zunino, Rancagua 2820000, Chile"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Carolina B.","family":"Lindsay","sequence":"additional","affiliation":[{"name":"Research Department, Hospital Dr. Franco Ravera Zunino, Rancagua 2820000, Chile"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8581-7658","authenticated-orcid":false,"given":"Juan Crist\u00f3bal","family":"Bravo","sequence":"additional","affiliation":[{"name":"Department of Urology, Hospital Dr. Franco Ravera Zunino, Rancagua 2820000, Chile"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,2,27]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"58","DOI":"10.1038\/s41572-023-00468-9","article-title":"Bladder cancer","volume":"9","author":"Hansel","year":"2023","journal-title":"Nat. Rev. Dis. Primers"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"533","DOI":"10.1097\/JU.0000000000003846","article-title":"Diagnosis and Treatment of Non-Muscle Invasive Bladder Cancer: AUA\/SUO Guideline: 2024 Amendment","volume":"211","author":"Holzbeierlein","year":"2024","journal-title":"J. Urol."},{"key":"ref_3","first-page":"e27330","article-title":"A Review of the Etiology and Epidemiology of Bladder Cancer: All You Need to Know","volume":"14","author":"Halaseh","year":"2022","journal-title":"Cureus"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"309","DOI":"10.1016\/j.ajur.2024.08.005","article-title":"Bladder preservation alternatives in non-metastatic muscle-invasive bladder tumor: A systematic review and meta-analysis","volume":"12","author":"Parrao","year":"2025","journal-title":"Asian J. Urol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"2201","DOI":"10.3390\/curroncol31040163","article-title":"Active Surveillance in Non-Muscle Invasive Bladder Cancer, the Potential Role of Biomarkers: A Systematic Review","volume":"31","author":"Parrao","year":"2024","journal-title":"Curr. Oncol."},{"key":"ref_6","first-page":"372.e21","article-title":"Impact of pathologic re-review on grade, clinical stage, and risk stratification for patients with nonmuscle invasive bladder cancer","volume":"42","author":"Campbell","year":"2024","journal-title":"Urol. Oncol. Semin. Orig. Investig."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"1043","DOI":"10.1016\/j.euf.2024.06.007","article-title":"The Impact of Second Opinion Expert Pathology Review in Patient Management at the Time of Transurethral Resection of the Bladder","volume":"10","author":"Robesti","year":"2024","journal-title":"Eur. Urol. Focus."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"e307","DOI":"10.1016\/j.clgc.2016.01.002","article-title":"Role of Inter-Observer Variability and Quantification of Muscularis Propria in the Pathological Staging of Bladder Cancer","volume":"14","author":"Giunchi","year":"2016","journal-title":"Clin. Genitourin. Cancer"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"188","DOI":"10.5489\/cuaj.4126","article-title":"Pathology review impacts clinical management of patients with T1\u2013T2 bladder cancer","volume":"11","author":"Traboulsi","year":"2017","journal-title":"Can. Urol. Assoc. J."},{"key":"ref_10","first-page":"e143795","article-title":"Comparison of Interobserver Variability Between Two Grading Systems Used for Urothelial Carcinoma","volume":"16","author":"Jamshidi","year":"2024","journal-title":"Nephro-Urol. Mon."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1021","DOI":"10.1016\/j.juro.2016.06.049","article-title":"Diagnosis and Treatment of Non-Muscle Invasive Bladder Cancer: AUA\/SUO Guideline","volume":"196","author":"Chang","year":"2016","journal-title":"J. Urol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"270","DOI":"10.1177\/23523735241303350","article-title":"Repeat TURBT in large volume high-grade non-invasive bladder cancer","volume":"10","author":"Durant","year":"2024","journal-title":"Bladder Cancer"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"429","DOI":"10.4111\/kju.2015.56.6.429","article-title":"The clinical significance of a second transurethral resection for T1 high-grade bladder cancer: Results of a prospective study","volume":"56","author":"Shim","year":"2015","journal-title":"Korean J. Urol."},{"key":"ref_14","doi-asserted-by":"crossref","unstructured":"Ma, X., Zhang, Q., He, L., Liu, X., Xiao, Y., Hu, J., Cai, S., Cai, H., and Yu, B. (2024). Artificial intelligence application in the diagnosis and treatment of bladder cancer: Advance, challenges, and opportunities. Front. Oncol., 14.","DOI":"10.3389\/fonc.2024.1487676"},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Ferro, M., Falagario, U.G., Barone, B., Maggi, M., Crocetto, F., Busetto, G.M., del Giudice, F., Terracciano, D., Lucarelli, G., and Lasorsa, F. (2023). Artificial Intelligence in the Advanced Diagnosis of Bladder Cancer-Comprehensive Literature Review and Future Advancement. Diagnostics, 13.","DOI":"10.3390\/diagnostics13132308"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"925","DOI":"10.1016\/j.eururo.2018.02.014","article-title":"Repeat Transurethral Resection in Non\u2013muscle-invasive Bladder Cancer: A Systematic Review","volume":"73","author":"Cumberbatch","year":"2018","journal-title":"Eur. Urol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"1703","DOI":"10.1089\/end.2011.0081","article-title":"Repeated White Light Transurethral Resection of the Bladder in Nonmuscle-Invasive Urothelial Bladder Cancers: Systematic Review and Meta-Analysis","volume":"25","author":"Vianello","year":"2011","journal-title":"J. Endourol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"4047","DOI":"10.1007\/s00345-020-03569-2","article-title":"Follow-up in non-muscle invasive bladder cancer: Facts and future","volume":"39","author":"Witjes","year":"2021","journal-title":"World J. Urol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"BMI.S294","DOI":"10.4137\/BMI.S294","article-title":"The Role of Structural Extracellular Matrix Proteins in Urothelial Bladder Cancer (Review)","volume":"2","author":"Brunner","year":"2007","journal-title":"Biomark Insights"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"1999","DOI":"10.1002\/jum.16216","article-title":"Contrast-enhanced Ultrasound Combined With Elastography for the Evaluation of Muscle-invasive Bladder Cancer in Rats","volume":"42","author":"Hu","year":"2023","journal-title":"J. Ultrasound Med."},{"key":"ref_21","doi-asserted-by":"crossref","unstructured":"Dozmorov, M.G., Kyker, K.D., Saban, R., Knowlton, N., Dozmorov, I., Centola, M.B., and Hurst, R.E. (2006). Analysis of the interaction of extracellular matrix and phenotype of bladder cancer cells. BMC Cancer, 6.","DOI":"10.1186\/1471-2407-6-12"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"36128","DOI":"10.1038\/srep36128","article-title":"Linearized texture of three-dimensional extracellular matrix is mandatory for bladder cancer cell invasion","volume":"6","author":"Alfano","year":"2016","journal-title":"Sci. Rep."},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Bauman, T.M., Nicholson, T.M., Abler, L.L., Eliceiri, K.W., Huang, W., Vezina, C.M., and Ricke, W.A. (2014). Characterization of Fibrillar Collagens and Extracellular Matrix of Glandular Benign Prostatic Hyperplasia Nodules. PLoS ONE, 9.","DOI":"10.1371\/journal.pone.0109102"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"2393","DOI":"10.1007\/s00403-023-02628-z","article-title":"Differential expression of extracellular matrix proteins in the lesional skin of vitiligo patients","volume":"315","author":"Rani","year":"2023","journal-title":"Arch. Dermatol. Res."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"26731","DOI":"10.1007\/s11042-022-14305-w","article-title":"Machine learning and deep learning approach for medical image analysis: Diagnosis to detection","volume":"82","author":"Rana","year":"2023","journal-title":"Multimed. Tools Appl."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"256","DOI":"10.1016\/j.eururo.2020.04.023","article-title":"Deep Learning Predicts Molecular Subtype of Muscle-invasive Bladder Cancer from Conventional Histopathological Slides","volume":"78","author":"Woerl","year":"2020","journal-title":"Eur. Urol."},{"key":"ref_27","unstructured":"(2026). Applications of AI in urology. Nat. Rev. Urol., 23, 1."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"223","DOI":"10.1186\/s12943-024-02140-6","article-title":"Artificial intelligence alphafold model for molecular biology and drug discovery: A machine-learning-driven informatics investigation","volume":"23","author":"Guo","year":"2024","journal-title":"Mol. Cancer"},{"key":"ref_29","doi-asserted-by":"crossref","unstructured":"Guo, S.-B., Liu, D.-Y., Fang, X.-J., Meng, Y., Zhou, Z.-Z., Li, J., Li, M., Luo, L.-L., Li, H.-L., and Cai, X.-Y. (2025). Current concerns and future directions of large language model chatGPT in medicine: A machine-learning-driven global-scale bibliometric analysis. Int. J. Surg., ahead of print.","DOI":"10.1097\/JS9.0000000000003668"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"317","DOI":"10.1016\/j.ebiom.2017.12.026","article-title":"Deep Convolutional Neural Networks Enable Discrimination of Heterogeneous Digital Pathology Images","volume":"27","author":"Khosravi","year":"2018","journal-title":"EBioMedicine"},{"key":"ref_31","doi-asserted-by":"crossref","unstructured":"Yin, P.-N., Kc, K., Wei, S., Yu, Q., Li, R., Haake, A.R., Miyamoto, H., and Cui, F. (2020). Histopathological distinction of non-invasive and invasive bladder cancers using machine learning approaches. BMC Med. Inform. Decis. Mak., 20.","DOI":"10.1186\/s12911-020-01185-z"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"15330338241257479","DOI":"10.1177\/15330338241257479","article-title":"Detection of Muscularis propria Invasion in Urothelial Carcinoma Using Artificial Intelligence","volume":"23","author":"Fahoum","year":"2024","journal-title":"Technol. Cancer Res. Treat."},{"key":"ref_33","doi-asserted-by":"crossref","unstructured":"Islam, M.A., and Kumar, S. (2026). Masson\u2019s Trichrome Staining Technique to Evaluate Tissue Fibrosis, Springer.","DOI":"10.1007\/978-1-0716-4901-5_9"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"767","DOI":"10.1016\/S0929-6646(08)60189-8","article-title":"Masson Trichrome Stain Helps Differentiate Myofibroma from Smooth Muscle Lesions in the Head and Neck Region","volume":"107","author":"Chang","year":"2008","journal-title":"J. Formos. Med. Assoc."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"82609","DOI":"10.18632\/oncotarget.12089","article-title":"Positive association of collagen type I with non-muscle invasive bladder cancer progression","volume":"7","author":"Brooks","year":"2016","journal-title":"Oncotarget"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"3441","DOI":"10.2147\/OTT.S194568","article-title":"Collagen stiffness promoted non-muscle-invasive bladder cancer progression to muscle-invasive bladder cancer","volume":"12","author":"Zhu","year":"2019","journal-title":"OncoTargets Ther."},{"key":"ref_37","first-page":"100041","article-title":"Artificial-Intelligence-Driven Precision Medicine in Cancer Treatment","volume":"1","author":"Hassane","year":"2024","journal-title":"Prem. J. Sci."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"e12048","DOI":"10.1002\/inmd.12048","article-title":"Advances in AI-based Cancer Cytopathology (3\/2023)","volume":"1","author":"Yang","year":"2023","journal-title":"Interdiscip. Med."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"105209","DOI":"10.1016\/j.compbiomed.2022.105209","article-title":"AI-based Carcinoma Detection and Classification Using Histopathological Images: A Systematic Review","volume":"142","author":"Prabhua","year":"2022","journal-title":"Comput. Biol. Med."},{"key":"ref_40","doi-asserted-by":"crossref","unstructured":"Moghaddam, F.S., Dwabe, S., Mar, N., Safdari, L., Sabharwal, N., Goldberg, H., Daneshvar, M., and Kalebasty, A.R. (2024). The Role of Maximal TURBT in Muscle-Invasive Bladder Cancer: Balancing Benefits in Bladder Preservation and Beyond. Cancers, 16.","DOI":"10.3390\/cancers16193361"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"131","DOI":"10.1016\/j.urology.2018.10.027","article-title":"The Perioperative Morbidity of Transurethral Resection of Bladder Tumor: Implications for Quality Improvement","volume":"125","author":"Pereira","year":"2019","journal-title":"Urology"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"257","DOI":"10.1097\/CU9.0000000000000160","article-title":"A retrospective analysis of the factors associated with increased risk of readmission within 30 days after primary transurethral resection of bladder tumor","volume":"17","author":"Jindal","year":"2023","journal-title":"Curr. Urol."},{"key":"ref_43","doi-asserted-by":"crossref","unstructured":"Yang, X., Zhang, S., Cui, Y., Li, Y., Song, X., and Pang, J. (2023). Efficacy and safety of transurethral resection of bladder tumour combined with chemotherapy and immunotherapy in bladder-sparing therapy in patients with T1 high-grade or T2 bladder cancer: A protocol for a randomized controlled trial. BMC Cancer, 23.","DOI":"10.1186\/s12885-023-10798-2"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"1039","DOI":"10.1016\/j.eururo.2012.02.028","article-title":"Radical Cystectomy for Urothelial Carcinoma of the Bladder Without Neoadjuvant or Adjuvant Therapy: Long-Term Results in 1100 Patients","volume":"61","author":"Hautmann","year":"2012","journal-title":"Eur. Urol."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"e043266","DOI":"10.1136\/bmjopen-2020-043266","article-title":"Short-term morbidity and mortality following radical cystectomy: A systematic review","volume":"11","author":"Maibom","year":"2021","journal-title":"BMJ Open"},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"249","DOI":"10.1016\/j.ucl.2017.12.008","article-title":"Quality of Life After Radical Cystectomy","volume":"45","author":"Tyson","year":"2018","journal-title":"Urol. Clin. N. Am."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"281","DOI":"10.1016\/j.suronc.2016.05.027","article-title":"A systematic review and meta-analysis of quality of life outcomes after radical cystectomy for bladder cancer","volume":"25","author":"Yang","year":"2016","journal-title":"Surg. Oncol."},{"key":"ref_48","doi-asserted-by":"crossref","unstructured":"Bizzarri, F.P., Campetella, M., Recupero, S.M., Bellavia, F., D\u2019amico, L., Rossi, F., Gavi, F., Filomena, G.B., Russo, P., and Palermo, G. (2025). Female Sexual Function After Radical Treatment for MIBC: A Systematic Review. J. Pers. Med., 15.","DOI":"10.3390\/jpm15090415"},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"837","DOI":"10.1007\/s41669-024-00512-8","article-title":"Economic Impact of Bladder Cancer in the USA","volume":"8","author":"Clark","year":"2024","journal-title":"Pharmacoecon. Open"},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"536","DOI":"10.1016\/j.eururo.2024.12.002","article-title":"The Financial Burden of Localized and Metastatic Bladder Cancer","volume":"87","author":"Scilipoti","year":"2025","journal-title":"Eur. Urol."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"54","DOI":"10.1111\/bju.13265","article-title":"Repeat transurethral resection for non-muscle-invasive bladder cancer: A contemporary series","volume":"117","author":"Gendy","year":"2016","journal-title":"BJU Int."},{"key":"ref_52","doi-asserted-by":"crossref","unstructured":"Contieri, R., Lughezzani, G., Buffi, N.M., Taverna, G., Giacobbe, A., Micheli, E., Barra, S., Colombo, P., Vanni, E., and Guazzoni, G. (2022). Could We Safely Avoid a Second Resection in Selected Patients with T1 Non-Muscle-Invasive Bladder Cancer? Preliminary Results of Cost-Effectiveness Study from HUmanitas New Indications for ReTUR (HuNIRe) Multicenter Prospective Trial. Front. Oncol., 12.","DOI":"10.3389\/fonc.2022.879399"},{"key":"ref_53","first-page":"469.e1","article-title":"National complication and cost burden of transurethral resection of bladder tumor for bladder cancer","volume":"43","author":"Zhang","year":"2025","journal-title":"Urol. Oncol. Semin. Orig. Investig."},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"359","DOI":"10.1007\/s00345-025-05714-1","article-title":"Outpatient transurethral resection of bladder tumors: A systematic review of oncologic and safety outcomes","volume":"43","author":"Pattou","year":"2025","journal-title":"World J. Urol."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/27\/5\/2237\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,2,27]],"date-time":"2026-02-27T10:42:32Z","timestamp":1772188952000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/27\/5\/2237"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,2,27]]},"references-count":54,"journal-issue":{"issue":"5","published-online":{"date-parts":[[2026,3]]}},"alternative-id":["ijms27052237"],"URL":"https:\/\/doi.org\/10.3390\/ijms27052237","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,2,27]]}}}