{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,13]],"date-time":"2026-10-13T01:35:11Z","timestamp":1791855311299,"version":"4.3.4"},"reference-count":28,"publisher":"MDPI AG","issue":"6","license":[{"start":{"date-parts":[[2026,3,13]],"date-time":"2026-03-13T00:00:00Z","timestamp":1773360000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Diagnostics"],"abstract":"<jats:p>Background\/Objectives: This study aimed to quantitatively assess site-specific mean autofluorescence intensity across normal oral mucosal subsites and to evaluate the effectiveness of Autofluorescence Imaging (AFI) as an adjunct tool for distinguishing benign lesions, OPMDs, and oral cancers by comparing lesion intensity with anatomically matched healthy subsites. Methods: This observational study employed dual-mode imaging, comprising paired White Light Imaging (WLI) and AFI, captured from different oral cavity subsites using a smartphone-based point-of-care device. The Region of Interest (ROI) was annotated on WLI and automatically mapped to the corresponding AFI for both normal mucosa and lesions. WLI and AFI images were separated into their constituent red, green, and blue (RGB) channels, and AFI intensity was quantified via ImageJ. Results: A total of 1380 dual-mode images were acquired from 86 healthy participants. AFI intensities were comparable across most oral subsites, except for the lateral and ventral tongue. The lateral border showed the lowest fluorescence (Green channel-GC: 68.12 \u00b1 28.27; Blue channel-BC: 25.29 \u00b1 7.93), whereas the ventral tongue showed the highest (GC: 98.89 \u00b1 42.22; BC: 37.08 \u00b1 11.04; both p &lt; 0.001). Among 611 lesions, predominantly from the buccal mucosa, AFI intensity declined progressively with increasing disease severity. Homogeneous leukoplakia (n = 149; GC: 38.62 \u00b1 25.05; BC: 21.60 \u00b1 9.50), non-homogeneous leukoplakia (n = 25; GC: 30.42 \u00b1 18.66; BC: 18.25 \u00b1 7.17) and oral cancer (n = 21; GC: 23.39 \u00b1 15.53; BC: 15.82 \u00b1 7.15; all p &lt; 0.001) showed markedly reduced fluorescence, while benign lesions (n: 44; GC: 66.99 \u00b1 30.88; BC: 32.01 \u00b1 13.62) exhibited intermediate intensities, supporting AFI\u2019s discriminative potential. Conclusions: This phase-1, proof-of-concept study highlights subsite-specific variations in autofluorescence intensity within healthy oral mucosa, providing an essential baseline for objective interpretation of lesion-associated fluorescence changes. AFI has the potential to be used as a non-invasive adjunct for monitoring OPMDs. Further validation in larger and more diverse cohorts is required before clinical implementation.<\/jats:p>","DOI":"10.3390\/diagnostics16060857","type":"journal-article","created":{"date-parts":[[2026,3,13]],"date-time":"2026-03-13T11:12:25Z","timestamp":1773400345000},"page":"857","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Quantitative Autofluorescence Imaging of Oral Mucosa and Lesions: A Proof-of-Concept Study"],"prefix":"10.3390","volume":"16","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-4594-8674","authenticated-orcid":false,"given":"Keerthi","family":"Gurushanth","sequence":"first","affiliation":[{"name":"Department of Oral Medicine and Radiology, KLE Society\u2019s Institute of Dental Sciences, Bengaluru 560022, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sumsum P.","family":"Sunny","sequence":"additional","affiliation":[{"name":"Integrated Head and Neck Oncology Program, Mazumdar Shaw Medical Foundation, Narayana Health City, Bommsandra Industrial Area, Bengaluru 560099, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shubha","family":"Gurudath","sequence":"additional","affiliation":[{"name":"Department of Oral Medicine and Radiology, KLE Society\u2019s Institute of Dental Sciences, Bengaluru 560022, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Harshita","family":"Thakur","sequence":"additional","affiliation":[{"name":"Department of Oral Medicine and Radiology, KLE Society\u2019s Institute of Dental Sciences, Bengaluru 560022, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kripa","family":"Adlene Edith","sequence":"additional","affiliation":[{"name":"Department of Oral Medicine and Radiology, KLE Society\u2019s Institute of Dental Sciences, Bengaluru 560022, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Keerthi","family":"Krishnakumar","sequence":"additional","affiliation":[{"name":"Department of Oral Medicine and Radiology, KLE Society\u2019s Institute of Dental Sciences, Bengaluru 560022, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shikha","family":"Jha","sequence":"additional","affiliation":[{"name":"Department of Oral Medicine and Radiology, KLE Society\u2019s Institute of Dental Sciences, Bengaluru 560022, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5002-0355","authenticated-orcid":false,"given":"Pavitra","family":"Chandrashekhar","sequence":"additional","affiliation":[{"name":"Department of Oral Pathology, KLE Society\u2019s Institute of Dental Sciences, Bengaluru 560022, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Satyajit","family":"Topajiche","sequence":"additional","affiliation":[{"name":"Department of Oral Pathology, KLE Society\u2019s Institute of Dental Sciences, Bengaluru 560022, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lynette","family":"Linzbuoy","sequence":"additional","affiliation":[{"name":"Biocon Foundation, Bengaluru 560099, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sanjana","family":"Patrick","sequence":"additional","affiliation":[{"name":"Biocon Foundation, Bengaluru 560099, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0005-8550-7031","authenticated-orcid":false,"given":"Ramyashree","family":"Rao","sequence":"additional","affiliation":[{"name":"Department of Oral Medicine and Radiology, KLE Society\u2019s Institute of Dental Sciences, Bengaluru 560022, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Simranjeet","family":"Kaur","sequence":"additional","affiliation":[{"name":"Department of Oral Medicine and Radiology, KLE Society\u2019s Institute of Dental Sciences, Bengaluru 560022, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Umeshgouda","family":"Patil","sequence":"additional","affiliation":[{"name":"Department of Oral Medicine and Radiology, KLE Society\u2019s Institute of Dental Sciences, Bengaluru 560022, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ananya","family":"Nagaraj","sequence":"additional","affiliation":[{"name":"Department of Oral Medicine and Radiology, KLE Society\u2019s Institute of Dental Sciences, Bengaluru 560022, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bofan","family":"Song","sequence":"additional","affiliation":[{"name":"College of Optical Sciences, The University of Arizona, Tucson, AZ 85721, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rongguang","family":"Liang","sequence":"additional","affiliation":[{"name":"College of Optical Sciences, The University of Arizona, Tucson, AZ 85721, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shubhasini","family":"Raghavan","sequence":"additional","affiliation":[{"name":"Department of Oral Medicine and Radiology, KLE Society\u2019s Institute of Dental Sciences, Bengaluru 560022, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anupama","family":"Shetty","sequence":"additional","affiliation":[{"name":"Biocon Foundation, Bengaluru 560099, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Amritha","family":"Suresh","sequence":"additional","affiliation":[{"name":"Integrated Head and Neck Oncology Program, Mazumdar Shaw Medical Foundation, Narayana Health City, Bommsandra Industrial Area, Bengaluru 560099, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Moni Abraham","family":"Kuriakose","sequence":"additional","affiliation":[{"name":"Karkinos Healthcare, Kochi 682017, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5287-8323","authenticated-orcid":false,"given":"Praveen","family":"Birur Nagaraj","sequence":"additional","affiliation":[{"name":"Department of Oral Medicine and Radiology, KLE Society\u2019s Institute of Dental Sciences, Bengaluru 560022, India"},{"name":"Biocon Foundation, Bengaluru 560099, India"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,3,13]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"778","DOI":"10.1002\/ijc.33588","article-title":"Cancer statistics for the year 2020: An overview","volume":"149","author":"Ferlay","year":"2021","journal-title":"Int. J. Cancer"},{"key":"ref_2","first-page":"209","article-title":"Global Cancer Statistics 2020: GLOBOCAN Estimates of Incidence and Mortality Worldwide for 36 Cancers in 185 Countries","volume":"71","author":"Sung","year":"2021","journal-title":"CA Cancer J. Clin."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"107","DOI":"10.1146\/annurev-anchem-061020-123959","article-title":"Saliva Diagnostics","volume":"15","author":"Nonaka","year":"2022","journal-title":"Annu. Rev. Anal. Chem."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"14283","DOI":"10.1038\/s41598-022-18249-x","article-title":"Field validation of deep learning based Point-of-Care device for early detection of oral malignant and potentially malignant disorders","volume":"12","author":"Birur","year":"2022","journal-title":"Sci. Rep."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"53","DOI":"10.1016\/j.jdsr.2023.12.004","article-title":"Adjunctive aids for the detection of oral squamous cell carcinoma and oral potentially malignant disorders: A systematic review of systematic reviews","volume":"60","author":"Lau","year":"2024","journal-title":"Jpn. Dent. Sci. Rev."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"1490","DOI":"10.1111\/odi.15001","article-title":"Feasibility, Reliability, and Effectiveness of Oral cancer screening in South Asia and Southeast Asian countries: A systematic review and Meta-analysis","volume":"31","author":"Gurushanth","year":"2024","journal-title":"Oral Dis."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"539","DOI":"10.1002\/hed.26006","article-title":"Potentially malignant disorders of the oral cavity and oral dysplasia: A systematic review and meta-analysis of malignant transformation rate by subtype","volume":"42","author":"Iocca","year":"2020","journal-title":"Head Neck"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"20","DOI":"10.1016\/j.oraloncology.2017.03.003","article-title":"Novel quantitative analysis of autofluorescence images for oral cancer screening","volume":"68","author":"Huang","year":"2017","journal-title":"Oral Oncol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"101034","DOI":"10.1016\/j.disamonth.2020.101034","article-title":"Advanced diagnostic aids for oral cancer","volume":"66","author":"Madhura","year":"2020","journal-title":"Dis. Mon."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"105877","DOI":"10.1016\/j.oraloncology.2022.105877","article-title":"Non-invasive imaging of oral potentially malignant and malignant lesions: A systematic review and meta-analysis","volume":"130","author":"Mendonca","year":"2022","journal-title":"Oral Oncol."},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"Uthoff, R.D., Song, B., Sunny, S., Patrick, S., Suresh, A., Kolur, T., Keerthi, G., Spires, O., Anbarani, A., and Wilder-Smith, P. (2018). Point-of-care, smartphone-based, dual-modality, dual-view, oral cancer screening device with neural network classification for low-resource communities. PLoS ONE, 13.","DOI":"10.1371\/journal.pone.0207493"},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Uthoff, R.D., Wilder-Smith, P., Sunny, S., Suresh, A., Patrick, S., Anbarani, A., Liang, R., Song, B., Birur, P., and Kuriakose, M.A. (2018, January 27\u201328). Development of a Dual-Modality, Dual-View Smartphone-Based Imaging System for Oral Cancer Detection. Proceedings of the Design and Quality for Biomedical Technologies XI, San Francisco, CA, USA.","DOI":"10.1117\/12.2296435"},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Warnakulasuriya, S., and Greenspan, J.S. (2020). Textbook of Oral Cancer: Prevention, Diagnosis and Management, Springer.","DOI":"10.1007\/978-3-030-32316-5"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"3","DOI":"10.1016\/j.abd.2023.06.001","article-title":"Variations of oral anatomy and common oral lesions","volume":"99","author":"Souza","year":"2024","journal-title":"An. Bras. Dermatol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"816","DOI":"10.1007\/s12663-024-02198-1","article-title":"Holistic Approach for the Early Detection of Oral Cancer: A Comprehensive Training Module","volume":"23","author":"Gurushanth","year":"2024","journal-title":"J. Maxillofac. Oral Surg."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"717","DOI":"10.1111\/j.1601-0825.2010.01684.x","article-title":"Optical diagnostics in the oral cavity: An overview","volume":"16","author":"Holtzman","year":"2010","journal-title":"Oral Dis."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"29943","DOI":"10.1038\/srep29943","article-title":"Accuracy of autofluorescence in diagnosing oral squamous cell carcinoma and oral potentially malignant disorders: A comparative study with aero-digestive lesions","volume":"6","author":"Luo","year":"2016","journal-title":"Sci. Rep."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"491","DOI":"10.1111\/odi.13071","article-title":"Optical fluorescence imaging in oral cancer and potentially malignant disorders: A systematic review","volume":"26","author":"Tiwari","year":"2020","journal-title":"Oral Dis."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"562","DOI":"10.1002\/ar.23097","article-title":"Regional Quantitative Histological Variations in Human Oral Mucosa","volume":"298","author":"Ciano","year":"2015","journal-title":"Anat. Rec."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"239","DOI":"10.1364\/OE.3.000239","article-title":"In vivo OCT imaging of hard and soft tissue of the oral cavity","volume":"3","author":"Feldchtein","year":"1998","journal-title":"Opt. Express"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"460","DOI":"10.1097\/CEJ.0b013e32834fdb6d","article-title":"Clinical evaluation of an autofluorescence diagnostic device for oral cancer detection: A prospective randomized diagnostic study","volume":"21","author":"Rana","year":"2012","journal-title":"Eur. J. Cancer Prev."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"1012","DOI":"10.1016\/j.oraloncology.2014.07.002","article-title":"Assessment of a decision making protocol to improve the efficacy of VELscopeTM in general dental practice: A prospective evaluation","volume":"50","author":"Bhatia","year":"2014","journal-title":"Oral. Oncol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"856","DOI":"10.1002\/hed.21834","article-title":"Efficacy of tissue autofluorescence imaging (velscope) in the visualization of oral mucosal lesions","volume":"34","author":"Farah","year":"2012","journal-title":"Head Neck"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"2267","DOI":"10.1007\/s00784-015-1457-9","article-title":"Assessing the accuracy of autofluorescence, chemiluminescence and toluidine blue as diagnostic tools for oral potentially malignant disorders\u2014A clinicopathological evaluation","volume":"19","author":"Awan","year":"2015","journal-title":"Clin. Oral Investig."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"797","DOI":"10.1016\/j.adaj.2017.08.045","article-title":"Adjuncts for the evaluation of potentially malignant disorders in the oral cavity","volume":"148","author":"Lingen","year":"2017","journal-title":"J. Am. Dent. Assoc."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"423","DOI":"10.1158\/1940-6207.CAPR-08-0229","article-title":"Objective Detection and Delineation of Oral Neoplasia Using Autofluorescence Imaging","volume":"2","author":"Roblyer","year":"2009","journal-title":"Cancer Prev. Res."},{"key":"ref_27","first-page":"025001","article-title":"Computer-assisted discrimination of cancerous and pre-cancerous from benign oral lesions based on multispectral autofluorescence imaging endoscopy","volume":"1","author":"Sierra","year":"2024","journal-title":"Biophotonics Discov."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"024019","DOI":"10.1117\/1.2904658","article-title":"Multispectral optical imaging device for in vivo detection of oral neoplasia","volume":"13","author":"Roblyer","year":"2008","journal-title":"J. Biomed. 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