Abstract
Oral Epithelial Dysplasia (OED) can be a precancerous alteration of the oral epithelium that may be a precursor to oral squamous cell carcinoma (OSCC). This study assessed the diagnostic accuracy of a salivary biomarker panel for early detection and risk stratification of OED. A cross-sectional study (september 2022 to February 2023) included 120 participants using non-probability consecutive sampling method. Unstimulated saliva samples were collected under standard conditions and tested for a set of markers, including, IL-6, IL-8, MMP-9, LDH, and Cyfra 21-1, by enzyme-linked immunosorbent assay (ELISA). The reference standard was histopathological assessment of the lesions for diagnosis of benign, mild dysplasia, moderate/severe dysplasia and oral squamous cell carcinoma. The data was analysed statistically using SPSS version 26.0. A chi-square testing and logistic regression were used, p≤0.05 was considered statistically significant. Mean age was 47.9 ± 11.4 years and 61.7% of the study population were male. Histopathological evaluation showed that the diagnosis of OED was confirmed in 39.2% while the diagnosis of oral squamous cell carcinoma was confirmed in 18.3% of the cases. Salivary levels of IL-6, IL-8, MMP-9, LDH and Cyfra 21-1 significantly rose with the severity of the histopathology (p<0.001). An association between high-risk salivary biomarker profiles and moderate to severe dysplasia and malignant transformation was found (χ²=29.84, p<0.001). The combined panel yielded sensitivity 89.2% (95% CI: 82.4–94.1), specificity 84.6% (95% CI: 76.3–91.0), and AUC 0.91 (95% CI: 0.86–0.95). The combined salivary biomarker panel demonstrated high diagnostic accuracy for identifying OED and distinguishing high-risk lesions from benign oral mucosal conditions.
References
1.Odell E, Kujan O, Warnakulasuriya S, Sloan P. Oral epithelial dysplasia: recognition, grading and clinical significance. Oral Diseases. 2021;27(8):1947-76.
2.Woo SB. Oral epithelial dysplasia and premalignancy. Head and neck pathology. 2019;13(3):423-39.
3.Tilakaratne WM, Jayasooriya PR, Jayasuriya NS, De Silva RK. Oral epithelial dysplasia: Causes, quantification, prognosis, and management challenges. Periodontology. 2019;80(1):126-47.
4.Liew YX, Kadir K, Phan CW, Leong KH, Kallarakkal TG, Tilakaratne WM. Salivary biomarkers: Effective diagnostic tool for oral leukoplakia and oral squamous cell carcinoma. Oral Diseases. 2024;30(6):3591-9.
5.Melguizo-Rodríguez L, Costela-Ruiz VJ, Manzano-Moreno FJ, Ruiz C, Illescas-Montes R. Salivary biomarkers and their application in the diagnosis and monitoring of the most common oral pathologies. International journal of molecular sciences. 2020;21(14):5173.
6.Babiuch K, Kuśnierz-Cabala B, Kęsek B, Okoń K, Darczuk D, Chomyszyn-Gajewska M. Evaluation of proinflammatory, NF-kappaB dependent cytokines: IL-1α, IL-6, IL-8, and TNF-α in tissue specimens and saliva of patients with oral squamous cell carcinoma and oral potentially malignant disorders. Journal of clinical medicine. 2020;9(3):867.
7.Pazhani J, Chanthu K, Jayaraman S, Varun BR. Evaluation of salivary MMP-9 in oral squamous cell carcinoma and oral leukoplakia using ELISA. Journal of Oral and Maxillofacial Pathology. 2023;27(4):649-54.
8.Javaraiah RK, David CM, Namitha J, Tiwari R, Benakanal P. Evaluation of salivary lactate dehydrogenase as a prognostic biomarker in tobacco users with and without potentially malignant disorders of the oral cavity. South Asian Journal of Cancer. 2020;9(02):093-8.
9.Rathore AS, Katyal S, Jain A, Shetty DC. Biochemical analysis of cytokeratin fragment 21-1 concentration and expression of cytokeratin 19 in oral potentially malignant disorders. Journal of Cancer Research and Therapeutics. 2020;16(3):452-7.
10.Jafari M, Hasanzadeh M. Non-invasive bioassay of Cytokeratin Fragment 21.1 (Cyfra 21.1) protein in human saliva samples using immunoreaction method: An efficient platform for early-stage diagnosis of oral cancer based on biomedicine. Biomedicine & Pharmacotherapy. 2020;131:110671.
11.Motahari P, Pournaghi-Azar F, Katebi K, Salehnia F, Abedi N. Evaluation of salivary biomarkers in patients with oral epithelial dysplasia: A systematic review. Journal of Oral Health and Oral Epidemiology. 2021;10(4):175-83.
12.Kaur J, Preethi M, Srivastava R, Borse V. Role of IL-6 and IL-8 biomarkers for optical and electrochemical based point-of-care detection of oral cancer. Biosensors and Bioelectronics: X. 2022;11:100212.
13.Piyarathne NS, Weerasekera MM, Fonseka PF, Karunatilleke AH, Liyanage RL, Jayasinghe RD, De Silva K, Yasawardene S, Gupta E, Jayasinghe JA, Abu-Eid R. Salivary interleukin levels in oral squamous cell carcinoma and oral epithelial dysplasia: findings from a Sri Lankan study. Cancers. 2023;15(5):1510.
14.Smriti K, Ray M, Chatterjee T, Shenoy RP, Gadicherla S, Pentapati KC, Rustaqi N. Salivary MMP-9 as a biomarker for the diagnosis of oral potentially malignant disorders and oral squamous cell carcinoma. Asian Pacific journal of cancer prevention: APJCP. 2020;21(1):233.
15.Deshpande SM, Choudhary S, Patankar S, Khetan P, Sadaphule G, Sachdev SS. Comparative analysis of Ki-67, α-SMA, and MMP-9 expression in oral submucous fibrosis and oral leukoplakia with/without dysplasia: insights into malignant transformation mechanisms. Revista Española De Patología. 2025;58(2):100804.
16.Miguel AF, Embaló B, Dias HB, Rivero ER. Immunohistochemical expression of MMP-9, TIMP-1, and vimentin and its correlation with inflammatory reaction and clinical parameters in oral epithelial dysplasia. Applied Immunohistochemistry & Molecular Morphology. 2021;29(5):382-9.
17.Iglesias‐Velázquez Ó, López‐Pintor RM, González‐Serrano J, Casanas E, Torres J, Hernandez G. Salivary LDH in oral cancer and potentially malignant disorders: A systematic review and meta‐analysis. Oral diseases. 2022;28(1):44-56.
18.Anthwal N, Gupta S, Begum S. Estimation of serum lactate dehydrogenase in oral leukoplakia and oral submucous fibrosis. Journal of Advanced Medical and Dental Sciences Research. 2020;8(10):162-4.
19.López-Pintor RM, González-Serrano J, Vallina C, Ivaylova Serkedzhieva K, Virto L, Nuevo P, Caponio VC, Iniesta M, Rodríguez Santamarta T, Lequerica Fernández P, Iglesias Velázquez Ó. Factors influencing salivary lactate dehydrogenase levels in oral squamous cell carcinoma and oral potentially malignant disorders. Frontiers in Oral Health. 2025;5:1525936.
20.Liang Y, Yi Z, Li J, Ye J. The diagnostic value of CYFRA 21–1 in oral squamous cell carcinoma: a meta-analysis. Expert Review of Anticancer Therapy. 2024;24(11):1161-8.
21.Adusumilli P, Babburi S, Venigalla A, Benarji KA, Sai SK, Soujanya P. Estimation of salivary and serum CYFRA 21-1 levels in patients with oral squamous cell carcinoma. Journal of Oral and Maxillofacial Pathology. 2023;27(1):98-102.
22.AlAli AM, Walsh T, Maranzano M. CYFRA 21-1 and MMP-9 as salivary biomarkers for the detection of oral squamous cell carcinoma: a systematic review of diagnostic test accuracy. International journal of oral and maxillofacial surgery. 2020;49(8):973-83.
23.Sayed AB, Hendre A, Gudur A, Kakade S. Role of Cytokeratin 19 Fragment Antigen 21-1 (CYFRA 21-1) in Early Diagnosis of Oral Squamous Cell Carcinoma. Journal of Datta Meghe Institute of Medical Sciences University. 2024;19(3):469-72.
24.Turton N, Payne K, Higginson J, Praveen P, Mehanna H, Nankivell P. Prognostic biomarkers for malignant progression of oral epithelial dysplasia: an updated systematic review and meta-analysis. British Journal of Oral and Maxillofacial Surgery. 2024;62(5):415-25.
25.de Morais EF, Pinheiro JC, Lira JA, Mafra RP, Barboza CA, de Souza LB, de Almeida Freitas R. Prognostic value of the immunohistochemical detection of epithelial-mesenchymal transition biomarkers in oral epithelial dysplasia: A systematic review. Medicina Oral, Patología Oral y Cirugía Bucal. 2020;25(2):e205.
26.Peng J, Xu Z, Dan H, Li J, Wang J, Luo X, Xu H, Zeng X, Chen Q. Oral epithelial dysplasia detection and grading in oral leukoplakia using deep learning. BMC Oral Health. 2024;24(1):434.
27.Ghalwash D, Ammar A, Abou-Bakr A, Diab AH, El-Gawish A. Validation of salivary proteomic biomarkers for early detection of oral cancer in the Egyptian population. Future Science OA. 2025;11(1):2432222.
28.Huang Y, Zhang Q, Guo Z, Deng G, Chen R, Zheng Y. Potential noninvasive biomarkers for the malignant transformation of oral leukoplakia: A systematic review and meta‐analysis. Cancer Medicine. 2023;12(13):14718-30.
29.Sivadasan P, Gupta MK, Sathe G, Sudheendra HV, Sunny SP, Renu D, Hari PS, Gowda H, Suresh A, Kuriakose MA, Sirdeshmukh R. Salivary proteins from dysplastic leukoplakia and oral squamous cell carcinoma and their potential for early detection. Journal of Proteomics. 2020;212:103574.
Piyarathne NS, Rasnayake RM, Angammana R, Chandrasekera P, Ramachandra S, Weerasekera M, Yasawardene S, Abu-Eid R, Jayasinghe JP, Gupta E. Diagnostic salivary biomarkers in oral cancer and oral potentially malignant disorders and their relationships to risk factors–A systematic review. Expert Review of Molecular Diagnostics. 2021;21(8):789-807.
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