Evaluation of salivary MMP-9 in oral squamous cell carcinoma and oral leukoplakia using ELISA

Abstract

Background: Cancer of the lip and the oral cavity is collectively the sixth most common malignancy worldwide, out of which 90% are oral squamous cell carcinomas (OSCCs). Oral cancer survival rates depend mainly upon the stage in which it is diagnosed. Successful early detection would eventually increase the survival rate. OSCCs may be preceded by potentially malignant disorders (PMDs) that are characterised by visible clinical changes in the oral mucosa. Correct diagnosis and timely treatment of PMDs may help prevent malignant transformation in oral lesions. Oral leukoplakia (OL) is the best known potentially malignant disorder of the oral mucosa with a malignant transformation rate of about 3% to 33%. Tumour markers in saliva have emerged as a new diagnostic tool in the early detection of oral cancer. Matrix metalloproteinase 9 (MMP-9) is a gelatinase which plays an important role in tumourogenisis. The present study was done to evaluate the salivary levels of MMP-9 in OSCC and oral leukoplakia patients using enzyme-linked immunosorbent assay (ELISA). Materials and Methods: The study was conducted among 102 subjects, which included 34 OSCC patients (group I), 34 OL patients (group II), and 34 healthy subjects (group III). Unstimulated saliva was collected by the passive drooling method from all the study subjects during the study period, centrifuged, and stored at -80°C. The salivary MMP-9 was estimated in mg/ml using the sandwich ELISA technique. The data were analysed using a statistical software package, EZR. One-way analysis of variance was used for the comparison of salivary MMP-9 levels in OSCC, OL, and normal oral mucosa. Scheffe’s multiple comparison was carried out to compare salivary MMP-9 levels among the different histological grades of OSCC and oral epithelial dysplasia. For all statistical interpretations, P ≤ 0.0 was considered the threshold for statistical significance. Results and Conclusion: The mean salivary MMP-9 level in OSCC, OL, and normal oral mucosa was 50.9 ± 5.7 ng/ml, 31.6 ± 6 ng/ml, and 16.2 ± 4.8 ng/ml, respectively. Patients with OSCC had significantly higher levels of salivary MMP-9 when compared to OL and normal mucosa. Higher levels of salivary MMP-9 were observed in poorly differentiated OSCC when compared to well and moderately differentiated OSCCs. The salivary MMP-9 was higher in severe oral epithelial dysplasia when compared to mild and moderate oral epithelial dysplasias. As malignant transformation rates are higher in patients with severe oral epithelial dysplasia when compared to mild and moderate oral epithelial dysplasia, salivary MMP-9 could be considered as a surrogate marker of malignant transformation.