Evaluation of hβD-1 and hβD-2 levels in saliva of patients with oral mucosal diseases

Objective: This study aimed to determine a possible correlation between oral mucosal disease and salivary concentrations of the antimicrobial peptides human beta-defensin-1 (hβD-1) and human betadefensin- 2 (hβD-2). Method: The present work focussed on the establishment of a reversed phase-high performance liquid chromatography (RP-HPLC) procedure to quantify human beta-defensins (hβD-1 and hβD-2) in saliva samples of patients with oral diseases such as lichen planus (n = 10), Behçet (n = 10) and recurrent apthous stomatitis (n = 10). Results: Linear calibration range for hβD-1 and hβD-2 defensins was 1.67-200 μg mL-1 and 3.13 - 100 μg mL-1 with R2 values of 0.9998 and 0.996, correspondingly. The concentration of beta-defensins in saliva was determined by comparing the peak areas of eluted hβD-1 and hβD-2 with that of their standards. The variation of the amount of beta-defensins was evaluated by comparisons of the results obtained from the patients with oral mucosal diseases before and after treatments and the control subjects. The limit of detection (LOD) and limit of quantification (LOQ) were found to be 1.62 μg mL-1 and 5.39 μg mL-1 for hβD-1 and 0.94 μg mL-1 and 3.13 μg mL-1 for hβD-2, respectively. Conclusion: The salivary beta-defensin concentration was significantly higher in patients with oral mucosal diseases than in healthy volunteers; furthermore, in patients with oral mucosal diseases, the concentration was significantly higher before treatment than after treatment.