Biomarkers of Periodontitis in Clarified and Cellular Saliva

Objectives: To detect a potential correlation between the soluble and epithelial cell associated expression of TLR-2 and TLR-4 in saliva of chronic periodontitis patients.

Methods: Unstimulated whole saliva (UWS) will be collected from 20 patients with generalized chronic periodontitis and from 20 healthy individuals after obtaining an informed consent via IRB approved saliva bank 1209009585. Each saliva sample will be processed immediately to separate clarified and cellular saliva. Salivary epithelial cells (SECs) will be isolated from the cellular pellet of each UWS sample by centrifugation after lysing leukocytes with Zap-O-globin. The epithelial cell enriched pellet and the clarified saliva will be stored at -80°C until further analysis. Salivary sTLR-2 and sTLR-4 in clarified saliva will be measured by ELISA. The SECs will be assessed for TLR-2 and TLR-4 transcripts by quantitative real-time PCR.
Results: Anticipated result: salivary sTLR-2 and sTLR-4 will be inversely correlated with paired SECs associated TLR-2 or TLR-4 in chronic periodontitis. Significant differences will be observed in the soluble or epithelial cell associated TLR-2 and TLR-4 in saliva in individuals with chronic periodontitis and healthy individuals.
Conclusions: Collectively, the information in the literature suggests changes in soluble and membrane associated TLRs contribute to the ongoing pathogenesis of periodontitis. The proposed study aims at correlating the soluble and membrane associated TLR-2 and TLR-4 expressions in the same patient as indirect indicators of the current disease status in chronic periodontitis, in addition to the detection of established disease by conventional methods of diagnosing periodontitis.