Analysis of Salivary Proteins of Cleft Palate Patients

A cleft is a common birth deformity and a separation in a body's natural structure, usually formed before birth and occurs in the oral-facial region often involve the lip, the roof of the mouth (hard palate) or the soft tissue in the back of the mouth (soft palate).Children with a form of clefting are monitored by a cleft palate or craniofacial team through young adulthood. Complex treatment is needed and care can be lifelong.

Objectives: The aim of this study was the saliva proteome analysis of cleft palate patients for the reason of the better understanding and the early molecular diagnosis of this disease.

Methods: The salivary proteins were separated by using 1D-SDS-PAGE, and the tryptic peptides of the proteins were identified by matrix-assisted laser desorption ionization tandem time-of-flight (MALDI TOF/TOF) mass spectrometry. Reversed phase high performance liquid chromatography (RP-HPLC) coupled by electrospray ionization ion trap mass spectrometry (ESI IT MS/MS) was used for confirmation of the peptide mass fingerprint (PMF) results.

Results: Significant differences were detected on SDS gel between the control and the pathological samples. Saliva proteins were identified by using Mascot PMF and MS/MS database search engine. The presence of cleft palate stimulated the expression of several proteins, included novel keratinocyte-secreted proteins (e.g. dermokine), complement factor I and other potential biomarkers.

Conclusion: Analysis of body fluid proteome has become one of the most promising approaches to discovery of biomarkers for human diseases. Investigating the saliva proteome is especially advantageous due to the non-invasive sample collection. In this study MALDI TOF/TOF MS was used as a high-throughput analytical technique for identification of cleft palate stimulated proteins at the first time. In our further investigations we plan to analyze the identified cleft palate regulated proteins and their fragments in embryonic period for early molecular diagnosis of the disease.