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Objective: The aim of the sthudy is to sthudy the level of soluble Immune Checkpoint Molecules (B7.2, CTLA-4, Tim-3, Lag-3, PD-1) in the oral fluid during dental caries with the background of a lack and/or deficiency of 25-hydroxy-vitamin D in body., Materials and Methods: During the research 3 groups of people were formed, each one of them included 17 people aged from 20 to 24 years. The first group included students with high-intensity caries (above 9 DMFt index) and 25-hydroxy-vitamin D levels in blood serum >30 ng/ml, the second included students with high caries intensity and 25-hydroxy-vitamin D levels <30 ng/ml. The control group consisted of students with an average DMFt index of 1.5 (from 0 to 3) and a level of 25(OH)D in the blood more than 30 ng/ml. To determine the content of B7.2 (CD86), CTLA-4, Tim-3, Lag-3, PD-1, the Human Vascular Inflammation Panel 1 multiplex analysis kit from Biolegend (USA) was used., Results: The results of the research showed that during dental caries with a normal level of 25-hydroxy-vitamin D there are no significant changes in the content of Immune Checkpoint Molecules. With the background of deficiency and lack of 25-hydroxy-vitamin D there is a decrease in the amount of B7.2, LAG-3, Tim-3 and PD-1. These changes are being aggravated with an increase of the caries intensity., Conclusion: Vitamin D deficiency leads to a decrease in mucosal immunity of the oral cavity, the multiplication of pathogenic microorganisms, which in turn, releasing various metabolites, including cytokine-like substances, aggravate the pathological process and intensify carious lesions.

A simple and highly sensitive high-performance liquid chromatography assay is proposed to test salivary diamines (putrescine and cadaverine). Derivation was carried out with the orthophthalic aldehyde 2-mercaptoethanol. A rapid purification procedure for derivatives on the cartridges packed with 10 mg of hypercrosslinked polystyrene (Purosep-200) was first developed. Separation was made on a Chromolith (Merck), 100 x 4.6 mm in size, with monolithic reverse-phase silica gel (RP-18e) in the isocratic mode with ultraviolet (UV) detection at 230 nm. The eluent contained 55% acetonitrile and 45% 0.01 M phosphate buffer pH 6.8, added by 1% of isopropanol; flow rate was 1400 pl/min; pressure was 28 bars. Complete separation of diamine derivatives lasted at least 5 min. The sensitivity of the assay with UV detection (230 nm) was about 0.1 ngfor diamines and about 0.5 ng for the internal standard (IS) at a signal/noise ratio of 3.0, which enabled diamines to be determined in I pl (0.001 ml) of saliva. The simplicity, reproducibility, and high sensitivity of the assay along with the feasibility of its application on standard chromatographic equipment (an isocratic pump and an UV detector) make it suitable for routine clinical application.