Analysis of changes in biochemical parameters of oral liquid under the influence of lecithin-calcium complex in young patients with multiple caries

Given the high prevalence of dental caries, it is important to find new means and methods of prevention. At present preparations of natural origin, such as lecithin and calcium citrate, are widely used. The aim was to study the lecithin-calcium complex effect on the oral fluid biochemical parameters of individuals with multiple dental caries. Materials and methods. 85 KhNMU Dental Faculty students-volunteers were examined. Groups of participants were formed depending on the dental caries intensity level index value for the oral fluid biochemical parameters determination (degree of oral dysbiosis, antioxidant-prooxidant index) in patients before and after the start of the lecithin-calcium complex treatment course. Results. In persons with multiple dental caries the degree of dysbiosis is 3 times higher than normal, the concentration of calcium decreases by 1.5 times and the antioxidant-prooxidant index reduces by 2 times. After lecithin-calcium complex treatment course the dysbiosis degree decreases almost by 8 times, the concentration of calcium and API return to normal. This indicates the ability of the lecithin-calcium complex to regulate the oral cavity microbiocenosis and restore antioxidant-prooxidant indices. Conclusions. In patients with multiple dental caries the phenomena of the oral cavity dysbiosis, a decrease in the protective antioxidant system level and a decrease in the oral fluid calcium concentration are observed. The lecithin-calcium complex intake during the month completely eliminated the phenomenon of dysbiosis in the oral cavity, increased the antioxidant-pro oxidant system level and the oral fluid calcium concentration to normal.

The purpose of this work was to study the effect of calcium citrate containing complex and as a source of phosphoruslecithin on the oral fluid biochemical indicators in persons with multiple dental caries.
The choice of biochemical parameters of the oral fluid was due to the fact that the condition of latter largely determines the carious lesion pathogenesis [1][2][3][4][5]. Among the selected biochemical parameters were inflammation markers (MDA, TPA), a member of the remineralization system -calcium, one of the antioxidant system factorsenzyme catalase, and finally, enzymatic indicators of oral microbiocenosis state -urease and lysozyme.

Materials and methods
85 Kharkiv National Medical University 2, 3 and 5 courses Dental Faculty students-volunteers were examined. A survey of volunteer students in the clinic was conducted under standard conditions of the dental office by questioning and objective clinical oral cavity assessment with the use of diagnostic dental instruments. The condition of the oral cavity was examined according to the method proposed by WHO. When collecting anamnesis, attention was focused on harmful habits related to dental health, including the frequency of carbohydrate-containing foods consumption, irregular oral hygiene and low level of motivation for dental pathology. Patients underwent initial clinical examination and oral cavity sanation with subsequent dynamic observation (after 6 and 12 months).
The analysis of the clinical examination results made it possible to form three groups of participants depending on the level of dental caries intensity (LCI) index value, which was performed using the index of carious tooth decay intensity -CFE index (C means the number of carious teeth, F -the number of sealed teeth, E -the number of removed or to be removed teeth), with division by patient's age and interpretation according to Leus P. A. recommendations. For the adult population LCI <0.15 -low and LCI from 0.15 to 0.30 -average; LCI from 0.31 to 0.60 -high and LCI >0.60 indicates a very high level of caries intensity.
The control group consisted of 12 apparently healthy patients, without severe dental and somatic pathology, with LCI from 0 to 0.30. Criteria for patients with caries selection, based on objective clinical examination data, were the presence of a high and very high level of tooth decay (0.31 to 0.60 and above 0.60). Basic group included 52 patients in initial state and 21 patients in one month of lecithin-calcium complex treatment.
Patients of the study groups received a scheme of complex hygienic and preventive measures, which included the observance of rational and balanced diet, the rules of oral hygiene and hygiene control every two months during the year of observation. For individual oral hygiene throughout the year, Colgate toothpaste, "Triple Action with Fluoride" with an active fluoride concentration of 1450 ppm F (1.1 %) twice a day, a toothbrush with a high cleaning index, dental floss, "Colgate Plax" softener complex action "Tender mint" containing sodium fluoride (0.025 %), which was applied twice a day after tooth brushing within a month twice a year.
Additionally, from the first day of the examination patients of the basic group received a tableted lecithin-calcium complex "Lecithin-2", manufactured by the NPA "Odesa Biotechnology" (TU U 15.8-13903778-82-2000) [3]. The composition of the drug includes: sunflower lecithin and calcium citrate in a ratio of 1:1. Patients took daily dose of 600 mg of lecithin-calcium complex three times a day for half an hour before eating 1 tablet until complete resorption  in the oral cavity. The course of prevention was carried out within one month twice a year. In patients on the first day of the examination, and then a month later, unstimulated saliva (morning on an empty stomach) was collected in accordance with the recommendations [6].
To compare the indices, the oral liquid was as practically healthy, without pronounced dental and somatic pathology.
The relative activities of urease and lysozyme ratio was used to calculate the degree of the oral cavity dysbiosis according to the enzymatic method of A. Levitsky [15], and the antioxidant-prooxidant index (API) was calculated as the ratio of catalase activity and MDA concentration [7].
Statistical processing of data was carried out using the licensed package of programs Biostatistics v.4.03 and Statistica v.5.0, the obtained results reliability was assessed by the Student's t-test, with a critical significance level P ≤ 0.05. Table 1 presents the results of oral fluid biochemical parameters determination in patients before and after 1 month of the lecithin-calcium complex intake, as well as in healthy people. As it can be seen from these data, only two markers of inflammation (MDA and TPA) increase significantly in persons with caries and also reliably return to normal after the lecithin-calcium complex treatment.

Results and discussion
In patients with caries, the oral fluid calcium concentration is significantly 1.5 times reduced, but returns to normal after the course of treatment, which indicates the restoration of saliva remineralizing function.
In patients with tooth decay the catalase activity as one of antioxidant enzymes decreases by more than 2 times, and in a month of the lecithin-calcium complex intake this figure almost returns to normal. Table 2 presents the results of urease (reflecting the degree of microbial contamination) and lysozyme (the most important factor of nonspecific immunity) activity determination, and also their relative activities are calculated, which are necessary for the oral cavity dysbiosis degree determination.
As it can be seen from these data, the activity of urease in the oral fluid of patients is slightly higher than normal (however, P > 0.05), and after the lecithin-calcium complex treatment the activity of urease decreases by 5 times, which indicates a decrease in the oral cavity microbial contamination. On the contrary, the activity of lysozyme in the oral fluid of patients with caries is more than 2 times lower than in the norm, and after the lecithin-calcium complex intake significantly increases, although it does not return to normal.
The results of oral dysbiosis degree calculations regarding the relative activities of urease and lysozyme are shown in the Fig. 1, from which it can be seen that in persons with dental caries the degree of dysbiosis is 3 times higher than normal, and after a course of lecithin-calcium treatment it decreases almost by 8 times. This indicates the ability of the lecithin-calcium complex to regulate oral microbiocenosis, which certainly has a beneficial effect not only on the teeth condition, but also on other tissues of the oral cavity, and possibly on the whole organism state.
The change in the API index in patients with caries before and after the lecithin-calcium complex treatment is shown in the Fig. 2.
From these data it is clear that the index of API falls in patients by 2 times, and after treatment returns to normal. Since this index reflects the state of one of the body's defense systems, then, the lecithin-calcium complex in the form of the drug lecithin-calcium complex ensures the body's defense systems restoration.

Conclusions
1. In patients with multiple dental caries, the oral cavity dysbiosis, decrease in the protective antioxidant system level and decrease in the oral fluid calcium concentration are observed. Control group (1), patients with multiple caries (2), and patients after the lecithincalcium complex treatment (3). Control group (1), patients with multiple caries (2), and in patients after the lecithin-calcium complex treatment (3).