RU2708090C2 - Method for assessing level of mineralizing potential of oral fluid in children - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: invention relates to the field of medicine and represents a diagnostic technique for the risk of developing and exacerbating the activity of the decay process, comprising assessing the level of the mineralizing potential of the oral fluid, for which human saliva is centrifuged for 10 minutes at 3000 rpm with subsequent application of three supernatant drops onto chemically pure glass and temperature control at t = 37 °C for 30–40 minutes for examination under a microscope and photographing for 18–36 hours with subsequent use of an image analysis program "Image Tool 2.0" in order to determine distance between parallel tree-like crystals of samples with subsequent calculation of average value for determining level of mineralizing potential of oral fluid, which can be favourable in type of microcrystallisation with digital expression of 1–127.0 pixels and unfavourable at 127.1 pixels and more; and an unfavourable level of the mineralizing potential of the oral fluid, a high risk of developing and aggravating the activity of the caries process in children is diagnosed, while a favourable risk shows a low risk.

EFFECT: invention provides higher accuracy and diagnostic objectification.

1 cl, 3 dwg, 2 tbl

Description

The invention relates to the field of medicine, namely pediatric dentistry and can be used for scientific research and in the practical work of a dentist to quantify the level of mineralizing potential of the oral fluid (UMPS) in order to determine the individual risk of development and aggravation of the activity of the carious process in children.

The condition of the organs of the oral cavity to a large extent depends on the composition and properties of the surrounding fluid, namely the oral fluid.

Oral fluid plays a crucial role in maintaining the physiological balance of the processes of remineralization and demineralization in tooth enamel, especially in childhood. The mineralization and demineralization of tooth enamel depends on the mineralizing potential of the oral fluid, which is associated with the concentration of calcium ions and phosphate ions in the oral fluid at a certain pH of the medium. The active ion exchange between tooth enamel and oral fluid is also facilitated by the micellar structure of saliva, which maintains the mineralizing potential of the medium in an optimal state (Leontyev V.K., 2007). Visually, the level of mineralizing potential of the oral fluid can be assessed by determining the type of microcrystallization of the oral fluid.

A qualitative study of microcrystallization of saliva (ISS) in cariocontrolled individuals was carried out by Leus P.A. (1977). With multiple caries, erosion, and necrosis, he described three types of microcrystallization: type I - a clear picture of elongated crystal-prismatic structures fused together and occupying the entire surface of the drop; Type II - in the center of the droplet, individual dendritic crystal-prismatic structures of smaller sizes are visible than with type I; Type III - a large number of isometrically located irregular crystalline structures are visible throughout the drop. It has been clinically proven (Skripkina G.I., 2012) that the optimal for enamel mineralization in children is I and II types of ISS, which characterize a low level of risk of tooth caries.

To date, the description of the morphology of crystals is reduced mainly to a qualitative assessment. This is inconvenient due to subjectivity in data processing. There are attempts to translate a qualitative assessment of the ISS type into a quantitative one. Thus, a number of authors (Denisov A. B., 2007, 2011) collected a base of morphological features of the ISS, systematized and created their own morphological criteria for describing the quantitative characteristics of the types of microcrystallization of the oral fluid in healthy people. They developed an algorithm for evaluating crystal figures. In their opinion, 4 types of crystallization are normal in mixed saliva. A classification of morphological characters of crystalline aggregates has been developed, which reflects quantitative and qualitative characters (a total of 16 characters). Accordingly, the authors recorded changes in the crystallization of oral fluid in pathological conditions from various body systems. The cumbersome methodological approach presented is not without drawbacks related to the complexity, subjectivity and laboriousness of implementation, which imposes a limitation on its clinical use.

A known method for diagnosing the effect of a video display terminal (RF patent 2311639) [7] on the condition of the oral cavity, in particular on the condition of the teeth, and in particular to a method for determining the level of exposure to computer radiation on the condition of the teeth. According to the method, the pattern of microcrystallization of saliva (ISS) of a person associated with work on a computer is qualitatively determined, by the nature of which the level of influence of computer radiation on the condition of the teeth of the subject is evaluated. There are 5 types of ISS, which determine a different level of exposure to computer radiation on the condition of the teeth. The disadvantage of this method is the impossibility of an objective (digital) assessment of the level of mineralizing potential of the oral fluid, as well as the duration and complexity of the proposed method for determining the concentration of 11-hydroxycorticosteroids in saliva, which requires special equipment and reagents, the need for qualified chemists for analysis.

The technical problem solved by this invention is to increase the accuracy and objectivity of diagnosing the level of mineralizing potential of the oral fluid (UMPS) by quantitatively evaluating the type of ISS and expanding the possibilities for diagnosing the risk of developing and exacerbating the degree of activity of the carious process in the clinic of pediatric dentistry.

The proposed method was carried out as follows.

The research material is human saliva, which is taken in the morning, on an empty stomach, before brushing your teeth in a sterile test tube with a tight-fitting lid. The collected saliva is centrifuged for 10 minutes at a speed of 3000 rpm in a laboratory medical centrifuge OPL-8.

Next, a supernatant of centrifuged saliva from a test tube is taken with a sterile pipette for blood sampling and three drops are applied to chemically clean glass previously treated with alcohol and defatted with ether. Then the glass is placed in a thermostat at t = 37 ° C for 30-40 minutes. After time, the drug is examined under a microscope. From each drug, that drop is selected, the microcrystallization pattern of which occurs at least two times. Within 18-36 hours they are photographed.

Microphotography is carried out under standard conditions using an MBI-1 microscope with a Panasonic Lumix digital camera. To quantify the microcrystalline structure and measure the distance between the trunks of dendrites, the image analysis program “Image Tool 2.0” is used. This program allows you to open the profile of the optical density of the image. Black-and-white grayscale photographs in TIFF format are analyzed. In each picture, the region with the most pronounced quasiperiodic structure (4-5 dendrites) is selected and the distance between each dendrites is calculated using this program (Fig. 1, 2, 3).

(Fig. 1. Determination of the distance between dendrites with type I ISS.)

(Fig. 2. Determination of the distance between dendrites with type II ISS.)

(Fig. 3. Determination of the distance between dendrites in type III of the ISS.)

The technique of the work in the program:

1) Open the Image Tool

2) Open the picture “File-Open Image”

3) All drawings are converted to black and white (“Processing-Color-to-Grayscale”)

4) The size of each picture is reduced to 1: 2

5) By pressing the “Line Profile” button, the distance between the trunks of dendrites is measured;

6) The distance is determined in the program “length = ... pixels”

7) The arithmetic average for each type of microcrystallization of saliva is calculated.

During crystallization of samples, parallel tree-like crystals are formed - dendrites. Such dendrites form a quasiperiodic structure. The main difference between quasiperiodic structures for different samples is the difference in the distances between the stems of the dendrites. The indicated distances serve as a quantitative parameter characterizing the difference in the crystal structures of different samples. This parameter can be called the periodicity parameter. Samples with a clear visual difference have a different periodicity parameter.

For each type of microcrystallization (I; II; III) of saliva, 30 distances between the stems of the dendrites were chosen, with a clear visual difference. In the micrograph, a region with a quasiperiodic structure (4-5 dendrites), characteristic of this sample, was determined. Using the program “Image Tool 2.0”, the distances between the stems of dendrites (pc) for each type of microcrystallization of saliva were calculated.

 Next, we calculated for each distance in Excel.

 The data obtained were averaged for each group of samples. Next, the error of the periodicity parameter was calculated using the standard method of statistical processing of the results of direct measurements using Student's criterion.

The results of measuring the periodicity parameter are shown in table 1.

Table 1

The results of measuring the periodicity parameter for various types of ISS

ISS TYPES I
(pc) II
(pc) III
(pc) L1 44.8 58.92 98.24 L2 48.33 144.06 125.3 L3 43.86 76.84 68 L4 57.27 120.88 117.71 L5 42.19 137.78 78 L6 61.19 83.15 56.64 L7 116.52 144 114.02 L8 152 82.97 84.31 L9 110.02 124 80 93.76 188 165.47 107.52 76.16 68.21 108.83 52.8 80 132.24 91.3 114.02 148.61 58.14 95.41 154.95 62.61 116.36 70.71 98 117.05 69.23 100.58 112.87 85.44 34.93 110.35 90.38 58 224.4 117.05 72 88.84 130.97 60.73 104.84 99.48 125.87 130.6 66.48 100.06 67.94 115.13 102.98 91.61 160.46 88.57 261.2 98.68 97.67 240.61 70.77 110.08 158.35 75.07 65.79 228.58 117.1 110.45 178.57 116.82 68.03 235.55 M ± m 96.86 ± 5.67 93.18 ± 13.92 127.1 ± 23.98

Note: L _i - periodicity parameter;

                      pc - pixels

The results of the studies revealed statistically significant differences in the periodicity parameters between types I and II, between types II and III (p <0.001).

Thus, the proposed methodology for determining the periodicity parameter characterizing the quantitative difference in the types of ISS of saliva can be used to assess the level of mineralizing potential of the oral fluid (UMPS) in order to diagnose the risk of development and aggravation of the activity of the carious process in childhood.

The specified technical result is achieved by the fact that the determination of the distances between the trunks of dendrites (pc) using the program "Image Tool 2.0" and calculating the average for each sample.

Significant distinguishing features of the proposed technical solution are that the assessment of the level of mineralizing potential of the oral fluid (UMPS) (a favorable level of the mineralizing potential of the oral fluid, an unfavorable level of the mineralizing potential of the oral fluid) is carried out by quantitative determination (PKS) of the type of ISS saliva that does not require special equipment , reagents and qualified chemists for analysis to diagnose the risk of development and exacerbate activity ty carious process in childhood.

The proposed method has been clinically tested at the Department of Pediatric Dentistry, Omsk State Medical University during examination and dynamic monitoring of 60 caries-resistant patients from 12 to 15 years. ISS studies were carried out every six months for two years. After the observation period, in 89% of patients with type III of the ISS, dental caries was diagnosed, and in patients with type I and II of the ISS, caries was not detected.

Comparing the data of laboratory research and the results of long-term clinical observation of caries-resistant children, we can draw the following conclusion. A favorable level of mineralizing potential of the oral fluid is the type of ISS with a digital expression of 1 - 127.0 pixels; an unfavorable level of the mineralizing potential of the oral fluid is the type of the ISS with a digital expression of 127.1 pc or more.

The testing showed that this invention allows to solve the technical problem: improving the accuracy and objectivity of diagnosing the level of mineralizing potential of the oral fluid (UMPS) by quantitatively evaluating the type of ISS and expanding the possibilities of diagnosing the risk of development and aggravating the degree of activity of the carious process in the clinic of pediatric dentistry

The inventive method is confirmed by clinical examples.

table 2

Clinical examples

The patients one 2 3 4 5 ISS (pcs) 70.71 95.18 178.57 235.76 93.11 The level of mineralizing potential of the oral fluid (UMPS) favorable favorable adverse adverse favorable The risk of caries Low risk of caries Low risk of caries High risk of caries High risk of caries Low risk of caries Clinical Observation Results No caries growth after 2 years No caries growth after 2 years The increase in caries after 1 year. The increase in caries after 8 months. No caries growth after 2 years

Literature:

1. Denisov A. B. Saliva and salivary glands / A. B. Denisov. - M.: Publishing House RAMS, 2006. - 372 p.

2. Leontyev V. K. The study of saliva in dentistry: a method. recommendations / V.K. Leontiev, V.G.Suntsov. - Omsk, 1974.- 15 p.

3. Leontiev V. K. Caries and mineralization processes. (Development of methodological approaches, molecular mechanisms, pathogenetic substantiation of the principles of prevention and treatment): dis. ... Dr. honey. Sciences / V.K. Leontyev; MMSI. - M., 1978. - 541 p.

4. Leontiev V. K. On the micellar state of saliva / V. K. Leontiev, M. V. Galiulina // Dentistry. - 1991. - No. 5. - S. 17-20.

5. Leontiev V. K. Prevention of dental diseases / V. K. Leontiev, G. N. Pakhomov. - M., 2006 .-- 416 p.

6. Skripkina G.I. Prenosological diagnosis and prediction of the carious process in children (clinical and laboratory research, mathematical modeling) / G.I. Skripkina / Diss ... MD - Omsk, 2012 - 442 s.

7. Belskaya L.V., Golovanova O.A., Lomiashvili L.M., Borisenko M.A. A method for determining the level of exposure to computer radiation on the condition of the teeth. RF patent No. 2311639 dated March 31, 2006.

Claims (1)

A method for diagnosing the risk of development and aggravation of the activity of the carious process, which consists in assessing the level of mineralizing potential of the oral fluid, for which human saliva is centrifuged for 10 minutes at a speed of 3000 rpm, followed by applying three drops of the supernatant to chemically clean glass and thermostating at t = 37 ° C for 30-40 minutes for examination under a microscope and photographing for 18-36 hours, followed by the use of the Image Analysis 2.0 image analysis program to determine dividing the distance between parallel tree-like crystals of the samples, followed by calculation of the average value and comparison with control figures to determine the level of mineralizing potential of the oral fluid, which can be favorable for the type of microcrystallization of saliva with a digital expression of 1-127.0 pixels and unfavorable for the type of microcrystallization of saliva with digital expression of 127.1 pixels or more; and with an unfavorable level of mineralizing potential of the oral fluid, they diagnose a high risk of development and aggravation of the activity of the carious process in children, and with a favorable level of the mineralizing potential of the oral fluid - low risk.

Images