RU2581921C2 - Method of diagnosing diseases of periodontal tissues at different stages - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: invention refers to medicine, namely to dentistry, and can be used for diagnostics of diseases of periodontal tissues at different stages. For implementation of method saliva is examined, as value of inflammatory process, concentration of free oxyproline by spectrophotometric method is determined. If < 0.80 mg/l, absence of oral cavity tissues pathology is diagnosed; if 0.80-1.00 mg/l, acute catarrhal gingivitis is diagnosed; if 1.01-1.50 mg/l, chronic gingivitis is diagnosed; if > 1.50 mg/l, periodontitis is diagnosed.

EFFECT: use of method allows more precise, in short term and with available for all clinical laboratories equipment diagnosis of periodontal tissues diseases.

1 cl, 1 tbl, 8 ex

Description

The invention relates to medicine, namely to dentistry, and can be used to diagnose diseases of periodontal tissues at different stages.

Periodontal diseases are destructive. Most often, people get sick with gingivitis, periodontitis, periodontal disease, which vary in speed, aggressiveness and the depth of the course of destructive processes in periodontal tissues.

With gingivitis, superficial gum inflammation occurs without compromising the integrity of the joints of the teeth and gums. Periodontitis is an inflammatory disease that affects periodontal very aggressively (Barer G.M. Therapeutic dentistry: textbook: at 3 o’clock. - M .: GEOTAR-Media, 2009. - 4.2. - Periodontal disease. - 224 p.: 236 ill.).

Periodontitis can rapidly progress and in a short period of time (several years) lead to the loss of all or part of the teeth (Bezrukova A.P. Periodontology. - M., 1999. - 332 p.)

Gingival inflammation is characterized by the destruction of collagen, the main protein of connective tissue, which, without a doubt, leads to an increase in saliva levels of an amino acid such as hydroxyproline, which is unique to collagen (Vavilova T.P. Biochemistry of tissues and oral fluids. M .: GEOTAR-Media, 2008, p. 215)

However, in the special dental literature for the diagnosis of periodontal diseases, we were unable to find information on the use of the concentration of oxyproline in saliva as a marker of these diseases. It is known that the diagnosis of periodontal disease is carried out based on a subjective assessment of the state of periodontal disease by the patient himself, for example, bleeding gums during brushing or pain during eating. But this is an advanced stage of gingivitis.

A known method for the diagnosis of periodontal disease by determining the glycogen content on the surface of the gums (Schiller-Pisarev test). For this, the mucous membrane of the gums is stained with Lugol's solution. The color change in different areas after staining is judged on the presence of an inflammatory process.

There is also a method for evaluating collagen metabolism (Patent for invention No. 2214596; Podkovkin V.G., Bondarenko L.M., Vlasov M.Yu., Avvakumova N.P., Gribkova O.V.), related to clinical diagnostic research based on the determination of the content of free and protein-bound hydroxyproline in saliva in the dynamics of healing of gastroduodenal ulcers.

A method for evaluating collagen metabolism involves collecting biological fluid, isolating free and protein-bound hydroxyproline from it by extraction with ethyl alcohol and centrifugation, where free hydroxyproline is isolated from the supernatant. To isolate protein-bound oxyproline, barium hydroxide hydrolyzate is used, followed by ion-exchange chromatography. The concentration of oxyproline is determined colorimetrically at a wavelength of 540 nm in a 0.5 cm cuvette by adding Erlich reagent. As biological fluid, saliva of the examined person in the amount of 4 ml is used. Add 16 ml of ethyl alcohol to it, mix, centrifuged for 15 min at a speed of 3000 rpm. The isolation of protein-bound hydroxyproline is carried out on a chromatographic column with a diameter of 3 mm, a filling height of KRS-8p 6 mm ion exchange resin, with a rate of neutralization of 0.2 mm per minute. Oxyproline is eluted with 1 ml of 1N. hydrochloric acid solution, after which the hydrochloric eluate is neutralized with 5n. sodium hydroxide solution, while collagen decomposition and its biosynthesis are evaluated by increasing the concentration of free or protein-bound hydroxyproline, respectively.

A known method for the diagnosis of inflammatory diseases of periodontal tissues (Patent for the invention No. 23239509; Zhitkov M.Yu., Emilenko EA, etc.), the essence of which is in determining the content of calcium, phosphorus, alkaline phosphatase activity, total and tetra-resistant acid phosphatase in mixed unstimulated saliva. However, the method is more time-consuming and expensive, requiring special equipment.

The technical result of the proposed method is to increase the accuracy of diagnosis with minimal time by determining the concentration of free oxyproline in saliva, which is an objective assessment of the stage of the inflammatory process in the oral cavity, and is performed on equipment that is available in any clinical laboratory.

In our method, the concentration of free oxyproline in the saliva of patients who went to the dentist, including complaints of bleeding gums and tooth mobility, was determined spectrophotometrically using a modified C.S. method. Tetyanets, which is used to determine free oxyproline in serum (S. S. Tetyanets. Method for the determination of free oxyproline in serum. Laboratory case, 1985, N 1, p. 61-62).

10 minutes before the saliva was taken, the mouth was thoroughly rinsed with warm water to remove food debris; mechanical means for cleaning the oral cavity before the saliva was taken were not used. Studies have suggested that the determination of oxyproline in saliva may be a method for the diagnosis and prevention of periodontal tissue diseases.

For analysis, 0.25 ml of saliva was poured into test tubes, 1 ml of absolute ethanol was added, centrifuged for 10 minutes at 3000 rpm. All supernatant was transferred to volumetric centrifuge tubes. With stirring, a 0.25 ml solution of chloramine T in acetate buffer (1: 4) was added. 0.25 ml of isopropyl alcohol was added. After 4 min, 0.75 ml of Erlich reagent was poured. The total volume of the solution was adjusted with ethyl alcohol to 2.5 ml. After mixing, the tubes were closed with stoppers and thermostated at 60 ° C for 30 minutes. Then cooled to 20 ° C. An absorbance was determined on an SF 2000 spectrophotometer at a wavelength of 558 nm against a blank sample in which the biological fluid was replaced with water.

, где D - оптическая плотность раствора биологической жидкости.The oxyproline content in each sample was determined by the calibration line in coordinates

where D is the optical density of the biological fluid solution.

The problem is solved in that the method uses the value of an objective indicator of the concentration of free oxyproline in saliva, which allows you to diagnose the inflammatory process on an objective basis. The research results are shown in the table, where it is shown that the concentration of free hydroxyproline in the saliva of patients varies within the limits defined by us. For patients with healthy periodontal tissues, the corresponding concentration range of free hydroxyproline is <0.80 mg / l; patients with acute catarrhal gingivitis - 0.80-1.00 mg / l; patients with chronic gingivitis - 1.01-1.50 mg / l; patients with periodontitis> 1.50 mg / l.

Clinical examples.

1. Patient Ivannikov born in 1999 turned to the dentist for the purpose of professional oral hygiene.

Complaints: on the presence of plaque on the teeth.

Objectively: The mucous membrane of the marginal gum is pale pink, moderately moist.

A clinical examination of the oral cavity revealed a small amount of soft plaque on the lingual surface of the chewing teeth. The gingival connection is not broken. There are no changes on the orthopantomogram.

With the help of clinical indices (KPU 18%, GAME 0.4, RMA 0) an objective assessment of the condition of teeth and gums is carried out.

The results of the study of saliva:

Free hydroxyproline 0.65 (mg / L)

Conclusion: there are no signs of inflammation and destruction of the hard tissues of the teeth. Is healthy.

Diagnosis: K 03.66 Plaque.

2. Patient Nazarov born in 1983 turned to the dentist for the purpose of professional oral hygiene.

Complaints: bleeding gums that appear when brushing your teeth and biting off solid foods.

Objectively: The mucous membrane of the marginal gum is slightly hyperemic, pink, moderately moist.

A clinical examination of the oral cavity revealed soft plaque on all groups of teeth. When probing the periodontal sulcus, slight bleeding is noted. The gingival connection is not broken. There are no changes on the orthopantomogram.

Using clinical indices (KRU 28%, GAME 0.8, RMA 8) an objective assessment of the condition of teeth and gums is carried out.

The results of the study of saliva:

Free hydroxyproline - 0.93 (mg / l)

Conclusion: there are signs of inflammation without destruction of the hard tissues of the teeth.

Diagnosis K05.1 Acute catarrhal gingivitis.

3. Patient Namemenenko born in 1987 turned to the dentist for the purpose of professional oral hygiene.

Complaints: bleeding gums during brushing and biting off solid foods.

Objectively: the mucous membrane of the marginal gum is slightly hyperemic, moderately moist.

Clinical examination of the oral cavity revealed supragingival tartar on teeth 3.1, 4.1. When probing the periodontal sulcus, bleeding is noted. The gingival connection is not broken. There are no changes on the orthopantomogram.

With the help of clinical indices (KPU 25%, GAME 0.6, RMA 2) an objective assessment of the condition of teeth and gums is carried out.

The results of the study of saliva:

Free oxyproline - 1.23 (mg / l)

Conclusion: there are signs of inflammation without destruction of the hard tissues of the teeth.

Diagnosis K05.1 Chronic gingivitis.

4. Patient Martirosyan born in 1987 appealed to the dentist - hygienist.

Complaints: bleeding gums when brushing your teeth and biting off solid foods.

Objectively: the mucous membrane of the marginal gum is pale pink, slightly hyperemic, moderately moist.

A clinical examination of the oral cavity shows a small amount of pigmented plaque on the approximate surfaces of the teeth. When probing the periodontal sulcus, bleeding is determined. The gingival connection is not broken. There are no changes on the orthopantomogram.

With the help of clinical indices (KPU 39%, GAME 0, RMA 2) an objective assessment of the condition of teeth and gums is carried out.

The results of the study of saliva:

Free oxyproline - 1.01 (mg / l)

Conclusion: there are signs of inflammation without destruction of the hard tissues of the teeth.

Diagnosis K05.1 Chronic gingivitis.

5. Patient Ivonina born in 1974 appealed to the dentist-hygienist.

Complaints: aesthetic defect, plaque, halitosis.

Objectively: the mucous membrane of the marginal gum is cyanotic, edematous, moderately moist. Clinical examination of the oral cavity shows supragingival and subgingival tartar 3.3, 3.2, 3.1, 4.1, 4.2, 4.3., Pigmented plaque on all groups of teeth. When probing, periodontal pockets 3.5 mm deep, gum recession 1-2 mm. On the orthopantomogram, destruction of the cortical plate of the interdental septa is observed.

Using clinical indices (KPU 50%, GAME 1.3 RMA 0) an objective assessment of the condition of teeth and gums is carried out.

Free oxyproline - 1.77 (mg / l)

Conclusion: there are signs of inflammation

Diagnosis K05.3 Chronic periodontitis. Mild severity.

6. Patient Larchikova born in 1986 I went to the dentist-hygienist for professional hygiene.

Complaints: plaque, bad breath.

Objectively: the mucous membrane of the marginal gum is slightly hyperemic, swollen, moderately moist.

Clinical examination revealed supragingival and subgingival tartar on teeth 3.3, 3.2, 3.1, 4.1, 4.2, 4.3, pigmented plaque on all groups of teeth, periodontal pockets with a depth of 3.5 mm when probing. On the orthopantomogram, destruction of the cortical plate of the interdental septa is observed.

With the help of clinical indices (KPU 47%, GAME 0.8, RMA 5) an objective assessment of the condition of teeth and gums is carried out.

Free oxyproline - 2.70 (mg / l)

Conclusion: there are signs of inflammation

Diagnosis K05.3 Chronic periodontitis. Mild severity.

7. Patient Sobolev born in 1978 turned to the dentist for the purpose of professional oral hygiene. Complaints: for the presence of dental deposits, bad breath.

Objectively: the mucous membrane of the marginal gum is hyperemic, edematous, moderately moist.

Clinical examination of the oral cavity revealed supragingival and subgingival tartar in the area 3.3, 3.2, 3.1, 4.1, 4.2, 4.3, pigmented plaque on all groups of teeth. When probing the periodontal sulcus, bleeding is noted. Periodontal pockets 4-5 mm deep. On the orthopantomogram, destruction of the cortical plate, resorption of the vertices of the interdental septa by 1 / 3-1 / 2 of the height is observed.

With the help of clinical indices (KPU 50%, IGRU 1,2, RMA 12) an objective assessment of the condition of teeth and gums is carried out.

The results of the study of saliva:

Free hydroxyproline - 3.31 (mg / l)

Conclusion: there are signs of inflammation.

Diagnosis K05.3 Chronic periodontitis. Moderate severity.

8. The patient Bobyatinskaya I.Yu. Born in 1991 turned to the dentist with complaints of an aesthetic defect, the presence of plaque.

Clinical examination showed weakly hyperemic, moderately moistened mucous membrane of the marginal gum, soft plaque in the neck of all groups of teeth, supragingival calculus on the teeth 3.2, 3.1, 4.1, 4.2. When probing the gingival sulcus, bleeding of the gums is observed. The gingival connection is not broken. There are no changes on the orthopantomogram.

Diagnosis K05.1 Acute catarrhal gingivitis

Using clinical indices (KPU 62% GAME 1.8 RMA 3) an objective assessment of the condition of the teeth and gums.

The results of the study of saliva:

Free oxyproline - 1.00 (mg / l)

Conclusion: there are signs of inflammation without destruction of the hard tissues of the teeth.

Claims (1)

A method for the diagnosis of periodontal tissue diseases at different stages, including the study of saliva by a spectrophotometric method, characterized in that the concentration of free oxyproline is determined as an indicator of the inflammatory process and, if the values are <0.80 mg / l, the absence of an oral tissue pathology is diagnosed; with values of 0.80-1.00 mg / l, acute catarrhal gingivitis is diagnosed; with values of 1.01-1.50 mg / l, chronic gingivitis is diagnosed; at values> 1.50 mg / l, periodontitis is diagnosed.