SU1734021A1 - Method for determining sensitivity of oral cavity mucosa to stomatologic preparation - Google Patents

Abstract

The invention relates to medicine, namely to dentistry. The purpose of the invention is to improve the accuracy of the method by determining the sensitivity in the local area of the mucosa of the tooth-jaw segment to the dental preparation. The sensitivity of the tissue in the oral cavity to the preparations was determined on the basis of an assessment of the change in the ratio and functional activity of the gingival cell cells after exposure to the gingiva of this preparation. The gingival fluid was taken from the gingival groove before and after contact and the number of leukocytes was determined relative to the level of epithelial cells, and the activity of neutrophils was determined by their ability to phagocytosis and with an increase in the level of both of these indicators they concluded that the tissue in the area of the tooth was increased in the absence of an increase in the level of normal sensitivity of the tissue to the drug. The method allows to determine the sensitivity of the tissue to drugs locally, which is important for the selection of drugs in the treatment of a particular tooth and various inflammatory processes in the region of the tooth-jaw segment. The method is simple to perform, reliable and can be used in dental clinics. 2 tab. SP

Description

The invention relates to medicine, namely to dentistry, and can be used in immunology and allergology.

There is a method of determining the sensitivity of the oral mucosa to drugs. Its essence is that in washing liquids from the oral cavity collected before and after rinsing the mouth with an aqueous solution of a drug, the amount and activity of neutrophils in the reaction to inhibit the migration of leukocytes is determined and, in the case of a change in these indicators after exposure to the mucous membrane of the oral cavity state the presence of an allergic reaction to the drug

To collect the washings, rinse the anterior oral cavity with saline for 2 minutes. Then the oral cavity is rinsed with a solution of penicillin or norsulfazole for 2 minutes, and 15 minutes after exposure to the drug, wash again.

However, this method does not allow to determine the local sensitivity to the drug, which may become inadequate in the region of the unfolding pathological process, which causes its low accuracy. In addition, the method involves contact with the drug throughout the oral mucosa,

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which is of concern to the patient, especially if he has an allergic reaction to the drug. A number of drugs used in tooth filling, which may develop hypersensitivity, are insoluble in water (for example, eugenol), therefore, it is impossible to determine the sensitivity of tissue to them.

The aim of the invention is to improve the accuracy of the method by determining the sensitivity to the drugs in the local region of the tooth-jaw segment.

This goal is achieved due to the fact that the study of leukocytes is carried out in the gingival fluid obtained from the gingival groove, and the mucous membrane is contacted with the preparation applied to the filter paper disk for 5-10 minutes after contact with the level of epithelial cells. current and activity — according to the ability to phagocytosis, and with an increase in these indicators, an increased sensitivity is determined, and in the absence of changes, the normal sensitivity of the oral mucosa to the therapeutic pre- paratha

The positive effect of the proposed method is achieved by combining methods of irritation of the mucous membrane in the local gum area as a result of contact with the drug and the study of changes in the number and functional activity of the cells of the gingival groove. Using antibiotic-impregnated filter discs to determine the sensitivity of microbial strains. At the same time, the impregnated disk is placed on the layer of agar on which the microorganisms are sown: intracutaneous administration of allergens to determine the allergic reaction to the preparation (if there is such a reaction, the local hyperemia, compartment); A sublingual allergen test is dropped under the tongue and after 5-7 minutes, the presence of an allergic reaction is determined by the general condition of the patient (weakness, nausea, etc.). All preparations used in the listed samples are water soluble.

The contact time with the drug for 5-10 minutes is optimal for determining the sensitivity of the tissue to the drug. A contact that is less than 5 minutes in length is insufficient to change the migration of leukocytes into the gingival groove, and therefore, to determine sensitivity. Contact for more than 10 min makes no sense, since it does not lead to further changes in the indicators of the number and activity of leukocytes. The duration of the interval between contact and analysis of cellular material was taken 20-30 minutes, assuming that the maximum number of cells accumulates in the groove 20 minutes after exposure and begins to decrease again 10 minutes later.

PRI me R 1. Sick P.46 years old diagnosis; exacerbation of chronic pulpitis 3. It is necessary to determine the sensitivity of the tissue in a given tooth to the eugenol preparation, which is used in canal filling. For this, gingival fluid was taken from the gingival groove of this tooth using a device consisting of a needle with a blunt end, tightly connected to a can (such as a syringe tube, which has a blunt needle tip). The end of the needle was inserted into the groove and the gingival fluid was sucked in, which was immediately transferred to the well of the immunological plate containing 0.05 ml of Hanks solution, resuspended, and stored until a second sample was taken from the groove.

Then, to the gum of this tooth, a disk of filter paper 0.5 cm in diameter, pre-impregnated with the preparation solution and dried (the disk was impregnated with 12.5% eugenol solution in ethanol, dried, and kept in a closed box at room temperature). The disk was kept on the gum for 5 minutes, after which it was removed. 25 minutes after removing the disc from the gum from the gingival groove, the gingival fluid was again collected and transferred to another well of the plate containing 0.05 ml of Hanks solution, resuspended.

0.05 ml of 0.05% suspension of filtered Baker's yeast cells was added to suspensions of two cell samples (taken before and after the effect of the drug on the gum). The yeast was prepared as follows: fresh yeast was mixed with a physiological solution in a 1: 2 ratio, kept at 80 ° C for 60 minutes, cooled, 10% formalin was added and stored at 4-10 ° C; washed before use at least 6 times. Centrifuged at 200 g for 5 min. After this, 0.025 ml of fixative was added (1.3% glutaraldehyde and 6% formaldehyde in Hanks solution) and held for 10 minutes. Then the supernatant was spun out of the wells, and 0.05 ml of the paint solution (methyl green pyrornine) was poured into the sediment. A triacetate cellulose film coated with a tanned layer of gelatin was applied on the tablet, pressed tightly with a metal lid with a foam pad 0.5 cm thick. The sediment in the wells was resuspended carefully, then precipitated onto the film by centrifuging the tablet at 200 g for 5 min. The film was removed, the preparation was dried in a stream of warm air, then washed with tap water and dried again. A layer of Canadian or fir balsam in xylene was applied to the finished products and glued to the glass slide. The ratio of leukocytes and epithelial cells and the number of phagocytic neutrophils in both preparations were calculated. In this patient, before contacting the gum with a disc impregnated with eugenol, the values of the indicators were as follows: the ratio of leukocytes: epithelial cells was 58:42, the number of phagocytic neutrophils — 18%. After contact with the drug, the ratio of leukocytes: epithelial cells was 64/36, and the level of phagocytosis was 24%. Thus, the first indicator (the number of leukocytes in relation to neutrophils) increased by 11, and the second by 33%. Based on this, it was concluded that the sensitivity of this patient to the drug. The immediate results of treatment showed the following. The use of zinc-Eugenol paste for filling the canal led to a slight swelling of the soft tissues of the lower jaw on the left, smoothing the transitional fold in the area of the treated tooth, its painful bite (although the canal was sealed to the top of the root). All these symptoms indicate an increased sensitivity of the tissue in the area of the tooth to eugenol.

PRI mme R 2. Sick T., 23 years old, diagnosis: chronic fibrous pulpitis. Eugenol was used to seal the canal. The sensitivity of the tissue in this tooth to this drug was tested. The gingival fluid was taken from the gutter of the patient in the same way as in example 1. It was transferred to the well of the plate containing 0.05 ml of Hanks solution, resuspended. Then, to the gingiva of this tooth, a disk of filter paper impregnated with a solution of eugenol was put in the projection area of the apex of the root (the discs were prepared in the same way as in Example 1) and kept for 10 minutes. Then the disk was removed and after 30 minutes from the gingival groove of the same tooth, the gingival fluid was taken. The reaction of phagocytosis was set as in Example 1. The ratio of leukocytes, epithelial cells and the number of phagocytic neutrophils was counted. Before contact with eugenol, the ratio of leukocytes: epithelial cells was 64:36, the level of phagocytosis was 16%. After contact, the ratio of leukocytes: epithelial cells became 62:38, the level of phagocytosis

The remaining 16%. Conclusion - there is no sensitivity to the drug, which has been confirmed clinically, since there are no complaints after treatment with the use of zinc-eugenol paste. On external examination: configuration of the face,

regional lymph nodes, as well as the mucous membrane in the area of the medical tooth are not changed. Percussion is painless,

PRI me R 3. Sick K., 43 years old, sent by the periodontist for depulpation

4 teeth and resorcin-formalin method, mobility of II degree. When treating this tooth, a resorcin-formalin mixture was used. The sensitivity of the tissue in this tooth to this drug was tested. Using the device (as in examples 1 and 2), gingival fluid was taken from the gingival groove of this patient, transferred to the well of the tablet containing 0.05 ml of Hanks solution, and resuspended. Then, a disk of filter paper impregnated with a solution of resorcinformalin mixture was attached to the gingiva of this tooth in the projection area of the apex of the root (the discs were prepared as follows,

prepared a 1.56% aqueous solution of resorcinformalin mixture, soaked them with filter paper discs, dried and stored at room temperature), kept for 7 minutes, then the disc was removed and

After 20 minutes, the gingival trough of the same tooth took gingival fluid, transferred it to another well of the tablet containing 0.05 ml of Hanks solution, and was resuspended. The reaction of phagocytosis was set as in Example 1. The ratio of leukocytes to epithelial cells and the number of phagocytic neutrophils were calculated. Before contact with the resorcinformalin mixture, the ratio of leukocytes: epithelial cells was 56:44, the number of phagocytic neutrophils was 6%; after contact with the drug, the ratio of leukocytes, epithelial cells was 66:34, the level of phagocytosis was 14%, i.e. the first indicator increased by

12, and the second - by 130%. It was concluded that the patient was hypersensitive to the drug. The immediate results of treatment using resorcinformalin mixture showed the following. The patient noted pain when biting on the tooth. At external examination: the configuration of the face, regional lymph nodes, and mucous in the area of the treated tooth are not changed. Percussion of the tooth is painful. Like

The first two examples of channels were sealed strictly to the top of the root.

The proposed method makes it possible to more accurately determine the sensitivity of the tissue to the preparation in the region of a particular tooth than the known method, the accuracy increases 5.4 times.

The increased accuracy of the method is achieved by analyzing the cells of the gingival chute after exposure in the local area of the mucous membrane of the periodontal segment. But there is one more reason. In the proposed method, the cells of the gingival groove are analyzed, and in the prototype, cells are washed out of the oral cavity. Their viability is different. The viability of gut gut neutrophils is on average 72.2 and 2.6%, and washout from the oral cavity - 61.2 3.2%. It is clear that the greater the number of viable cells isolated, the higher the accuracy of any immunological reaction.

The method is simple in execution. It takes less time than the prototype (table 2).

In addition, since the method offers manipulations with small numbers of cells (tens and hundreds less than the prototype), it therefore requires a minimum amount of reagents and materials. In contrast to the prototype, the proposed method is more objective, since the quality and number of cells washed. studied by the prototype largely depends on the strength, ability and thoroughness of mouth rinsing by patients, and the taking of cells from the gingival groove, studied by the proposed method, is carried out by the doctor all the time in the same way.

The method is ready for practical use. It is designed for use in the laboratories of dental hospitals and clinics.

Claims (1)

Invention Formula A method for determining the sensitivity of the oral mucosa to a dental preparation, including counting the number of leukocytes in the oral cavity and detecting their activity before and after contact of the mucous membrane with a medicinal substance, in order to improve the accuracy of the method by determining the sensitivity in the local area -maxillary segment, the study of leukocytes is carried out in the gingival fluid obtained from the gingival groove, and the contacting of the mucous membrane is carried out with prepa filter paper on the disc for 5-10 min, and counting the number of leukocytes lead 20-30 min, after contact with respect to the level of epithelial cells, and the activity of leukocytes on the ability to phagocytosis and Preservation of the initial indices is defined as the sensitive oral mucosa to the preparation. Table 1 Table 2 Operations The proposed method Collection of material, min. Contact with the preparation, min. Time interval before re-collection of material, min. Re-collection of material, min. Cell washing, mines preparation operations Reactions, min Calculation of results, min Total min Desnev liquid 2 min. 10-15 20-30 2 o Preparation of yeast cell suspension, instillation for 20 minutes Phagocytosis (taking into account centrifugation, fixation) 20 At the same time the ratio of cells and phagocytic activity of neutrophils is 78-93 Prototype Flush 2 Mouthwash 2 15 2 thirty Capillary filling, chamber preparation 40 min RTML (incubation) 60 RTML-2x1 2 min. The number of cells with regard to the preparation of the Gor eva-159 camera.