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10SIgA, in more than 70% of the subjects, which affected the average values of the indices of local immunity.
Before the start of treatment of patients with exacerbated course of chronic catarrhal gingivitis, an increase in the content of immunoglobulins of all classes (sIgA, IgG, IgM) in saliva was noted (Table 2). The im-munocorrection with lycopid at the 2nd stage of complex treatment of patients with generalized catarrhal gingivitis indicated a favorable effect on the clinical condition of the gums and restoration of local humoral protection of the oral mucosa in 100% of patients.
In the 2nd group of patients with exacerbated chronic catarrhal gingivitis immediately after treatment, 6-12 months after the treatment, stable normalization of sIgA, IgG, IgM was observed, which indicated the relief of inflammation processes and the absence of relapses of the disease in 96.3% of cases.
Conclusions
1. At the first stage of treatment of exacerbated chronic generalized gingivitis, it is recommended to carry out, along with rational professional measures, local antibiotic therapy with chlorhexidine-containing agents in combination with biosporin. Under their influence, there was a decrease and complete disappearance of potential pathogens of the disease and the normalization of the biocenosis of the gingival tissue in 93% of the treated.
2. The proposed scheme of immunotherapy for exacerbated chronic catarrhal gingivitis is recommended to be carried out after elimination of active inflammation in the gingival tissue. The use of lycopid at a dose of 1 mg per day for 10 days orally at the 2nd stage of treatment and subsequently for prophylactic purposes
(locally at other stages of complex therapy) makes it possible to accelerate the elimination of the main symptoms of the disease in 96.3% immediately after im-munocorrection and persistent clinical laboratory results throughout the entire period of dispensary observation in 100% of patients.
References
1. Belousov NN. Prichiny shirokogo raspros-traneniya tyazhelykh form vospalitel'nykh zabolevaniya parodonta. Parodontologiya. 2005; 3(36): 26-29.
2. Tsepov LM, Nikolayev AI, Mikheyeva YeA. Diagnostika, lecheniye i profilaktika zabolevaniy parodonta. M.: MedPress. 2008:272.
3. Hujoel P, Zina L, Cunha-Cruz J, Lopez R. Specific infections as the etiology of destructive periodontal disease: a systematic review. J. Oral Sci. 2013;121:2-6.
4. Rescala B, Rescala B, Rosalem W, Teles RP, Fischer RG, et all. Immunological and microbiologic profiles of chronic and aggressive periodontitis subjects. J. Periodontol.2010;81:P.1308-1316.
5. Cochran DL. Inflammation and bone loss in periodontal disease.J. Periodontal. 2008;79:1569-1576.
6. Michael PM. Immunological and Inflammatory Aspects of Periodontal Disease. Continuing Education Course. 2013;2:1-18.
7. Manchini G, Garbonara AO, Heremans SF. Immunochemical quantitation of antigens by single radial immunodiffusion. Immunochemistry.1965; 6(2):234-235.
1МУНОЛОГ1ЧНА ТА МШРОБЮЛОГ1ЧНА ХАРАКТЕРИСТИКА ГЕНЕРАЛ1ЗОВАНОГО
КАТАРАЛЬНОГО Г1НГ1В1ТУ
Шостенко А.А.
ДВНЗ «Буковинський державний медичний унгверситет», кафедра стоматологи дитячого в1ку, ассистент
IMMUNOLOGICAL AND MICROBIOLOGICAL CHARACTERISTICS OF GENERALIZED
CATARRHAL GINGIVITIS
Shostenko A.
HSEE "Bukovinian state medical university", department of pediatric dentistry, assistant
Анотащя
У данш статп наведен отримаш результата дослвдження показнишв мюцевого ÎMyHrreiy та мжроф-лори порожнини рота у 42 хворих на генералiзований катаральний пнпвгт з хрошчним та загостреним перебп-ом.
Для щентифшацд основних пародонтопатогешв застосовували полiмеразнy ланцюгову реакщю з ви-користанням ДНК-зондiв iз зворотно1 ДНК-пбридизацп.. Бiохiмiчнy щентифжацш анаеробних бактерш, стрептокошв i грамнегативних бактерш проводили за допомогою тест-системи фiрми АР1 (Францiя).
Концентрацiю sIgA, Ig G i M визначали методом радiальноï iмyнодифyзiï в гелi по G. Manchini.
Встановлено, що провiдна роль у розвитку захворювання належить порушенню стану мiсцевого iмy-нiтетy. Резидентна умовно - патогенна стрептококова мiкрофлора займае провiдне мюце в етiологiï хронь чного запального процесу в тканинi ясен. У розвитку загострень - домiнyючy участь приймае стафшоко-кава та грибкова iнфекцiï в рiзних асоцiативних взаеминах з гемолгтичними стафiлококами.
Abstract
This article presents the results of a study of local immunity and oral microflora in 42 patients with generalized catarrhal gingivitis with chronic and acute course.
To identify the main periodontal pathogens used polymerase chain reaction using DNA probes from reverse DNA hybridization. Biochemical identification of anaerobic bacteria, streptococci and gram-negative bacteria was performed using a test system from ARI (France).
The concentration of sIgA, Ig G and M was determined by the method of radial immunodiffusion in a gel according to G. Manchini.
It has been established that the leading role in the development of the disease belongs to the violation of local immunity. Resident conditionally - pathogenic streptococcal microflora occupies a leading place in the etiology of chronic inflammatory process in the gum tissue. Staphylococcal and fungal infections play a dominant role in the development of exacerbations in various associative relationships with hemolytic staphylococci.
Ключовi слова: катаральний пнпвгг, мшрофлора порожнини рота, мюцевий iMyHrreT, запалення.
Keywords: catarrhal gingivitis, oral microflora, local immunity, inflammation.
Catarrhal gingivitis is one of the most common gum diseases in the young population of industrialized countries. The results of many studies indicate a very unfavorable effect of chronic infectious-inflammatory process in the soft tissues of the periodontium on health and the growing accumulation of risk factors causing the transition of catarrhal gingivitis to a more severe and reversible disease - generalized periodontitis [1]. Based on this, effective rehabilitation of patients with chronic generalized gingivitis is an urgent preventive measure for the healing of periodontal tissues, as up to 50% of patients lose teeth in adulthood due to the progression of the inflammatory-destructive process in the periodontium.
However, to date, there is no optimal organization of treatment and rehabilitation of patients with generalized catarrhal gingivitis, depending on the features of its clinical manifestations, which is due to the diversity of etiological and pathogenetic mechanisms of their development. Given the above, in the development of treatment and rehabilitation measures should seek to take into account the most important factors causing the emergence of chronic and exacerbated variants of the disease and thus get a good idea of the etiological and pathogenetic nature of the disease, only on this basis.
[5.8].
A very important practical aspect is the establishment of dominant bacterial pathogens of exacerbated and chronic inflammatory process in the gingival tissues of patients with generalized catarrhal gingivitis
[7.9]. In addition to the leading etiological process (in the form of the causative agent) of various variants of the clinical manifestation of the disease, a significant role may belong to the state of local immunity, which provides primarily biocidality of the oral mucosa [4].
To date, the immune mechanisms involved in the formation of clinical variants of generalized catarrhal gingivitis have been insufficiently studied. Most authors (II Samoilenko, 2002, AA Gudaryan, 2005, GF Beloklitskaya, YG Chumakova, 2008, etc.) consider shifts in catarrhal gingivitis as primary, ie arising in preclinical stage of the disease, however, it cannot be ruled out that with a long course of catarrhal gingivitis, secondary immune disorders may develop, which develop in response to a constant inflammatory focus in the gingival tissues. In this regard, it is of particular interest to study the features of local (local) immunity, which would clarify the pathogenesis of both chronic
and exacerbated course of generalized catarrhal gingivitis.
Insufficient informativeness of generally accepted clinical indicators of the severity of inflammation in gingival tissues prompted us to search for simple and accessible laboratory criteria that could be used in the diagnosis of chronic and exacerbated generalized ca-tarrhal gingivitis.
It is known that in the acute phase of the inflammatory process, the production of so-called acute-phase reactant proteins, in particular alpha-2r protein, increases in body fluids. The study of fluctuations in mixed saliva will undoubtedly contribute to a more accurate and objective assessment of the manifestations of the disease.
Materials and research methods. We carried out clinical, laboratory and bacterial examination of 42 patients with generalized catarrhal gingivitis aged 19 to 31 years (mean age 25.6 ± 1.2 years). Among the examined were women - 23 (54.7%), men - 19 (45.3%).
The diagnosis of chronic and aggravated course of catarrhal gingivitis was carried out in accordance with the classification of inflammatory diseases adopted in Ukraine.
The present study included patients with a confirmed clinical and radiological diagnosis of generalized catarrhal gingivitis, without concomitant inflammatory diseases and chronic foci of infection in the periapical region.
The control group consisted of practically healthy donors - volunteers (18 people). Informed consent to research was required.
All patients underwent standard clinical studies, including collection of complaints, anamnesis, determination of allergological status, visual and instrumental assessment of the condition of the gum tissue.
To determine the hygienic state of the oral cavity, the Green-Vermillion method was used, the activity of the inflammatory process - the Mullemann bleeding index in Cowell's modifications [3].
Digital standard orthopantomography was used in the work for a more objective assessment of the diagnosis of the state of the bone tissue of the interdental alveolar processes.
To assess the activity of the inflammatory process in patients with generalized catarrhal gingivitis in mixed unstimulated saliva, alpha-2r glycoproteins were
determined by immunodifuse titration in agar gel according to Ounterloni using standard test systems according to the developers' method.
To determine the qualitative composition of bacterial agents, the method was used clinical bacteriological research, as well as the express method of polymer-ase reaction (PCR) followed by reverse DNA hybridization.
Microbiological studies were carried out on the material taken from the periodontal groove, after rinsing the oral cavity with distilled water.
Mixed unstimulated saliva, which was taken between 8-9 o'clock in the morning, strictly on an empty stomach, was subjected to special studies. During the work, the equipment, methods and reagents used did not change.
To assess the state of local immunity of the oral cavity, the levels of secretory immunoglobulin A (sIgA) were determined.
The concentration of sIgA, Ig G and M was determined by the method of radial immunodifusion in a gel according to G. Manchini [8]. Used monospecific serum against the named immunoglobulins.
We used traditional statistical methods using the Statistica for Windows 7.0 software package on a personal computer. The data obtained are presented as the arithmetic mean and standard deviation of errors. To assess the reliability between the groups, t - Student's test was calculated. At p< 0.05, the differences were considered significant [9].
Research results and their discussion. Analyzing the clinical data obtained, it should be considered that in selected patients with generalized catarrhal gingivitis, the manifestation of the disease was ambiguous in 24 (57.1%) patients according to clinical criteria (severity of pain, hyperemia, edema), bleeding index, the pathological process proceeded as chronic inflammation; in others, an exacerbation of inflammation in the gingival tissue was recorded.
Our study of the content of alpha-2r glycoprotein in the capillary blood of the inflamed gums showed that in the overwhelming number of patients - 21 (50%) chronic generalized gingivitis (group II) its content ranged in the range from 9.0 mg / l to 20.0 mg / l, and only in 5 (11.9%) concentration levels exceeded more than 24 mg / l, which indicated the highest activity of the disease manifestation. This served as the basis for the inclusion of the named patients for further studies in group I.
According to clinical and laboratory parameters, two groups of patients were formed for further studies. The first included patients with chronic manifestation of the disease (21 people); in the second with an aggravated course (21 pers.). Microbiological studies of the contents of the gingival groove showed that chronic and exacerbated generalized catarrhal gingivitis have differences in the etiological structure of possible causative agents of the above pathological processes.
Table 1
Microbiocenosis of the periodontal sulcus in patients with generalized catarrhal gingivitis with different clinical
course
Type of microorganisms Study groups
I group (n = 21) II group (n = 21) Healthy faces (n= 18)
a6c. % a6c. % a6c. %
Lactobacillus spp. 27 80,9 11 56,2 14 100
Str. salivarius 14 66,7 15 71,4 4 28,5
Str. sangvius 16 76,2 10 46,7 2 14,2
Str. haemoliticus 12 57,7 14 66,7 1 7,9
Str. epidermidis 13 61,9 11 56,2 0 0
Peptostreptococcus 11 52,3 4 28,5 1 7,8
Stf. aureus 2 9,5 11 56,2 1 7,8
Enterobacter 0 0 2 9.5 0 0
Bacteroides spp. 1 4,8 4 19,0 0 0
Fusobacteria spp. 1 4,8 3 14,3 0 0
Candida albicans 1 4,8 10 46,7 0 0
In positive cultures of material taken from patients with chronic generalized gingivitis, aerobic microflora was mainly detected, among which conditionally pathogenic microorganisms were of priority importance: S. Salivarius (66.7% of cases), S. Epidermidis (61.9% of cases), S. Haemoliticus (57.7% of cases), Peptostrepto-coccus (52.3% of cases). The listed bacteria were most often detected in 2-3 component associations.
In patients with an aggravated course of generalized catarrhal gingivitis, the microbial spectrum expanded due to the appearance of staphylococci and Candida fungi in the gingival tissues in a larger number of patients (Table 1).
With the data presented in Table 1, it can be seen that the biocenosis of the periodontal sulcus in patients with chronic generalized catarrhal gingivitis is characterized by a wide range of aerobic bacteria. The central place in the development of exacerbations belongs to staphylococci and fungi of the genus Candida.
Further studies have revealed the dependence of dysbiotic disorders in the gingival tissues and the state of local immunity.
When analyzing the levels of the main immuno-globulins in the oral fluid (saliva), in patients of both groups, the most indicative are the results of the content of sIg A and immunoglobulins of class G and M. A significant increase in the level of Ig G and a decrease in
SIgA parameters were revealed, which indicates a defi- both patients with chronic and exacerbated course of ciency of antibacterial protection of the oral mucosa in the disease (Table 2).
Table 2
Indices of local immunity in patients with generalized catarrhal gingivitis with various manifestations of the disease.
Immunity Indicators Study groups Normative Values (n = 18)
The first group of patients (n = 21) Second group of patients (n = 21)
M ± m M ± m
sIg A (г/л) 0,74±0,03* 0,34±0,02* 1,24±0,23
Ig G (г/л) 0,82±0,02* 0,60±0,02* 0,47±0,02
Ig M (г/л) 0,41±0,03* 0,28±0,03* 0,23±0,01
Note: * - p< 0.05 significant in relation to the indices of healthy people;
In patients with an exacerbated course of generalized catarrhal gingivitis, a significantly greater decrease in sIg A and Ig M levels and a lower increase in Ig G were revealed as compared with patients, having a chronic manifestation of the inflammatory process in the gingival tissues. The absence of an increase in the increase in Ig G and Ig M, as well as a different inhibition of sIg A production, indicates an inadequacy of the local immune response to a bacterial infection and may be one of the factors causing the appearance of staphy-lococcal and fungal infections in this category of patients (Table 2).
Conclusions.
1. Conditionally pathogenic streptococcal microflora occupies a leading place in the etiology of chronic generalized catarrhal gingivitis. In the formation of exacerbations of inflammatory phenomena in the gingival tissue, the dominant role belongs to staphylococci and fungi of the genus Candida.
2. In patients with an exacerbated course of generalized catarrhal gingivitis, more significant differences were revealed in the indices of local humoral immunity compared with the group of patients with chronic manifestation of the disease, who were characterized by a large decrease in sIg A and Ig M levels and, to a lesser extent, an increase in Ig G.
References
1. Belousov NN. [Causes of widespread severe inflammatory periodontal diseases.] Periodontology. 2005;36(3):26-29. Russian.
2. Lakin GF. [Biometrics: 4th ed.]. Moscow. Vy-shaya shkola, 1990;352. Russian.
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4. Rescala B, Rosalem W Jr, Teles RP, Fischer RG, Haffajee AD, Socransky SS, Gustafsson A, Figueredo CM. Immunological and microbiologic profiles of chronic and aggressive periodontitis subjects. J. Periodontal 2010;81:1308-16.
5. Cochran DL. Inflammation and bone loss in periodontal disease. J. Periodontal 2008;79:1569-76.
6. Manchini G, Garbonara AO, Heremans SF. Immunochemical quantitation of antigens by single radial immunodiffusion. Immunochemistry. 1965;2(6):234-5.
7. Marsh PD. Dental plaque: biological significance of a biofilm andcommunity life-style. J. Clin Periodontal 2005;32(Suppl. 6):7-15.
8. Michael PM. Immunological and Inflammatory Aspects of Periodontal Disease. Continuing Education Course 2013;2:1-18.
9. Hujoel P, Zina L, Cunha-Cruz J, Lopez R. Specific infections as the etiology of destructive periodon-tal disease: a systematic review. Eur. J. Oral Sci 2013;121:2-6.
