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18
шдукованими порушеннями нормально функцюну-вання оргашзму та психогенними депресивними змь нами.
Висновки
В ходi нашого дослiдження вперше показанi етапи морфогенеза В1Л-асоцшованих виразкових змiн в тканинах пародонта та СО порожнини рота. Виявлено, що у па^енпв з хворобами тканин пародонта на тл ВШ-шфекцп порушуються процеси десквамацп етте-лiю, що е свщченням зниження захисноТ ролi СО порожнини рота. ^м того виникае компенсаторно-адаптивна пперкератотична перебудова ештелю, як попередник СНIД-iндикаторних станiв та прояв максимального напруження резервних можливостей оргашзму. Ц дат пщтверджуються результатами обчи-слення Ыдексу кератинiзацiТ, значення якого зростае з прогресуванням пародонтопатолопТ та основного за-хворювання.
Таким чином, можливост цитологiчноТ дiагностики стану тканин ротовоТ порожнини людей зi статусом В1Л дозволяе об'ективiзувати патологiчнi та функцю-нальнi змши СО порожнини рота в цтому та ясен зо-крема при ВШчнфекцп. Це дозволяе пiдвищити ефек-тивнiсть ранньоТ об'ективноТ дiагностики розвитку та прогресування патолопчних процесiв в тканинах пародонта.
Лдоратура
1. Живиця Д.Г. Виживання хворих на В1Л-1нфекц1ю протя-гом 3 рок1в проведения високоактивноТ антиретров1ру-
сноТ терапп / Д. Г. Живиця // 1нфекц1йн1 хвороби. - 2011.-№ 3. - С. 21-26.
2. Александрша Т.А. В УкраТн - концентрована еп1дем1я В1Л/СН|Ду з тенден^ею до генерал1зацп, у черз1 на отримання аитиретровiрусиоí терапп - 9037 ос1б: бес1да з головою ДержавноТ служби соцзахворювань / Т.А. Алексаидрiиа; записав О. Устшов // УкраТнський меди-чний часопис. - 2011.-№ 6. - С. 16-17.
3. Данилевський М.Ф. Захворювання слизовоТ оболонки порожнини рота / М.Ф.Данилевський, О.Ф.Неан, Ж.1.Рахжй ; За ред. М.Ф.Данилевського. - К.: Здоров'я.
- 1998. - 408с.
4. Быков В.Л. Гистология и эмбриология органов полости рта человека / Быков В.Л. - СПб.: «Специальная литература», 1998. - С.19-25.
5. Колесова Н.В. Особливост альтерацп i репаративноТ регенерацп епiтелiю ясен при геиералiзоваиому паро-донти тi та Тхня фармаколопчна корекцiя: автореф. дис. на здобуття наук. ступеня канд. мед. наук : спец. 14.01.22 «Стоматолопя» / Н.В.Колесова. - КиТв, 2002. -20 с.
6. Быков В.Л. Цитология и общая гистология (функцона-льная морфология клеток и тканей человека) / Быков В.Л. - СПб.: СОТИС, 1998. - 516с.
7. В.Г.Бургонский Лекция: СПИД в аспекте стоматологического приема / В.Г. Бургонский // Современня стоматология. - 2002.- № 4. - С.108-112.
8. Гилева О.С. Биохимия слюны, клиника и профилактика заболеваний слизистой оболочки полости рта в условиях производственного воздействия табака: автореф. дис. на здобуття наук. ступеня канд. мед. наук : спец. 14.01.22 «Стоматология» / О.С.Гилева. - Москва, 1988.
- 15с.
9. Салюк О.Д. Рання цитолопчна дiагностика пнпвггу. Автореф. дис. на здобуття наук. ступеня канд. мед. наук : спец. 14.01.22 «Стоматолопя» / О.Д.Салюк. - КиТв, 2004. - 20 с.
English version: THE ANALYSIS OF THE CYTOLOGICAL PARAMETERS OF THE ORAL MUCOUS MEMBRANE EPITHELIUM IN HIV-POSITIVE
5k
PATIENTS*
Ilenko N.V, Petrushanko T.O., Nikolenko D.E.
Higher State educational if Ukraine "Ukrainian Medical Dental Academy", Poltava
The problem of HIV/AIDS is extremely burning worldwide. Especially for Ukraine, where this epidemic is spreading steadily and has a tendency to generalization. The aim of our study was the diagnosis of the cytoiogicai changes in the oral mucous membrane (OMM) epithelium in HIV-positive patients. Materials and methods. 94 HIV-patients aged 23-49 were examined. They formed the studied group (I). 35 people with HIV status-negative aged 21-45 entered the control group (II). This research work was conducted during 2011 - 2012. After general subjective and objective examination of patients both HIV-positive and HIV-negative, there was performed mucous membrane cytosmear scraping of the cheek and smear-reprint from interdental papilla of each patient. To analyze the barrier function of OMM its degree of keratini-zation was revealed with the help of the keratinization index (KI). Results. During our study the stages of morphogenesis HIV-associated ulcerative changes in the periodontal tissues and OMM were firstly showed. We found that epithelial desquamation disrupts in patients with periodontal diseases and HIV-lnfection, which is a testament of the protective role reduce of OMM. In addition there is a compensatory-adaptive hyperkeratotnic alteration of the epithelium, as a precursor to AIDS-indicator conditions and display of maximum stress reserve of the organism. These data are confirmed by the results of the of keratinization index calculation. Its value increases along with the progression of the underlying disease and periodontal diseases.
Key words: HIV-infection, periodontal diseases, cytological picture, keratinization index
The problem of HIV/AIDS is extremely burning worldwide. Herewith Ukraine, together with Estonia and the Russian Federation, has the leadership in Eastern Europe in terms of growth in the number of HIV-infected
people [1]. According to the official statistics as 01.10.2011 115,275 HIV-positive patients had regular medical check-up in Ukraine. Every year the HIV-infection is diagnosed firstly on average in 20,000 times
* To cite this English version: Ilenko N.V., Petrushanko T.O., Nikolenko D.E. The analysis of the cytological parameters of the oral mucous membrane epithelium in HIV-positive patients//Problemy ekologii ta medytsyny. - 2013. - Vol 17, № 1-2. - P. 9 -7.
in our country. This epidemic is spreading steadily and has a tendency to generalization [2].
A detailed study of pathogenesis associated with HIV-infection and AIDS is an essential factor for improving the diagnostic and treatment strategies of HIV-positive people, for making their quality of life better, for increasing their social adaptation and reducing stigma and discrimination against people with HIV status. The cytological methods have a special place in the diagnostic algorithm of HIV-patients examination. As they allow non-aggressively in other words safely and in a short time to determine the nature of pathological changes occurring in tissues, the traits of morphological manifestations of diseases and the adaptation processes of the organism to the action of pathogenic agents.
The aim of our study was the diagnosis of the cytological changes in the oral mucous membrane (OMM) epithelium in HIV-positive patients.
Materials and methods
94 HIV-patients aged 23-49 were examined. They formed the studied group (I). 35 people with HIV statusnegative aged 21-45 entered the control group (II). They didn't have any peculiarities of household and employment anamnesis. Clinical and laboratory examination of patients was conducted in Poltava Regional Center for AIDS Prevention and Control, Poltava Regional Dental Out-Patient Hospital and the Department of Pathology of the biopsy-sectional course HMEI of Ukraine "UMSA" during 2011 - 2012.
After general subjective and objective examination of patients both HIV-positive and HIV-negative, there was performed mucous membrane cytosmear scraping of the cheek and smear-reprint from interdental papilla of each patient. Smears were prepared on sterile skim slide, dried and Giemsa stained. Obtained cytological smears were studied using Olympus BX41 light microscope with objectives * 10, * 20, * 40, * 100, and pictured with an Olympus S3040-A DUP digital camera. To analyze the barrier function of OMM its degree of keratinization was revealed with the help of the keratinization index (KI). The total number of epithelial cells seen in the microscope field of view was counted, then the number of keratinizing cells were multiplied by 100 and divided by the total number of cells. Reducing of KI shows a decline of mucous membrane protective function [3].
Results and discussions
Microscopic examination of mucous membrane smear scraping taken from the cheek of HIV-positive patients and Giemsa stained reported that the cellular composition of the material is presented by superficial and intermediate epithelial cells. Superficial epithelial cells had a hexangular or rectangular shape, low nuclear-cytoplasmic ratio, dark rounded or oval eccentric nucleus and basophilic cytoplasm (Fig. 1). Intermediate cells were different in shape, often polygonal, larger in size, had higher nuclear-cytoplasmic ratio due to an increased nuclear. The latter ones had a round or oval form. These cells also had evident perinuclear filamentary structures (Fig. 2). Rarely some smears contained rounded epithelial cells with weak basophilic cytoplasm and eccentric nucleus that correspond to parabasal cells (Fig. 3).
npoSAeMH eKOAorii Ta MejHUHHH
0 0 % .
□
Figure 1. Mucous membrane scraping the cheeks taken from an HIV-positive person. Giemsa stained. Enlarged x 1000: 1 - superficial cell; 2 - intermediate cell; 3 - single coccus
bacteria.
? .
El
Fig.2. Mucous membrane scraping the cheeks taken from an HIV-positive person. Giemsa stained. Enlarged x 1000: 1 - intermediate cell; 2 - colonies of microorganisms.
Fig.3. Mucous membrane scraping the cheeks taken from an HIV-positive person. Giemsa stained. Enlarged x 200: 1 - intermediate cells; 2 - parabasal cells.
Parabasal cells get into the smears either of the deep epithelial trauma during collection of material for cytological examination, or the presence of pathological process [4].
Unlike cheek epithelium smears-reprints from the gingivae of HIV-positive patients contain less intermediate cells, but more surface cells and horny scales. Obviously, these changes of cytological components are connected
with gingival functional features, like increased chewing workload. This causes the appearance of a large number of horny scales.
During our research we firstly diagnosed the morphogenesis of stages of micro flora damaging effects on the OMM in HIV-patients. On the first stage HIV enters the cytoplasm of epithelial cells, HIV viral particles are formed. At the place of penetration, cells of spinous and basal epithelial layers react with dystrophic and necrobi-otic changes. The surrounding cells are changing adap-tively with the development of cornea dystrophy - hy-perkeratoric process (Fig. 4).
0
Figure 4. Smear-reprint from the gingival surface of the gums HIV-patients. Giemsa stained. Enlarged x 1000: 1 - necrobiosis of parabasal cells; 2 - a phenomenon of hyperkeratosis.
Reactive changes of epithelial cells in the place of HIV penetration have an inflammatory-dystrophic nature and manifest themselves as hydropic and balloon degeneration and spongiosis later on in the future. These pathological processes are the signs of wrongdoing protein-water-electrolyte metabolism, accompanied by changes in colloid-osmotic pressure in the cell. Penetration of microorganisms, mainly Gram "+", in the section desmosome contacts leads to their destruction. The cells lose contact with each another and become rounded. The vesicles, forming inside the epithelium, quickly disclose with the formation of ulcer or erosive surface (Fig. 5, 6). At the local of lesion a significant number of white blood cells are determined. There are a large number of fungal flora, represented by Candida, As-pergillus and Penicillium, in the broken integrity areas of the epithelial sheet
Figure 5. Smear-reprint from gingival surface of the person with HIV-associated ulcerative periodontitis. Giemsa stained. Enlarged x 1000: 1 - severe erosion; 2 - bacterial flora.
In the oral cavity the described cytological changes are reflected clinically in the development of HIV-associated ulcerative gingivitis and periodontitis in the dynamics of the pathological process.
Figure 6. Smear-reprint from gingival surface of the person with
HIV-associated ulcerative periodontitis. Giemsa stained.
Enlarged x 1000: 1 - ulcerative defect; 2 - proliferation of basal and parabasal cells.
Our results are agreed with the data provided by national scientists. So Kolesova N.V. (2002) mentioned a number of specific morphological changes that occur in the gingival epithelium due to the development of generalized inflammatory and degenerative processes in periodontal tissues. These include the development of granular, hyaline-drop and hydropic degeneration, damage the desmosome contacts and uneven expansion of intercellular spaces, their infiltration by elements of micro flora, focal alteration and desquamation of individual cells. The author points to the deepening of pathological changes in gingival epithelium with the progression of periodontal diseases. Dystrophic, necrobiotic and focal necrotic changes are developing while a number of desquamated epithelial cells are increasing at the same time [5]. Our results showed the similar changes, but with decreasing of desquamation processes and developing hy-perkeratotric changes that we associate with HIV infection and immunodeficiency progression.
It is known that one of the major function of OMM as a whole and the gums, in particular, is protection, which is provided by the barrier role of epithelium. It is stipulated by a number of factors: large epithelial layer thickness, the presence of numerous intercellular contacts, bad permeable chemically and mechanically stable corneous layer, constant exfoliation of surface layers and their rapid recovery, production of antimicrobial compounds and continuous wetting it with saliva [4,6].
To characterize the epithelial barrier function, we used the keratinization index (KI), which reveals the degree of keratinization of the gingival interdental papillae epithelium and assesses the level of the protective function of OMM. In HIV-infected persons KI exceeded that in patients without HIV status almost in 1.8 times. It was revealed the reducing of KI in the control group by 20% compared to the estimated rate. In our opinion, it is due to the fact that these patients have inflammatory and in-flammatory-dystrophic changes in periodontal tissue in 82.86% of cases. The periodontal diseases in HIV-infected individuals have been diagnosed in 98.94% of cases. Obviously, the increasing of KI in HIV-patients by 40% compared with an estimated rate can be attributed
Проблеми екологц та медицини
to the development of adaptive hyperkeratonic changes Results of calculation of KI in HIV-positive and HIV-
of gingival epithelium in response to the impact of oral negative individuals are shown in Table 1. micro flora associated with HIV. It is agreed with obtained cytological picture.
Table 1
Changes of keratinization index at different condition of periodontal tissues of examined individuals
Parameter Studied group (I) Control group (II) P
KI 70,51 ±1,45 39,4±1,45 <0,001
KI in presence of inflammatory changes in the periodontal tissues 51,13±7,21 40,28±1,89 <0,05
KI in presence of inflammatory-dystrophic changes in the periodontal tissues 72,27±1,35 34,87±1,94 <0,001
Note: p - the probability of error according to Student's tables.
We analyzed the KI data according to the intensity of periodontal lesions. Thus the significant variation in the results was found in the presence of inflammatory processes in periodontal tissues in the control group. The results of KI in HIV-patients and people without HIV status in presence of inflammatory periodontal diseases differed unreliable. Whereas, in presence of inflammatory-
dystrophic changes in the periodontal tissues, KI in the experimental group exceeded the average parameter in control group in 2 times.
For a group of HIV-patients we conducted an analysis of the KI results according to severity of periodontal diseases. The relevant data are given in Table 2.
Table 2
Changes of keratinization index in HIV-patients with different severity of periodontal lesions
Parameter Value P
Index 1 Index 2 Index 3 Index 4 Index 5
1. KI in presence of gingivitis 51,13±7,21 < 0,05 < 0,05 < 0,05 < 0,01
2. KI in presence of general periodontitis initial stage 67,47±3,89 < 0,05 > 0,05 > 0,05 <0,001
3. KI in presence of general periodontitis I stage 66,57±1,8 < 0,05 > 0,05 > 0,05 <0,01
4. KI in presence of general periodontitis II stage 72,04±5,45 < 0,05 > 0,05 > 0,05 > 0,05
5. KI in presence of general periodontitis III stage 82,35±1,71 < 0,01 < 0,001 < 0,01 < 0,05
Note: p - the probability of error between given parameters according to Student's tables.
It was revealed the tendency to an increase in the in- Analysis of the KI results in HIV-patients conducted in
tensity of keratinization of gingival epithelium in the pro- accordance with the stage of the underlying disease. gression of destructive changes in periodontal tissues in These data are presented in Table 3. HIV-patients.
Table 3
Change the keratinization index of OMM at different stages of HIV infection
Parameter Value P
Index 1 Index 2 Index 3 Index 4
KI in patients with I stage of HIV-infection (index 1) 50,26±3,54 < 0,01 < 0,01 < 0,01
KI in patients with II stage of HIV-infection (index 2) 66,65±1,23 < 0,01 < 0,001 < 0,01
KI in patients with III stage of HIV-infection (index 3) 75,65±1,37 < 0,01 < 0,001 > 0,05
KI in patients with IV stage of HIV-infection (index 4) 77,4±2,34 < 0,01 < 0,01 > 0,05
Note: р - the probability of error according to Student's tables.
A steady tendency to increasing the KI with progression of HIV infection is typical. It indicates the development of hyperkeratonic processes in the gums.
According to the data by L.A.Dmytriyevoyi (2001), exfoliation squamous cells with microorganisms in the cytoplasm is one of the protective reactions of surface gums epithelium. Increasing the KI is regarded by us as an indicator of reducing physiological desquamation of the epithelium, and thus a violation of the protective functions of surface epithelium gums in HIV-patients.
It is necessary to consider the fact that hairy leu-kokeratosis, together with HIV-associated periodontal diseases belongs to a group of oral lesions that are often associated with HIV-infection according to WHO classification [7]. On the other hand, according to the results of native researchers [8], the precursor of leukoma mucous
membrane changes is the development of diffuse hy-perkeratonic_processes and chronic inflammation. These data can be compared with the results of our research and consider hyperkeratonic changes of OMM epithelium, as a precursor to the development of hairy leu-kokeratosis of the tongue and oral keratosis.
It is known that a state of increasing keratinization is seen by Salukis O.D. (2004) as a premorbid state that occurs in response to an inappropriate environment. It is closely related to the mechanisms of stress reserve of the organism and indicates failure in the physiological processes of maturation of epithelial cells and adaptive compensatory adjustment in the epithelial layer [9]. In our opinion, hyperkeratonic changes of OMM epithelium in HIV-infected people have a similar mechanism of development, but occur in response to the impact of HIV-
associated oral micro flora, together with somatic immune changes, stress-induced disruption of normal functioning of the body and psychogenic depressive changes.
Conclusions
During our study the stages of morphogenesis HIV-associated ulcerative changes in the periodontal tissues and OMM were firstly showed. We found that epithelial desquamation disrupts in patients with periodontal diseases and HIV-infection, which is a testament of the protective role reduce of OMM. In addition there is a compensatory-adaptive hyperkeratotnic alteration of the epithelium, as a precursor to AIDS-indicator conditions and display of maximum stress reserve of the organism. These data are confirmed by the results of the of keratinization index calculation. Its value increases along with the progression of the underlying disease and periodontal diseases.
Thus, the possibility of cytological diagnosis of oral tissues of people with HIV status can objectify pathological and functional changes in OMM in general and gums in particular. It helps to improve the efficiency of early objective diagnosis of pathological processes progression in periodontal tissues.
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Матер/'ал надйшов доредакци 26.03.2013р.
