Cytomorphological assessment of buccal epithelium of the cheeks of children of some industrial cities of the Karaganda region

The article discusses changes in the buccal epithelium of the cheeks (BECh) based on the results of a survey of the child population of three industrial cities of the Karaganda region (Temirtau, Balkhash, Abay), methods of cytological analysis were used. Cytochemical methods were used for the early detection of functional changes in the nonspecific protection of the body from environmental factors. When conducting a research on the cytological status of buccal epithelium cells of children living in the studied areas, it was revealed that an increase in the number of cells with small-droplet inclusions in the cytoplasm of children of Temirtau and Balkhash was found. When comparing the results of investigation of child population (Temirtau, Balkhash, Abay), a decrease in the number of cells with small-droplet inclusions in the cytoplasm by 4 times and an increase in the number of cells with large-droplet inclusions in the cytoplasm for 78 %, with signs of karyorexis for 74 % was noted. The number of nuclear-free cells was reduced for 34 %, degenerated neutrophil leukocytes by 3.9 times. There is an increase in the amount of amitosis for 50 % and a decrease in the number of normal buccal cells for 71 %. The appearance of small and large inclusions in the cytoplasm of cells proves the effect of adverse environmental factors on the body of children living in industrial regions.

Introduction

Indicators of the functional state of some body systems should be considered more informative than the incidence, when assessing the health status of the population. Thus, minimal changes in the quality indicators of buccal epithelial cells of the cheeks of children were noted at a pollution level approaching 3 MPCs, and at the same time they did not have significant changes in the incidence of allergic diseases and changes in immune reactivity [1-3]. Children, living in areas with unfavorable hygienic conditions, have lowered functional reserves of the body, and changes in their respiratory parameters prevail. Under the influence of adverse environmental factors (water, food, atmospheric air), subnormal changes in the body of children were recorded [4, 5].

Numerous studies and the results of cytomorphological indicators of the BECh of the child population [6-10] have shown prospects for scientific research in this area. The cytological state of the BECh reflects the state of the body, changing in dependence of the pollution of the production environment. Studies have shown that the epithelium of the mucous membranes of different degrees of differentiation, vary from different adverse effects and can be considered as a target for these effects [11, 12]. Non-invasive multi-organ micronucleus test on human exfoliating cells, including analysis of buccal epithelial cells, allows to verify the data, obtained from biological material, as well as to expand the amount of information about the existing adverse factors. In this case, the cheek mucosa is under the predominant influence of factors acting under inhalation effect [3, 13, 14].

Methodology

Based on this, a research work of children of primary classes of secondary schools in Temirtau, Balkhash and Abay of Karaganda region was conducted. In total 240 children at the age of 6-8 years were examined. Methods of cytological analysis of BECh with the help of MS-200 microscope (Austria, 2004) were used, Kolmogorov-Smirnov criterion was used to evaluate normality of data distribution. Arithmetic mean, variance, error, and 95 % confidence interval were calculated for quantitative variables with normal distribution. Differences between the groups were revealed with the help of methods of parametric statistics. Comparison and assessment of relative risks was performed using the value of χ2. Non-parametric criteria based on the one-factorial dispersive analysis based on Wilcoxon rank labels, the median test, and Spearman's rank correlation coefficient were used for the calculation [15-17]. The results of the deviation were compared with the physiological limits of fluctuation. When calculating the BECh values, 200 cells were analyzed, 300 cells were counted if many different anomalies were recorded. During the analysis, 75-98 normal epithelial cells were taken as «normal». The number from 55 to 74 normal epithelial cells indicates an initial decrease in body resistance. A quantity from 15 to 54 and below indicates a decrease in body resistance. Incomplete phagocytosis (microbial digestion, cell residue) — reduced immunity, karyorrhexis indicates cell necrosis [9, 14].

Results and their discussion

When conducting a research on the cytological status of buccal epithelium cells of children living in the studied areas, it was found that in Temirtau and Balkhash children have the increased number of cells with droplets in the cytoplasm which is 0.47 and 1.90, accordingly. At the same time, the indicators of epithelial cells with large-droplet inclusions of children living in Temirtau are 2.1. This figure is lower for children living in the city of Balkhash and is 1.2.

It was found that the cytomorphological indicator of a cell with signs of karyorrhexis of children living in Temirtau is 1.7 times more than of children living in Balkhash, the same indicator is 1.7 times higher in the Balkhash group of children than in Abay. The number of degenerated neutrophilic leukocytes (DNL) of the Balkhash group of children is increased by 2.5 times, and of children of the Temirtau group is decreased relative to the group of children from the city of Abay. The number of normal buccal cells of the cheeks in both groups was reduced: the Balkhash group by 1.4 and the Temirtau group by 2.4 times. The percentage of microflora contamination (streptococci, staphylococcus) in the 2nd and 3rd groups was increased by 26 % and 2.8 times (Fig. 1–3).

The functional status of buccal cells depends on the degree of their maturity, as in other epithelial cells. As part of the multilayer flat epithelium, buccal epithelial cells are at different stages of morphological and functional differentiation-from poorly differentiated precursors in the basal layer (they provide regeneration of the epithelium) to highly specialized cells, which while being differentiated are shifted to the surface layers, undergoing desquamation, some of them have signs of keratinization.

Differentiated, proliferative processes and functional parameters of mature cells are regulated by factors of local and central origin. It is not surprising that the state of buccal epithelial cells reflects the nature of destabilizing processes at the local and systemic levels [5, 12, 14, 18]. Changes in the differentiation of the epithelium, recorded morphologically (cell size, nature of nuclei and granules, signs of cytolysis) and electrokinetically (electrical mobility of nuclei), are proposed to be taken into account in screening assessment of health, stressful effects of harmful environmental factors, somatic pathology, biological age of a person.

The functional characteristics of epithelial cells also include such an important indicator as the ability for adhesive interactions with microorganisms. On this depends not only the nature of the microbial colonization of the epithelium and the state of local immunity, but also the homeostasis of the entire cellular community associated with the mucous membranes. Streptococci dominate in the obligate microflora of buccal cells. Their number is maximum in children under 10 years of age and is considered as one of the indicators of nonspecific resistance of the oral mucosa and «general health». The presence of less typical and atypical microorganisms for a given biotope reflects the weakening of colonization resistance, signaling destabilization processes, focused on the level of mucous membranes [18, 19].

A study of the buccal epithelium cells of the cheeks of the examined child population recorded a sixfold increase in the number of phagocytosed apoptotic (residual) bodies in all urban children. The number of cells with vacuolar dystrophy was above the norm in all examined urban children of Abay, Balkhash, Temirtau (Fig. 4).

The results showed that children living in the cities of Abay, Balkhash, Temirtau, despite the different place of residence, have catarrhal and atrophic rhinitis, because the buccal mucosa of the cheeks are the most

important biological barriers that prevent adverse and anthropogenic factors from entering the body. On the buccal epithelium of the cheeks the reparative process disorders were found, which affects their ability to interact adhesively with microorganisms constantly residing in the oral cavity, which leads to their accumulation. Inability to neutralize microorganisms indicates a disorder of local immunity [3, 6, 8]. Moreover, the buccal mucosa reflects the immune system of the entire digestive tract. Most of the nutrients enter the body through the gastrointestinal tract, the adequate functioning of which is ensured by normal microbiocenosis. [14, 18]. In the event of a change in microbiological associations, a negative change occurs in the processes of absorption, intestinal dysbiosis. Due to the toxic-dysmetabolic effects on the body, the formation of pathological processes is possible, which requires further in-depth study of the damaging effects of chemical factors. [12, 14, 19]. Malabsorption of essential trace elements occurs as a result of dismetabolic processes in the intestine, even with sufficient use of trace elements with food [12, 14]. Pediatricians note that prescribing iodine-containing drugs does not correct the iodine deficiency condition. This phenomenon is possible for any deficiency of trace elements (zinc, selenium). The revealed changes in the cytomorphological indices of the buccal epithelium make it possible to recommend non-invasive analyzes of the nasal epithelium, along with the buccal, to assess the effects of adverse environmental factors, especially to detect the inhalation effect of air pollutants.

References

1. Sviatov, S.S., & Tamenova, S.S. (2014). Ekolohicheskie problemy v stranakh Tsentralnoi Azii [Environmental problems in the countries of Central Asia]. Moscow: Izdatelstvo Moskovskoho universiteta [in Russian].
2. Bazeliuk, L.T., Hazalieva, M.A., & Saparhalieva, S.K. (2008). Tsitomorfolohicheskaia i metabolicheskaia otsenka bukkalnoho epiteliia shchek u rabochikh berillievoho proizvodstva h. Ust-Kamenohorska [cytomorphological and metabolic assessment of buccal epithelium of cheeks of workers of beryllium production in Ust-Kamenogorsk]. Zdorove i bolezn — Health and disease, 1, 35–39 [in Russian].
3. Namazbaeva, Z.I., Duzbaeva, N.M., & Bazeliuk, L.T. (2008). Tsitomorfolohicheskie osobennosti rinotsitohrammy i bukkalnoho epiteliia u shkolnikov srednei shkoly «Daryn» horoda Karahandy [cytomorphological features of rhinocytogram and buccal epithelium schoolchildren of the secondary school «Daryn» in Karaganda]. Proceedings from Efektivni nastroje modernich ved – 2008 [Effective Instruments of Modern Sciences-2008]: IV mezinarodni vedecko-prakticka konference — IV International Scientific and Practical Conference (03–15 kvetna 2008 roku) [03–15 May 2008 year] (pp. 51–54). Praha: Education and Science [in Russian].
4. Bazeliuk, L.T., & Eshmahambetova, A.B. (2012). Tsitomorfolohicheskaia otsenka rinotsitohrammy i bukkalnoho epiteliia shchek u detei podverhaiushchikhsia khimicheskoi nahruzke v usloviiakh promyshlennoho horoda Temirtau [cytomorphological assessment of the rhinocytogram and buccal epithelium of cheeks of children undergone through chemical influence in conditions of industrial city Temirtau]. Proceedings from Theoretical and practical aspects of modern medicine: Mezhdunarodnaia nauchno- prakticheskaia konferentsiia — International scientific and practical conference (pp. 83–86). Novosibirsk [in Russian].
5. Namazbaeva, Z.I., Sabirov, Zh.B., & Oblezina, A.V. (2012). Tsitoheneticheskii status podrostkov, prozhivaiushchikh na territorii promyshlennoho horoda [cytogenetic status of adolescents living on the territory of an industrial city]. Proceedings from Theoretical and practical aspects of modern medicine: Mezhdunarodnaia nauchno-prakticheskaia konferentsiia — International scientific and practical conference (pp. 78–83). Novosibirsk [in Russian].
6. Volkova, A.T., & Viktorova, T.V. (2011). Sravnitelnyi analiz tsitoheneticheskoi nestabilnosti kletok bukkalnoho epiteliia u horodskikh i selskikh zhitelei Respubliki Bashkortostan [comparative analysis of cytogenetic instability of buccal epithelium cells in urban and rural residents of the Republic of Bashkortostan]. Hihiena i sanitariia — Hygiene and sanitation, 5, 40–42 [in Russian].
7. Beliaeva, N.N., Shamarin, A.A., Petrova, I.V., & Malysheva, A.H. (2001). Sviaz izmenenii slizistykh obolochek nosa i rta s immunnym statusom pri vozdeistvii faktorov okruzhaiushchei sredy [The connection of changes in the mucous membranes of the nose and mouth with the immune status under the influence of environmental factors]. Hihiena i sanitariia — Hygiene and sanitation, 5, 62–64 [in Russian].
8. Aleksandrova, V.P. (2010). Analiz sostoianiia zdorovia i tsitolohicheskoho statusa slizistykh obolochek nosa i shcheki pri otsenke korrektsii vodopotrebleniia u detei [Analysis of the health status and cytological status of the mucous membranes of the nose and cheek in assessing the correction of water consumption in children]. Extended abstract of candidate's thesis. Moscow [in Russian].
9. Beliaeva, N.N., Sycheva, L.T., Zhurkov, V.S., & Shamarin, A.A. et al. (2005). Otsenka tsitolohicheskoho i tsitoheneticheskoho statusa slizistykh obolochek nosa i rta u cheloveka [Assessment of the cytological and cytogenetic status of the mucous membranes of the nose and mouth in humans]. Moscow [in Russian].
10. Duzbaeva, N.M., Bazeliuk, L.T., & Zeinidenov, A.K. (2011). Sostoianie metabolicheskoho statusa slizistoi obolochki nosa, bukkalnoho epiteliia shchek i perifericheskoi krovi u detei, prozhivaiushchikh v raionakh tekhnohennoho vliianiia [condition of the metabolic status of the nasal mucosa, buccal epithelium of cheeks and peripheral blood of children living in areas of anthropogenic influence]. Vestnik Karahandinskoho universiteta. Seriia Biolohiia. Meditsina. Heohrafiia — Bulletin of the Karaganda University. Series Biology. Medicine. Geography, 1, 13–15 [in Russian].
11. Sidorova, I.E., Revazova, Yu.A., & Safonov, V.V. (2004). Izuchenie heneticheskoho polimorfizma i tsitoheneticheskikh narushenii u lits, imevshikh kontakt s toksichnymi khimicheskimi soedineniiami [The study of genetic polymorphism and cytogenetic disorders in persons who had contact with toxic chemical compounds]. Donozolohicheskaia diahnostika — Donosological diagnostics, 6, 59–62 [in Russian].
12. Abadzhi, M.A., Molodtsov, S.A., & Ashkinazi, V.I. et al. (2002). Mikroflora bukkalnoho epiteliia u detei, chasto boleiushchikh respiratornymi infektsiiami [Microflora of buccal epithelium in children who are often ill with respiratory infections]. Rossiiskii pediatricheskii zhurnal — Russian Pediatric Journal, 1, 56–57 [in Russian].
13. Namazbaeva, Z.I., Bazeliuk, L.T., Mukasheva, M.A., & Aitkulov, A.M. (2001). Informativnost biokhimicheskikh i tsitokhimicheskikh markerov u laboratornykh zhivotnykh pri naturnykh issledovaniiakh [Informativeness of biochemical and cytochemical markers in laboratory animals in field studies]. Hihiena i sanitariia — Hygiene and sanitation, 1, 20–22 [in Russian].
14. Abadzhi, M.A., Makhrova, T.V., & Maianskaia I.V. et al. (2003). Bukkalnye epiteliotsity kak instrument kliniko- laboratornykh issledovanii [Buccal epithelial cells as a tool for clinical laboratory studies]. Nizhehorodskii meditsinskii zhurnal — Nizhny Novgorod Medical Journal, 3–4, 105–110 [in Russian].
15. Gildenskiold, R.S., Yastrebov, G.G., & Vinokur, I.L. et al. (1996). Unifitsirovannye metody sbora dannykh, analiza i otsenki zabolevaemosti naseleniia s uchetom kompleksnoho deistviia faktorov okruzhaiushchei sredy [Unified methods of data collection, analysis and assessment of the incidence of the population, taking into account the complex effect of environmental factors]. Moscow [in Russian].
16. Gusarov, V.M. (2003). Statistika [Statistics]. Moscow: YuNITI-DANA [in Russian].
17. Nazarov, M.G. (Eds.). (2009). Statistika [Statistics]. Retrieved from https://static.my-shop.ru/product/pdf/208/2074707.pdf
18. Namazbaeva, Z.I., Aitkulov, A.M., Sabirov, Zh.B., & Ageev, D.V. (2014). Risk zdoroviu naseleniia pri vozdeistvii melkodispersnoi pyli atmosfernoho vozdukha slozhnoho khimicheskoho sostava [The risk to the health of the population when exposed to fine dust of atmospheric air of a complex chemical composition]. Proceedings from Zoology at the turn of the century: Mezhdunarodnaia nauchno-prakticheskaia konferentsiia — International Scientific and Practical Conference (pp. 15). Karaganda [in Russian].
19. Namazbaeva, Z.I., & Bazeliuk, L.T. (2014). Izmeneniia funktsionalnoho sostoianiia kletochnykh struktur u krys pri vozdeistvii vzveshennykh pylevykh chastits [changes in the functional state of cellular structures in rats under the influence of suspended dust particles]. Proceedings from Innovations in Science: Mezhdunarodnaia konferentsiia — International Conference (pp. 91–103) [in Russian].