
Оригинальные исследОвания ORIGINAL RESEARCH Экспериментальная медицина Experimental medicine © Group of authors, 2018 UDC 616.313.014.01 DOI – https://doi.org/10.14300/mnnc.2018.13095 ISSN – 2073-8137 THE HISTOCHEMICAL FEATURES OF THE ENDOTHELIUM OF THE TOOTH-ALVEOLAR COMPLEX IN INFLAMMATION AND OSTEOPOROSIS Sirak S. V. 1, Shchetinin E. V. 1, Bykova N. I. 2, Petrosyan G. G. 1, Tarabrina A. G. 1, Gayvoronskaya T. V. 2, Skorikova L. A. 2, Risovanniy S. I. 2 1 Stavropol State Medical University, Russian Federation 2 Kuban State Medical University, Krasnodar, Russian Federation ГИСТоХИмИЧЕСКИЕ оСоБЕННоСТИ ЭНдоТЕЛИя СоСУдов ЗУБоаЛЬвЕоЛярНоГо КомПЛЕКСа ПрИ воСПаЛЕНИИ И оСТЕоПороЗЕ С. в. Сирак 1, Е. в. Щетинин 1, Н. И. Быкова 2, Г. Г. Петросян 1, а. Г. Тарабрина 1, Т. в. Гайворонская 2, Л. а. Скорикова 2, С. И. рисованный 2 1 Ставропольский государственный медицинский университет, российская Федерация 2 Кубанский государственный медицинский университет, Краснодар, российская Федерация The results of immunohistochemical studies of the pulpal state and vascular endothelium of the dental alveolar complex are presented in the experimental modeling of periodontitis and osteoporosis in sheep. The endothelial structural param- eters of PD in animals in the gums of the upper and lower jaws differ and affect the length, thickness of cells, number of mitochondria, Golgi complex, reticulum of the endothelium, relationship between the nucleus and cytoplasm, and amount of heterochromatin in the nucleus. In periodontitis, pathological changes are noted in the endothelial circle, as well as in peri- cytes; this is manifested by a complex vascular-tissue reaction of the microcirculatory bed of periodontal tissues with a vio- lation of the rheological properties of the blood, combined with the formation of exudate and an inflammatory cell infiltrate. In the early period after the formation of the model of experimental osteoporosis, slight thickening and straightening were observed in the Korff fibers; at 180 days, the APhF exhibited a more linear structure, occasionally forming a finely mottled network that resembled a mosquito net. At 1 year of observation, there was a lack of Korff fibers in the primary substance of the pulp, and significant thickening of the argyrophilic membranes in the vascular endothelium; these are signs of impairment in the metabolism of the tooth pulp. Keywords: osteoporosis, periodontitis, experiment, endothelium Представлены результаты иммуногистохимического исследования пульпы и эндотелия сосудов зубоальвеолярно- го комплекса при экспериментальном моделировании пародонтита (ПД) и остеопороза (ОП) у овец. Эндотелиальные структурные показатели при ПД у животных в сосудах десен верхней и нижней челюсти различаются и затрагивают длину, толщину клеток, количество митохондрий, комплекс Гольджи, ретикулум эндотелия, взаимосвязь ядра и цито- плазмы и количество гетерохроматина в ядре. При ПД патологические изменения отмечены не только в эндотелиаль- ном кругу, но и в перицитах, что проявляется комплексной сосудисто-тканевой реакцией микроциркуляторного русла тканей пародонта с нарушением реологических свойств крови, образованием экссудата и воспалительного клеточно- го инфильтрата. В ранние сроки после формирования модели экспериментального ОП наблюдались незначительное утолщение и выпрямление волокон Корфа, а к 180 суткам эксперимента аргирофильные волокна становились более выпрямленными, иногда образовывая мелкопетлистую сеть, похожую на москитную сетку. В срок наблюдения 1 год в части препаратов наблюдалось отсутствие волокон Корфа в основном веществе пульпы, значительное утолщение аргирофильных мембран в эндотелии сосудов как признак ухудшения обмена веществ в пульпе зуба. Ключевые слова: остеопороз, пародонтит, эксперимент, эндотелий For citation: Sirak S. V., Shchetinin E. V., Bykova N. I., Petrosyan G. G., Tarabrina A. G., Gayvoronskaya T. V., Sko- rikova L. A., Risovanniy S. I. THE HYSTOCHEMICAL FEATURES OF THE ENDOTHELIUM OF THE TOOTH-ALVEOLAR COMPLEX IN INFLAMMATION AND OSTEOPOROSIS. Medical News of North Caucasus. 2018;13(3):520-525. DOI – https://doi.org/10.14300/mnnc.2018.13095 Для цитирования: Сирак С. В., щетинин Е. В., Быкова Н. И., Петросян Г. Г., Тарабрина А. Г., Гайворонская Т. В., Скорикова Л. А., Рисованный С. И. ГИСТОХИМИЧЕСКИЕ ОСОБЕННОСТИ ЭНДОТЕЛИЯ СОСУДОВ ЗУБОАЛЬВЕО- ЛЯРНОГО КОМПЛЕКСА ПРИ ВОСПАЛЕНИИ И ОСТЕОПОРОЗЕ. Медицинский вестник Северного Кавказа. 2018;13(3): 520-525. DOI – https://doi.org/10.14300/mnnc.2018.13095 520 МЕДИЦИНСКИЙ ВЕСТНИК СЕВЕРНОГО КАВКАЗА Medical news of north caucasus 2018. Т. 13. № 3 2018. Vоl. 13. Iss. 3 APhF – Argyrophilic Fibers IHCS – Immunohistochemical Study BM – Basal Membrane OP – Osteoporosis DAC – Dental Alveolar Complex PD – Periodontitis DAS – Dental Alveolar Segment nflammatory periodontal disease (PD) is an urgent of alcohol and xylene, after which they were immersed problem in dentistry. Due to its social importance, in Histomix histological medium (BioVitrum, Russia), IPD occupies a special role in medicine; according using a closed-type histological processor Tissue-Tek to recent data, up to 90 % of the population exhibit VIP™ 5 Jr and the paraffin filling station Tissue-Tek® PD of mild, moderate, and severe degrees [1, 2]. TEC™ 5 (Sakura, Japan). Histological sections of 3– The age spectrum of PD disease primarily covers a 5-μm thick were made from the resulting blocks by using period of 25–35 years; with increased age, severe a microtome and a table for the preparation of histological forms of PD are increasingly observed. Although the sections (Bio-Optica, Italy); the sections were stained nosology of PD has been involved a large number of with dyes (Bio-Optica and BioVitrum) on an automatic studies, there is no consensus regarding its etiology Prisma™ multi-stainer (Sakura, Japan). The painted and pathogenesis. In recent years, scientists have sections were enclosed in the mounting environment of speculated that a cause of PD involves circulatory Biomount (BioVitrum). Staining of sections for review disorders; in particular, dystrophic processes purposes was performed with hematoxylin and eosin; develop as a result of atherosclerotic changes in collagen fibers were stained by the Akimchenko and blood vessels [3, 4]. Notably, pathomorphological Masson methods. To identify antigens in tissue cells, studies revealed significant changes in the walls IHC was performed with antibodies: monoclonal mouse of the periodontal arteries (perivascular sclerosis, antibodies to vimentin (V9), α-smooth muscle actin-α- thickening of the walls of blood vessels, narrowing of SMA (1A4), CD34 (QBEnd/10), epithelial membrane the lumen, and/or hyalinosis). antigen (Anti-EMA, E29) (Spring Bio Science, USA). IHC In the scientific literature, a pathological role of reactions were performed with paraffin sections by using hypoxia in PD has been highlighted. Multiple studies have the highly sensitive Reveal biotin-free polyvalent DAB revealed a reduction in oxidation-reduction potential in imaging system (Spring Bio Science). The intensity of periodontal pathology, as evidenced by the accumulation IHC in each sample was monitored under a microscope. of weakly oxidized products in tissues; as a result of For the second stage of the study, 10 sheep were this change, the barrier function of periodontal tissues divided into two groups: basic (six animals) and control decreases and pathological phenomena characteristic (four animals). In the main group, an experimental model of PD (e.g., gingivitis, bone resorption, and pathological of OP was induced under general anesthesia (Zoletil 50); periodontal pocket formation) occur [5]. Of particular ovariectomy was performed, followed by intramuscular interest is the study of the histochemical features of injection of dexamethasone for 3 months [9]. Tissue periodontal tissues in a chronically progressing systemic collection was performed under general anesthesia in the metabolic disease of the skeleton: osteoporosis (OP), area of the lower incisors (hooks), by isolating DAS under which is characterized by a reduction in bone density, general anesthesia after 90 days, 180 days, and 1 year a violation of microarchitectonics, and an increase in after the start of the experiment. In the control group, the fragility. corresponding dental alveolar complex (DAC) in the intact The aim of this study was to perform a histochemical animals was examined. The isolated blocks were fixed for evaluation of the structure of the vascular endothelium in 8–10 days in a 12 % solution of neutral formalin, decalcified periodontal tissues in inflammation and OP. in a mixture of equal volumes of 10 % formic and 8 % Material and Methods. Experimental research was hydrochloric acid, washed under running water, immersed performed in two stages; experimental animals were in alcohol solutions of increasing concentration, and poured mature sheep of the North Caucasian breed (18 animals, into celloidin. Sections 3–5-μm-thick were stained with including 10 females and 8 males). During the study, two hematoxylin and eosin, then impregnated with silver by the experimental models were created in animals: PD and OP. Fout method. When APhF was detected in the decalcified During the first stage of the experiment, four animals blocks, slices were prepared to assess the state of the with unchanged periodontium (control group) and four cellular elements of the tooth pulp by impregnating with with obvious signs of disease (the main group) were silver nitrate by the Bilshovsky-Gross and
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