Histological and immunohistochemical evaluation of mandibular bone tissue regeneration

The purpose of the study was to perform an immunohistochemical and histological evaluation of samples taken from different bone regeneration procedures in atrophic human mandible. 30 patients (15 men and 15 women, age range of 35-60 years), non-smokers, with good general and oral health were recruited in this study and divided into three groups. The first group included patients who were treated with blood Concentration Growth Factors (bCGF), the second group included patients who were treated with a mixture of bCGF and autologous bone, while the third group of patients was treated with bCGF and tricalcium phosphate/hydroxyapatite (TCP-HA). Six months after the regenerative procedures, all patients undergone implant surgery, and a bone biopsy was carried out in the site of implant insertion. Each sample was histologically and immunohistochemically examined. Histological evaluation showed a complete bone formation for group II, partial ossification for group I, and moderate ossification for group III. Immunohistochemical analysis demonstrated a statistically significant difference between the three groups, and the best clinical result was obtained with a mixture of bCGF and autologous bone

Корреляция плотности макрофагов с тяжестью неоплазии шейки матки

Despite all recent efforts, cancer of the uterine cervix still remains one of the most frequent malignancies among women. Lymphatic vessels represent the primary route of tumor cells dissemination in cervical cancer. It has been demonstrated that cervical neoplasia actively participates in the recruitment of new blood and lymphatic vessels. Macrophages are extremely versatile cells which have a significant contribution to tumor progression. The aim: 1) To establish the correlation between tumor-associated macrophages (TAM) and the grade of the uterine cervix neoplasia; 2) To evaluate the distribution of TAM within both intratumoral and peritumoral areas. Material and Methods: Ninety-six cases were studied. The specimens were fixed in buffered formalin and paraffin embedded. Step sections, 5μm thick, were performed for each case. Initial sections were stained with haematoxylin-eosin, for the pathological diagnosis and grading of the tumor. Lesions were classified as follows: squamous cell metaplasia (n = 12), CIN I (n = 8), CIN II (n = 6), CIN III (n = 24), microinvasive carcinoma (n = 16) and invasive squamous cell carcinoma (n = 26). Additional sections for each case were stained for CD68 antibody, in order to highlight the macrophages. Quantification of macrophage population has been made based on hot-spot technique. The arithmetic media of 3 (× 200) fields represented the final result. Results: We found a statistical correlation between peritumoral macrophages (PTM) and intratumoral macrophages in all stages of cervical neoplasia, macrophage density and tumor stage (p = 0.01). In 16 cases we found vascular invasion. Almost in all these cases (87.5%) intravascular tumor emboli were embedded with CD68 cells. Conclusions: based on these findings, we consider that macrophages are key regulators of cervical cancer progression. TAM targeted management could be an essential therapeutic strategy, not only in order to suppress the progression of cervical neoplasia, but also to inhibit macrophage-mediated vascular invasion.Рак шейки матки остается одной из самых часто встречающихся злокачественных заболеваний женского населения. Лимфатические сосуды являются первичным путем метастазирования при данном заболевании. Было доказано, что клетки цервикальной неоплазии активно участвуют в образовании новых лимфатических сосудов. Макрофаги – многофункциональные клетки, оказывающие большое влияние на прогрессирование опухоли. Цель: 1). Выявление корреляции между макрофагами и стадией прогрессии неоплазии шейки матки; 2). Определение особенностей распределения макрофагов внутри опухолевой массы и вокруг нее. Материал и методы. Было изучено 96 случаев. Материал фиксировали в формалине с последующим заключением в парафин. Для каждого случая производили срезы, толщиной в 5 мкм. Изначально, срезы окрашивали гематоксилин-эозином для определения гистопатологического диагноза. Были получены следующие группы поражений: плоскоклеточная метаплазия (n = 12), CIN I (n = 8), CIN II (n = 6), CIN III (n = 24), микрокарцинома (n = 16), инвазивный рак (n = 26). Для выявления макрофагов, произвoдили иммуногистохимическое исследование с использованием маркера CD68. Подсчет популяции макрофагов производили по методике hot-spot. Результаты. Мы получили статистически значимую корреляцию между внутритуморальными и перитуморальными макрофагами во всех стадиях прогрессии неоплазии шейки матки, между плотностью макрофагов и стадией опухоли (p = 0,01). В 16 случаях выявили сосудистые эмболы. Почти во всех случаях (87,5%) внутрисосудистые эмболы содержали в себе CD68 клетки. Выводы: Основываясь на полученных результатах, мы считаем, что макрофаги вовлечены в прогрессию рака шейки матки

Роль рецептора сосудистого эндотелиального фактора роста (VEGFR-3) в процессе канцерогенеза шейки матки

In spite of the huge achievements made in the early detection of uterine cervix cancer, this disease remains one of the most widespread human malignancies throughout the world. Nowadays, there is a tendency of cervical cancer to affect the youth population. Early metastasizing is a critical point in evolution of this disease. Lymphatic vessels (LV) represents the primary route of cancer cells spreading towards the distant sites. That is why understanding of new lymphatics recruitment by the tumor, at molecular level, could be a potent support in solving of metastasizing. VEGFR-3 is a tyrosine-kinase receptor, specific for lymphatic endothelial cells, that being activated provides their proliferation, surviving and migration. It has been demonstrated that VEGF-R3 mediates invasion activity of tumor cells into surrounding stroma and vascular structures. The aim of this research was to study the importance of VEGFR-3 in cervical carcinogenesis. Material and methods: We studied biological material, taken by targeted biopsy and conization, from women with detectable cervical lesions. Detection VEGFR-3 was made using monoclonal antibody anti VEGF-R3 through Avidin-Biotin working system, LSAB technique. Lymphatic microvascular density was assessed by podoplanin labeling, using anti D2-40. A statistical analysis was made with SPSS 13.0. Results: We obtained VEGFR-3 expression in LV, blood vessels, tumor mass and stromal cells. The highest density of VEGFR-3 LV was in CIN III with gradually decreasing in invasive stages. In invasive carcinoma we obtained statistically significant correlation between tumor VEGFR-3 expression and vascular invasion. Conclusions: VEGFR-3 is not specific for lymphatic endothelium. It can not be used for alone lymphatic microvascular density assessing. It is actively involved in vascular invasion.Несмотря на огромный успех, достигнутый в выявление рака шейки матки (РШМ) на ранних стадиях развития, данное заболевание остается одной из самых часто встречающихся злокачественных новообразований во всем мире. На сегодняшний день наблюдается тенденция омоложения РШМ. Раннее метастазирование является ключевым моментом в эволюции заболевания. Лимфатические сосуды представляют собой первичный путь распространения опухолевых клеток. В связи с этим, понимание механизмов образования лимфатических сосудов на молекулярном уровне, может помочь решить эту проблему. VEGFR-3 является тирозин-киназным рецептором. Он находится на поверхности лимфатических эндотелиальных клеток, при активации которого происходит их пролиферация, миграция и выживание. Было доказано, что VEGFR-3 регулирует инвазию раковых клеток в строму и сосуды. Цель работы: изучить роль VEGFR-3 в канцерогенезе РШМ. Материал и методы: был изучен биологический материал, полученный посредством прицельных биопсий и конизаций у пациенток с макроскопически выявленным поражением шейки матки. Для определения VEGFR-3 была использована система Avidin-Biotin, техника LSAB , с применением первичного антитела анти VEGFR-3. Для определения лимфатических сосудов были использованы моноклональные антитела анти D2-40. Статистический анализ данных был произведен при помощи SPSS 13.0. Результаты: мы выявили экспрессию VEGFR-3 не только в лимфатических сосудах, а также в кровеносных сосудах, в опухолевой массе и клетках стромы. Наибольшая плотность лимфатических сосудов была определена на стадии CIN III (интраэпителиальная цервикальная неоплазия), с последующим прогрессивным уменьшением. На стадии инвазивного рака мы получили статистически значимую корреляцию между интенсивностью экспрессии VEGFR-3 опухолью и сосудистой инвазией, а также между интенсивностью экспрессии VEGFR-3 опухолью и внутрисосудистыми опухолевыми эмболами. Выводы: VEGFR-3 не является специфическим маркером для лимфатического эндотелия. Данный маркер не может использоваться самостоятельно для определения лимфатической микрососудистой плотности. VEGFR-3 активно вовлечен в процесс сосудистой инвазии

Expression of CK5 basal cytokeratin in primary breast carcinoma

Department of Histology, Cytology and Embryology, Laboratory of Morphology, Nicolae Testemitsanu State University of Medicine and Pharmacy, Chisinau, the Republic of MoldovaBackground: CK5 positive cells represent progenitors for glandular and myoepithelial lineages of mammary epithelium. During epithelial differentiation there is a gradual decrease of CK5 expression. In case of benign lesions the proliferating luminal cells show a high expression of CK5. Contrary, the majority of malignancies which are derived from differentiated glandular cells line do not reveal immunohistochemical staining with CK5 marker. The aim of this study was to compare the expression of basal cytokeratin CK5 vs hormone receptors, HER2, Ki67 and molecular subtype’s immunohistochemically defined in the primary breast carcinomas of NST type. Material and methods: We processed 108 invasive breast carcinomas of NST type. The specimens were formalin-fixed and paraffin-embedded as traditionally. Sections were immunostained (ER, PR, HER2, CK5 and Ki67) automatically with Leica Bond-Max autostainer. Results: Breast carcinoma of NST type was in majority of cases CK5 negative (94 cases/87%). The positive CK5 cases had a high grade of differentiation. CK5 negative tumors were usually hormone positive, but in 8 cases/6.5% a combined simultaneous CK5-ER (PR) positive expression was determined. From 22 HER2 positive cases, 16 were CK5 negative. CK5 value correlated statistically significant with all used markers, except grade of differentiation: a positive Pearson coefficient was determined in relation to HER2 and Ki67, and a negative one compared to hormone receptors and molecular subtype. Conclusions: We support CK5 potential value in molecular subtype’s differentiation. Breast carcinoma of NST type is usually CK5 negative and hormone positive. The presence of cases with simultaneous expression of CK5 and hormone receptors is an open field to debate the existence of other, transient molecular subtypes and we expect a further confirmation in larger study groups

Immunohistochemical characteristic of cellular and humoral immunity and preimmune factors in chronic tonsillitis in children

AbstractIn order to exclude errors in choosing the treatment strategy in case of chronic tonsillitis it needs to develop new contemporary diagnostic and prognostic criteria. [1, 2] The purpose of our research was to determine immunohistochemical peculiarities of the palatine tonsils during the evolution of different forms of chronic tonsillitis in children. Immunohistochemical study were subjected tissue samples taken from palatine tonsils from 21 children with compensated chronic tonsillitis and 20 children with decompensated form, the last being subdivided into two forms: without late complications and with rheumatic complications