Кишинев.
Chisinau.
Катеренюк Илья Мафтеевич.
бул. Штефан чел Маре, 165, г. Кишинев, MD-2004, Республика Молдова.
Ilia Catereniuc.
bd. Ştefan cel Mare şi Sfânt, 165, Chisinau, MD-2004, Republic of Moldova.
Тимишоара.
Timisoara.
Тимишоара.
Timisoara.
Целью данного исследования было изучение vasa venarum большой подкожной вены ноги, эндо- телиальной пролиферации и оценка изменения плотности микрососудов при атеросклерозе и ассоциированном сахарном диабете.
Материал и методы. Изучено 9 фрагментов подкожных вен человека, взятых у пациентов, кото- рым ампутировали нижние конечности. Причинами ампутации были: травма (1 случай) и атеросклероз, осложненный окклюзией или тромбозом артерий нижних конечностей (8 случаев). Среди случаев атеросклероза 2 были ассоциированы с сахарным диабетом. Фиксированные формалином ткани заливали в парафин и готовили серийные срезы толщиной 3 мкм. Полученные срезы окрашивали методом двойного иммуноокрашивания с использованием моноклональных антител анти-CD34 и анти-Ki67. Плотность микрососудов определяли путем подсчета сосудов в трех «hot spots», средние значения которых были разделены на поверхности поля высокой мощности разрешения (0.0625 мм2). Пролиферативный индекс эндотелиальных клеток рассчитывали в процентном отношении ко всем окрашенным Ki67 ядрам эндотелиальных клеток, которые также ко-экспрессировали позитивное цитоплазматическое окрашивание в CD34- позитивных клетках.
Результаты. В исследованных фрагментах больших подкожных вен человека, vasa venarum были обнаружены в наружной и средней оболочках. При травматическом повреждении нижней конечности адвентициальные vasa venarum были в 4.5 раза многочисленнее (299 сосудов/мм2), чем в средней оболочке больших подкожных вен (64 сосуда/мм2). В случае сопутствующих метаболических нарушений было обнаружено увеличение количества vasa venarum в средней оболочке подкожной вены, 247±109 сосудов/мм2 и 253±51 сосуд/мм2 соответственно. Однако, при атеросклерозе (151±23 сосуда/мм2) и ассоциированном сахарном диабете (125±3 сосуда/мм2) плотность адвентициальных микрососудов была ниже чем в средней оболочке. Пролиферативный индекс эндотелиальных клеток в подкожной вене у пациента с травматическими повреждениями составлял 20%–25%, в то время как у пациентов с нарушениями обмена веществ был выше в диапазоне от 25% до 30%. В адвентиции и медии наблюдались как ангиогенез путем почкования, «sprouting», так инвагинационный «non-sprouting» ангиогенез.
Выводы. В нормальных условиях микроциркуляторные сосуды преобладают в адвентиции, а при патологических состояниях (атеросклерозе, сахарном диабете) микрососуды средней оболочки могут численно превосходить микрососуды адвентиции. Vasa venarum средней оболочки чаще располагаются в ее наружном слое. Пролиферативный индекс эндотелия Ki67 выше (25–30%) в случае атеросклероза и ассоциированного сахарного диабета, чем в нормальных условиях (20–25%). Образование и рост новых микрососудов, neovasa vasorum, происходит как путем «sprouting», так и «non-sprouting» ангиогенеза.
The aim of the study was to investigate vasa venarum of great saphenous vein and their endothelial proliferation and to evaluate the modification of the microvessel density in case of atherosclerosis and associated diabetes mellitus.
Material and methods. The present study included nine venous fragments of human saphenous veins collected from the patients whose lower limbs were amputated. The causes of the limb ablation were: trauma (1 case) and atherosclerosis complicated with occlusion or thrombosis of the lower limbs arteries (8 cases). There were two cases presenting associated diabetes mellitus among the eight cases of atherosclerosis. Formalin-fixed tissues were paraffin embedded and cut into 3μm transverse sections. The slides were stained by a double immunostaining technique using the monoclonal antibodies anti-CD34 and anti-Ki67. The microvessel density was determined by counting the vessels of three “hot spots” and represented by the average values of three “hot spot” areas divided on the surface of high power field (0.0625 mm2). The endothelial cell proliferation index was calculated as the percentage of all nuclei of Ki67-stained endothelial cells that also co-expressed positive cytoplasmic staining in CD34-positive cells.
Results. Vasa venarum of the examined segments of human great saphenous veins were found both in the adventitia and the tunica media. In case of the traumatic injury of the lower limb the adventitial vasa venarum were 4.5 times more numerous (299 vessels/mm2) than those of great saphenous vein media (64 vessels/mm2). In case of associated metabolic disorders we found increased number of vasa venarum of saphenous vein media, 247±109 vessels/mm2 and 253±51 vessels/mm2, respectively. However, the adventitial microvessel density was lower in case of atherosclerosis (151±23 vessels/mm2) and associated diabetes mellitus (125±3 vessels/mm2). The endothelial cell proliferation index was between 20% and 25% in saphenous vein from the patient with the traumatic injury. Moreover, the proliferation index was higher, ranged between 25% and 30%, in saphenous vein from the patients with metabolic disorders. In the adventitia and the media sprouting and non-sprouting angiogenesis were observed.
Conclusions. Under normal conditions, the microcirculatory vessels predominate in the adventitia, but under pathological conditions (atherosclerosis, diabetes mellitus) the microvessels of the media can exceed numerically the microvessels of the adventitia. The vasa venarum of the tunica media can be located in its outer layer. The Ki67-endothelial proliferation index can be higher (25–30%) in case of the atherosclerosis and associated diabetes mellitus than that in normal conditions (20–25%). The formation of the neovasa vasorum occur through both sprouting and non-sprouting angiogenesis.
The aim of the study was to investigate vasa venarum of great saphenous vein and their endothelial proliferation and to evaluate the modification of the microvessel density in case of atherosclerosis and associated diabetes mellitus.
Material and methods. The present study included nine venous fragments of human saphenous veins collected from the patients whose lower limbs were amputated. The causes of the limb ablation were: trauma (1 case) and atherosclerosis complicated with occlusion or thrombosis of the lower limbs arteries (8 cases). There were two cases presenting associated diabetes mellitus among the eight cases of atherosclerosis. Formalin-fixed tissues were paraffin embedded and cut into 3μm transverse sections. The slides were stained by a double immunostaining technique using the monoclonal antibodies anti-CD34 and anti-Ki67. The microvessel density was determined by counting the vessels of three “hot spots” and represented by the average values of three “hot spot” areas divided on the surface of high power field (0.0625 mm2). The endothelial cell proliferation index was calculated as the percentage of all nuclei of Ki67-stained endothelial cells that also co-expressed positive cytoplasmic staining in CD34-positive cells.
Results. Vasa venarum of the examined segments of human great saphenous veins were found both in the adventitia and the tunica media. In case of the traumatic injury of the lower limb the adventitial vasa venarum were 4.5 times more numerous (299 vessels/mm2) than those of great saphenous vein media (64 vessels/mm2). In case of associated metabolic disorders we found increased number of vasa venarum of saphenous vein media, 247±109 vessels/mm2 and 253±51 vessels/mm2, respectively. However, the adventitial microvessel density was lower in case of atherosclerosis (151±23 vessels/mm2) and associated diabetes mellitus (125±3 vessels/mm2). The
endothelial cell proliferation index was between 20% and 25% in saphenous vein from the patient with the traumatic injury. Moreover, the proliferation index was higher, ranged between 25% and 30%, in saphenous vein from the patients with metabolic disorders. In the adventitia and the media sprouting and non-sprouting angiogenesis were observed.
Conclusions. Under normal conditions, the microcirculatory vessels predominate in the adventitia, but under pathological conditions (atherosclerosis, diabetes mellitus) the microvessels of the media can exceed numerically the microvessels of the adventitia. The vasa venarum of the tunica media can be located in its outer layer. The Ki67-endothelial proliferation index can be higher (25–30%) in case of the atherosclerosis and associated diabetes mellitus than that in normal conditions (20–25%). The formation of the neovasa vasorum occur through both sprouting and non-sprouting angiogenesis.
The authors declare that there are no conflicts of interest present.