Abstract

Objective - to study the morphological features of platelets of newborns by using electron microscopy. Materials and methods. Peripheral blood platelets of healthy newborns were studied by means of transmission electron microscopy (TEM) method. The sample included 50 healthy babies from the Clinical Perinatal Center of Saratov region born at a gestational age of 37-42 weeks with an assessment on the Apgar scale (1952) at the first minute of at least 7 points. After blood sampling, platelet-rich plasma (PRP) was obtained by centrifugation. Results. As a result of the study, 3 forms of platelets were identified: 1. Inactive ones, rounded or more often oval-shaped, in which it is possible to see almost all intracellular structures being typical for platelets; 2. Activated forms that have long processes (pseudopods) and usually have no granules; 3. Transitional forms - apparently, platelets at the initial stages of activation: elements of their open tubular system are already expanding, and this leads to the appearance of vacuoles. However, they do not yet have a distinct rearrangement of the cytoskeleton, and, accordingly, long processes and a significant change in shape. Platelets of newborns are heterogeneous in morphofunctional state: most of them are represented by inactive forms 82(80-85)%, the smaller part includes activated 15(13-17)% and transitional forms 3(2-4)%. Platelet microparticles (PM) are platelet derivatives and are also variable in size and structure. They can be represented by simple vesicles without internal contents, or they can have secretory granules involved in the transmission of signaling information to other platelets and endothelial cells. In our study, several mechanisms of formation and separation of PM from “precursor platelets” were discovered, but their meaning is not yet fully understood and requires further study. Conclusion. This study demonstrates the heterogeneity of the platelet population of newborns associated with their morphofunctional state. For convenience of description, three morphological classes of platelets have been devised. The hypothesis about possible mechanisms of PM formation has been formulated.

Highlights

  • Note: figures A and C show the process of separating platelet microparticles from the platelet without breaching the integrity of the cytoplasmic membrane; figure B shows a defect in the cytolemma at the site of platelet microparticle separation; figure D shows the platelet secretion of small vesicle-shaped platelet microparticles

  • Peripheral blood platelets of healthy newborns were studied by means of transmission electron microscopy (TEM) method

  • Platelet microparticles (PM) are platelet derivatives and are variable in size and structure. They can be represented by simple vesicles without internal contents, or they can have secretory granules involved in the transmission of signaling information to other platelets and endothelial cells

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Summary

МАТЕРИАЛЫ И МЕТОДЫ ИССЛЕДОВАНИЯ

Проведено обследование 50 детей, рожденных без патологии на сроке гестации от 37 до 42 недель. Для исследования осуществлялся забор тромбоцитов из периферической крови. Богатую тромбоцитами плазму (БТП) получали следующим образом: собранную кровь в течение пяти минут центрифугировали на скорости 1200 оборотов в минуту, после чего производился забор надосадка. Полученный в результате этой манипуляции материал готовили для проведения исследования с использованием электронного микроскопа, процедура подготовки проходила в несколько этапов. После этого полученный препарат фиксировали с применением уже 3% раствора ГА, который был приготовлен с использованием того же буфера, при этом время на приготовление раствора составляло двенадцать часов, а температура – плюс четыре градуса Цельсия. 15 мин, после этого аналогичным образом препарат выдерживался в 70% и 90% растворах по 2 раза в течение 15 минут в каждом. После этанола образец три раза помещался в ацетон, время экспозиции составляло 20 мин, а затем – два раза на 30 мин в окись пропилена. Медико-биологические науки грамм Statistica 7.0 for Windows корпорации StatSoft-Russia, Microsoft Office Excel 2016, результаты представлены в виде медианы и квартилей (Ме, Q25, Q75)

РЕЗУЛЬТАТЫ ИССЛЕДОВАНИЯ И ИХ ОБСУЖДЕНИЕ
Gl γ
Gl α
The ratio of the main morphological forms of platelets
ЛИЧНЫЙ ВКЛАД АВТОРОВ
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