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研究血小板添加剂溶液对单采血小板及纤维蛋白网络超微结构的影响。

Investigating the effect of a platelet additive solution on apheresis platelet and fibrin network ultrastructure.

作者信息

Pretorius E, Crookes R, Oberholzer H M, van der Spuy W J

机构信息

Department of Anatomy, University of Pretoria, South Africa.

South African National Blood Service, South Africa.

出版信息

Transfus Apher Sci. 2010 Dec;43(3):347-351. doi: 10.1016/j.transci.2010.10.002. Epub 2010 Oct 30.

Abstract

In thrombotic events and diseases such as cancer, HIV/AIDS, dysfibrinogenaemia, as well as acute incidents (e.g. burn wounds), ultrastructure of platelets and fibrin networks change. In the current study, we compare the ultrastructure of platelets and fibrin networks of apheresis platelets stored in citrated human plasma (CP) and in a first-generation platelet additive solution (PAS) (T-Sol), to that of fresh donor plasma (FP). Eighteen apheresis platelet donors donated platelets on Trima®-Accel™ V5.2 and V5.1 cell separators. Six collections were stored for five days in autologous citrated plasma (CP); six collections were stored in 40% citrated human plasma and 60% PAS solution (CP/PAS) controlled, for the duration of storage, at a constant temperature (22±2°C) with continuous flat-bed agitation; and six collections were stored in conditions uncontrolled for temperature and without continuous agitation. On days 1, 3 and 5, equal volumes of human thrombin were mixed with platelets collected in either CP or CP/PAS to form a coagulum (fibrin network containing platelet aggregates), followed by preparation for scanning electron microscopy. Results were compared with platelets and fibrin networks in FP. Typically, in FP, platelet aggregates with smooth membranes and pseudopodia are seen and fibrin networks arrange to form major, thick fibers and scattered, minor, thin fibers. On day 1, in CP and in all CP/PAS units, platelet ultrastructure compared well to that of FP, although the fibrin fibers were denser, with the minor fibers forming a matted layer over the major fibers. On day 3, in platelet units uncontrolled for temperature and without continuous agitation during storage, some platelet aggregates in CP/PAS showed typical apoptotic morphology, with shrinkage and membrane damage, but comparable fibrin networks were present. On day 5 however, in those units where storage conditions were uncontrolled and where the pH had decreased to below 6.4, no platelet aggregates were seen and fibrin was arranged into short, lumpy masses with no separate major or minor fibrin fibers visible. In those units stored at 22°C with continuous flat-bed agitation, where pH was maintained >7.0, ultrastructure of platelets and fibrin network in CP/PAS was typical and similar to FP and CP at the end of five days of storage. Examining platelet and fibrin network ultrastructure may be useful, in addition to conventional laboratory analysis, in assessing the viability and potential clinical efficacy of platelets for transfusion and could play a role in the evaluation of new generation platelet additive solutions.

摘要

在血栓形成事件以及癌症、艾滋病毒/艾滋病、异常纤维蛋白原血症等疾病和急性事件(如烧伤创面)中,血小板和纤维蛋白网络的超微结构会发生变化。在本研究中,我们将枸橼酸盐人血浆(CP)和第一代血小板添加剂溶液(PAS)(T-Sol)中储存的单采血小板的血小板和纤维蛋白网络超微结构与新鲜供体血浆(FP)的进行比较。18名单采血小板供体在Trima®-Accel™ V5.2和V5.1细胞分离器上捐献血小板。6份采集物在自体枸橼酸盐血浆(CP)中储存5天;6份采集物储存在40%枸橼酸盐人血浆和60%PAS溶液(CP/PAS)中,在储存期间控制在恒定温度(22±2°C)并持续进行平板搅拌;6份采集物在温度不受控制且无持续搅拌的条件下储存。在第1、3和5天,将等量的人凝血酶与在CP或CP/PAS中采集的血小板混合以形成凝块(包含血小板聚集体的纤维蛋白网络),随后制备用于扫描电子显微镜检查。将结果与FP中的血小板和纤维蛋白网络进行比较。通常,在FP中,可以看到具有光滑膜和伪足的血小板聚集体,并且纤维蛋白网络排列形成主要的粗纤维和分散的细纤维。在第1天,在CP和所有CP/PAS单位中,血小板超微结构与FP的相比良好,尽管纤维蛋白纤维更密集,细纤维在粗纤维上形成一层缠结层。在第3天,在储存期间温度不受控制且无持续搅拌的血小板单位中,CP/PAS中的一些血小板聚集体呈现典型的凋亡形态,伴有收缩和膜损伤,但存在可比的纤维蛋白网络。然而,在第5天,在储存条件不受控制且pH降至6.4以下的那些单位中,未观察到血小板聚集体,并且纤维蛋白排列成短的块状物,没有可见的单独粗纤维或细纤维。在以22°C持续平板搅拌储存的那些单位中,pH维持>7.0,CP/PAS中血小板和纤维蛋白网络的超微结构在储存5天后是典型的且与FP和CP相似。除了常规实验室分析外,检查血小板和纤维蛋白网络超微结构可能有助于评估用于输血的血小板的活力和潜在临床疗效,并且可能在评估新一代血小板添加剂溶液中发挥作用。

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