Platelet transmission electron microscopy for the assessment of poor biological response to antiplatelet agent: pilot descriptive and prospective study - ELECTROSTROKE

Abstract

Introduction: Ischaemic stroke is the leading cause of adult disability. Thus, a strategy based on an efficient antiplatelet therapy has been developed. The monitoring of antiplatelet therapy is now limited to high risk and poor prognosis patients. Indeed, the biological monitoring of the antiplatelet therapy with available platelet function assays do not provide a global integrative approach. Platelet transmission electron microscopy, recently validated for assessing distinct ultrastructural abnormalities is a reliable morphological platelet structural analysis tool which could be used to collect all the ultrastructural platelet characteristics and assess the degree of activation of platelets.

Methods and analysis: Our pilot prospective and descriptive study will include 50 consecutive patients hospitalized for an ischaemic stroke. We expect to identify ultrastructural characteristics that will be correlated with the degree of platelet activation to guide clinicians in decision making regarding the antiplatelet therapy strategy.

Ethics and dissemination: The French Ethics Committee (comité de protection des personnes d'Ile-de-France VII) approved the information notice that will be given to participants and the protocol of this trial (protocol No 21-031).The results of the trial will be disseminated through scientific publications.

Trial registration number: NCT05004233.

Keywords: Stroke; Stroke medicine; Vascular medicine.

Figure 1

Outline of the trial. TEM, transmission electron microscopy.

Figure 2

Process of the platelet samples: collection of the citrated blood, fixation of the whole blood, preparation of the platelet rich plasma (PRP), transport of the samples to the electron microscopy facilities, centrifugation of the PRP, removal of the supernatant, washing step, post-fixation step, washing steps, dehydration steps, epoxy embedding, 70 nm ultrathin sections, contrast with Uranyless and lead citrate, transmission electron microscopy observations, 2D and 3D analysis, description of ultrastructural characteristics and preliminary evaluation of low biological responsiveness to an antiplatelet therapy. 2D, two dimensions; 3D, three dimensions.