Table 2.

Studies^a investigating endothelial integrity through plasma exposition in HS conditions.

Reference	Experimental models	Types of plasma	Main results
Pati et al. (39)	Studies on HUPECs monolayers (hypoxia-induced permeability) with assessment of EC permeability (FITC-Dextran) after FFP treatment, comparing FFP stored for 0 vs FFP stored for 5 days In vivo studies on rat model of HS for testing capacity of FFP (comparing FFP stored for 0 vs for 5 days) to restore MAP	Human FFP (ABO blood types, same donor or pooled from three donors, thawed-aliquoted-stored at 4°C for 0 or 5 days before use)	Day 0 FFP inhibits EC permeability; day 5 FFP demonstrated a diminished capacity to inhibit EC permeability Day 0 FFP, but not day 5 FFP, restores blood pressure to baseline
Kozar et al. (40)	Studies on a rat model of HS, comparing effect of LR vs fresh plasma resuscitation with assessment of endothelial glycocalyx on mesenteric vessels (electronic microscopy), relative expression level of syndecan-1 (QRT RT PCR) and cell surface expression of syndecan-1 (immunostaining) in lung tissue	Fresh plasma (not otherwise specified)	Glycocalyx is partially restored by plasma resuscitation Syndecan-1 expression in lung is enhanced by plasma Lung injury is lessened by plasma resuscitation
Haywood-Watson et al. (86)	Studies on HUVECs monolayers (hypoxia-induced permeability) with assessment of VE-cadherin and syndecan-1 expression (immunofluorescence), topographical properties (AFM), permeability (FITC-Dextran) after LR vs FFP treatment Patients admitted to ICU for shock, resuscitation with plasma, syndecan-1 and cytokines measurements	FFP (not otherwise specified)	Vascular integrity is disrupted by shock but mitigated by FFP FFP hastens syndecan-1 restoration compared to LR Injured patients in shock shed syndecan-1; syndecan-1 correlates with specific inflammatory cytokines
Torres et al. (42)	Studies on a rat HS models comparing effect of LR/HS vs fresh plasma resuscitation with studies on blood samples (including thromboelastometry) and on endothelium (glycocalyx thickness measurements by fluorescent dye-exclusion method)	FFP defined as plasma frozen within 6–8 h of collection and stored at −20°C, prepared by separation form whole blood collected on donor rats	Restoration of coagulation function by a small-volume resuscitation with FFP in contrast to resuscitation with LR/HS groups
Peng et al. (41)	Studies on HUPECs monolayers (VEGF-A165-induced permeability) with assessment of EC permeability (TEER/ECIS and FITC-Dextran) and leukocyte-endothelial binding Mouse model of HS and trauma comparing effect of LR vs FFP resuscitation with in vivo studies (MAP monitoring, measurement of syndecan-1 in plasma) and in vitro studies on harvested lungs: vascular permeability (intravenous fluorescent dye extravasation), infiltration of neutrophils (MPO immunofluorescence staining and activity), syndecan-1 detection (anti-syndecan-1 antibody)	Human FFP used in both in vitro and in vivo studies (frozen within 8 h after donation, kept frozen until the day of experiment and used within 1–2 h of thaw)	In HUPECs monolayers, FFP compared with LR reduces pulmonary endothelial hyper-permaeability and leukocyte binding In mouse HS models, FFP and LR similarly restore MAP. FFP mitigates lung hyper-permeability, reduces lung inflammation, increases lung syndecan-1, and reduces syndecan-1 shedding compared with LR resuscitation
Wataha et al. (44)	Studies on HUVECs and PECs monolayers (VEGF-A165-induced permeability) comparing effect of FFP, SD-FFP, SDP (controls: LR/HS) with assessment of EC permeability (FITC-Dextran), WBC binding assay (fluorescent labeling), surface adhesion molecules/integrin expression (flow cytometry) and VE-cadherin/β-catenin mobilization to cell surface (staining)	Human FFP (frozen at −20°C, thawed at 37°C and used on day 0–1 of thaw)SDP defined as pooled liquid plasma that has been dehydrated by means of spray drying and reconstituted citric acid and monobasic sodium phosphate (SD-FFP being the starting material)	FFP, SD-FFP, and SPD equivalently inhibit vascular permeability, ensures EC adherens junctions integrity and endothelial WBC binding Lack of difference between FFP and SD-FFP and between SD-FFP and SDP indicating that solvent-detergent treatment and spray drying do not affect the ability of plasma product to modulate endothelial function
Potter et al. (45)	Studies on HUVECs monolayers (VEGF-A165-induced permeability), comparing FFP and SDP (controls: LR) by testing endothelial permeability (TEER/ECIS), cytokine production in EC and gene expression Mouse model of HS comparing FFP and SDP (controls: LR) with in vivo studies (MAP and BE monitoring) and measurement of EC adherent junctions stability (immunofluorescence and histological staining) on harvested lungs	FFP obtained from human donors plasma by apheresis collection, used freshly thawed (same day of thaw)SDP from multidonor plasma (more than 150 type AB donors)	On HUVECs monolayers, FFP and SDP decrease endothelial permeability, induce similar patterns of gene expression and cytokines production in EC In mouse HS models, SDP and FFP equivalently correct MAP and BE, reduce pulmonary vascular leak, equivalently inhibit leukocyte infiltration and breakdown of endothelial adherens and tight junctions
Torres Filho et al. (90)	Rat model of HS for studying quantitatively the relationship between plasma biomarkers and changes in microvascular parameters, including glycocalyx thickness after resuscitation with FWB, PRBC, FFP, 5% albumin, or crystalloids (RL, NS, and HTS)	FWB (3.2% citrate, stored at 4°C, used with 24 h), PRBC (used within 48 h), and FFP (frozen within 6–8 h of collection, stored at −80°C for up to 1 year) all from donor rats	Changes in glycocalyx thickness (and microvascular permeability) negatively (positively) correlated with changes in plasma levels of syndecan-1 and heparane sulfate FWB and FFP, but neither colloid or crystalloid resuscitation, support vascular stabilization by reconstitution of the endothelia glycocalyx after HS
Diebel et al. (91)	HUVEC lined microfluidics model for studying endothelial cell activation/injury and glycocalyx barrier function after simulation of HS by treatment with epinephrine and hypoxia reoxygenation	5% human plasma perfused immediately following treatment or after a 3 h delay	“Early” plasma mitigates glycocalyx degradation and inflammatory prothrombotic endothelial response
Pati et al. (92)	Studies on HUVECs monolayers (VEGF-A165-induced permeability), comparing FFP and LP (controls: LR or no treatment) by testing EC permeability (FITC-Dextran), EC resistance (TEER/ECIS), VE-cadherin/β-catenin mobilization to cell surface (staining), leukocyte-binding (fluorescent labeling) Mouse model of HS comparing FFP and LP (controls: LR or no treatment) with assessment of inflammation (MPO staining), vascular permeability (dye extravasation) and tissue edema (wet-to-dry weight ratio)	Human FFP (male donors O+) thawed and used freshly (day 0 of thaw) LP defined as lyophilized plasma (male O+) reconstituted in buffer	On HUVECs monolayers, FFP and LP decrease endothelial permeability, preserve EC adherens junctions, attenuate EC-leukocyte-binding In mouse HS models, LP and FFP reduce pulmonary vascular permeability, edema, and inflammation

AFM, atomic force microscopy; BE, base excess; EC, endothelial cell; ECIS, electric cell-substrate impedance system; FFP, fresh frozen plasma; FITC, fluorescein isothiocyanate-conjugated; FWB, fresh whole blood; HES, hydroxyethyl starch; HS, hemorrhagic shock; HTS, hypertonic (3%) sodium chloride; HUPEC, human pulmonary endothelial cell; HUVEC, human umbilical vein endothelial cell; LP, lyophilized plasma; LR, lactated ringers; MAP, mean arterial pressure; NS, normal saline; PEC, pulmonary endothelial cells; PRBC, packed red blood cells; QRT RT PCR, quantitative real-time reverse-transcription polymerase chain reaction; SD, solvent detergent; SDP, spray-dried plasma; TEER, trans-endothelial electrical resistance; VE-cadherin, vascular endothelial cadherin; WBC, white blood cell.

^aStudies identified by searching the terms “glycocalyx, haemorrhagic shock, plasma” on PubMed and secondary references.