The Structural Stability of the Endothelial Glycocalyx after Enzymatic Removal of Glycosaminoglycans

Zeng, Ye; Ebong, Eno E.; Fu, Bingmei M.; Tarbell, John M.

doi:10.1371/journal.pone.0043168

Cited by 105 publications

(111 citation statements)

References 56 publications

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“…We previously reported on the persisting structural stability of the whole glycocalyx, even after the enzymatic removal of individual glycosaminoglycans. 38 However, the results from the current heparinase III experiment produced a decrease in total glycocalyx thickness but statistically negligible change in the continuity of the total glycocalyx. The discrepancy between our previous and presently reported measurements may have been due to the treatment time scale.…”

Section: Resultsmentioning

confidence: 53%

“…The dimensions of total glycocalyx (BSA) and the HS subcomponent measured were within the range of previously reported values for rodent cell cultures and animal models using the same fixation and microscopy approach. 37,38,45 Therefore, these results provide strong indication that the untreated RFPEC glycocalyx is healthy; owing to its substantial thickness and continuity, the RFPEC glycocalyx mesh is expected to serve as a robust, molecular sieve as seen in the body.…”

Section: Resultsmentioning

confidence: 82%

“…RFPECs were selected for their ability to naturally express a robust glycocalyx. 38 These ECs were cultured in tissue culture flasks at 37°C and 5% CO 2 in a humidified environment with Dulbecco's Modified Eagle's Medium (DMEM) supplemented with 10% fetal bovine serum (FBS) and 1% penicillin-streptomycin, all purchased from Thermo Fisher Scientific (Waltham, MA, USA). …”

Section: Materials and Methods Rfpec Culturementioning

confidence: 99%

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Endothelial glycocalyx conditions influence nanoparticle uptake for passive targeting

Cheng¹,

Kumar²,

Sridhar³

et al. 2016

IJN

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Cardiovascular diseases are facilitated by endothelial cell (EC) dysfunction and coincide with EC glycocalyx coat shedding. These diseases may be prevented by delivering medications to affected vascular regions using circulating nanoparticle (NP) drug carriers. The objective of the present study was to observe how the delivery of 10 nm polyethylene glycol-coated gold NPs (PEG-AuNP) to ECs is impacted by glycocalyx structure on the EC surface. Rat fat pad endothelial cells were chosen for their robust glycocalyx, verified by fluorescent immunolabeling of adsorbed albumin and integrated heparan sulfate (HS) chains. Confocal fluorescent imaging revealed a ~3 µm thick glycocalyx layer, covering 75% of the ECs and containing abundant HS. This healthy glycocalyx hindered the uptake of PEG-AuNP as expected because glycocalyx pores are typically 7 nm wide. Additional glycocalyx models tested included: a collapsed glycocalyx obtained by culturing cells in reduced protein media, a degraded glycocalyx obtained by applying heparinase III enzyme to specifically cleave HS, and a recovered glycocalyx obtained by supplementing exogenous HS into the media after enzyme degradation. The collapsed glycocalyx waŝ2 µm thick with unchanged EC coverage and sustained HS content. The degraded glycocalyx showed similar changes in EC thickness and coverage but its HS thickness was reduced to 0.7 µm and spanned only 10% of the original EC surface. Both dysfunctional models retained six- to sevenfold more PEG-AuNP compared to the healthy glycocalyx. The collapsed glycocalyx permitted NPs to cross the glycocalyx into intracellular spaces, whereas the degraded glycocalyx trapped the PEG-AuNP within the glycocalyx. The repaired glycocalyx model partially restored HS thickness to 1.2 µm and 44% coverage of the ECs, but it was able to reverse the NP uptake back to baseline levels. In summary, this study showed that the glycocalyx structure is critical for NP uptake by ECs and may serve as a passive pathway for delivering NPs to dysfunctional ECs.

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Section: Resultsmentioning

confidence: 53%

Section: Resultsmentioning

confidence: 82%

Section: Materials and Methods Rfpec Culturementioning

confidence: 99%

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Endothelial glycocalyx conditions influence nanoparticle uptake for passive targeting

Cheng¹,

Kumar²,

Sridhar³

et al. 2016

IJN

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“…While there is considerable debate about the thickness of the glycocalyx both in vitro and in vivo (reviewed in Ebong et al [67]), the choice of 500 nm is a reasonable estimate of the thickness of the denser inner layer that is mechanically significant. The thickness of glycocalyx has been shown to be fairly uniform on cultured cells which have not been exposed to shear stress [68,69]. Bai & Wang [25] investigated the spatial distribution and temporal development of the glycocalyx on cultured human umbilical vein ECs (HUVECs).…”

Section: Geometric Modelmentioning

confidence: 99%

Shear-induced force transmission in a multicomponent, multicell model of the endothelium

Dabagh

Jalali

Butler

et al. 2014

J. R. Soc. Interface.

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Haemodynamic forces applied at the apical surface of vascular endothelial cells (ECs) provide the mechanical signals at intracellular organelles and through the inter-connected cellular network. The objective of this study is to quantify the intracellular and intercellular stresses in a confluent vascular EC monolayer. A novel three-dimensional, multiscale and multicomponent model of focally adhered ECs is developed to account for the role of potential mechanosensors (glycocalyx layer, actin cortical layer, nucleus, cytoskeleton, focal adhesions (FAs) and adherens junctions (ADJs)) in mechanotransmission and EC deformation. The overriding issue addressed is the stress amplification in these regions, which may play a role in subcellular localization of mechanotransmission. The model predicts that the stresses are amplified 250-600-fold over apical values at ADJs and 175-200-fold at FAs for ECs exposed to a mean shear stress of 10 dyne cm 22 . Estimates of forces per molecule in the cell attachment points to the external cellular matrix and cell-cell adhesion points are of the order of 8 pN at FAs and as high as 3 pN at ADJs, suggesting that direct force-induced mechanotransmission by single molecules is possible in both. The maximum deformation of an EC in the monolayer is calculated as 400 nm in response to a mean shear stress of 1 Pa applied over the EC surface which is in accord with measurements. The model also predicts that the magnitude of the cell-cell junction inclination angle is independent of the cytoskeleton and glycocalyx. The inclination angle of the cell-cell junction is calculated to be 6.68 in an EC monolayer, which is somewhat below the measured value (9.98) reported previously for ECs subjected to 1.6 Pa shear stress for 30 min. The present model is able, for the first time, to cross the boundaries between different length scales in order to provide a global view of potential locations of mechanotransmission.

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“…In the vascular remodeling due to AS, the structural and functional adaptive changes in vessels adapt to the mechanical (such as shear stress) and chemical (such as vasoactive mediators and cytokines) microenvironments, which involve various cell activities such as cell phenotype conversion [9] and remodeling of the extracellular matrix and endothelial glycocalyx [10][11][12][13][14]. It is well-known that flow patterns with shear stress magnitude are changed with development of AS [15].…”

Section: Introductionmentioning

confidence: 99%

Endothelial Glycocalyx: Novel Insight into Atherosclerosis

Zeng¹

2017

J. Biomed

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The evidence shows that endothelial glycocalyx exerts an important role in inflammation and atherosclerosis. We previously demonstrated that hydrodynamic force such as shear stress induces the remodeling of the major component of glycocalyx including glypican-1 with attached heparan sulfate, and that sphingosine-1-phosphate (S1P) protects the glycocalyx against syndecan-1 ectodomain shedding and induces the synthesis of HS. Actually, both shear stress and S1P imposed significant influence on inflammatory disease such as atherosclerosis. Therefore, it can be concluded that glycocalyx is a critical signaling platform for determining the fate of endothelial cells and atherosclerosis. This review integrated our current understanding of shear stress, S1P and glycocalyx, and also provided new insight/issues into the role of the glycocalyx in the process of forming and developing atherosclerosis.

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The Structural Stability of the Endothelial Glycocalyx after Enzymatic Removal of Glycosaminoglycans

Cited by 105 publications

References 56 publications

Endothelial glycocalyx conditions influence nanoparticle uptake for passive targeting

Endothelial glycocalyx conditions influence nanoparticle uptake for passive targeting

Shear-induced force transmission in a multicomponent, multicell model of the endothelium

Endothelial Glycocalyx: Novel Insight into Atherosclerosis

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