Hongyan Kang

Vascular smooth muscle cell glycocalyx modulates shear-induced proliferation, migration, and NO production responses

The endothelial cell glycocalyx, a structure coating the luminal surface of the vascular endothelium, and its related mechanotransduction have been studied by many over the last decade. However, the role of vascular smooth muscle cells (SMCs) glycocalyx in cell mechanotransduction has triggered little attention. This study addressed the role of heparan sulfate proteoglycans (HSPGs), a major component of the glycocalyx, in the shear-induced proliferation, migration, and nitric oxide (NO) production of the rat aortic smooth muscle cells (RASMCs). A parallel plate flow chamber and a peristaltic pump were employed to expose RASMC monolayers to a physiological level of shear stress (12 dyn/cm(2)). Heparinase III (Hep.III) was applied to selectively degrade heparan sulfate on the SMC surface. Cell proliferation, migration, and NO production rates were determined and compared among the following four groups of cells: 1) untreated with no flow, 2) Hep.III treatment with no flow, 3) untreated with flow of 12 dyn/cm(2) exposure, and 4) Hep.III treatment with flow of 12 dyn/cm(2) exposure. It was observed that flow-induced shear stress significantly suppressed SMC proliferation and migration, whereas cells preferred to aligning along the direction of flow and NO production were enhanced substantially. However, those responses were not found in the cells with Hep.III treatment. Under flow condition, the heparinase III-treated cells remained randomly oriented and proliferated as if there were no flow presence. Disruption of HSPG also enhanced wound closure and inhibited shear-induced NO production significantly. This study suggests that HSPG may play a pivotal role in mechanotransduction of SMCs.

Effect of swirling flow on the uptakes of native and oxidized LDLs in a straight segment of the rabbit thoracic aorta

To elucidate the physiological significance of the spiral flow in the arterial system from the viewpoint of atherogenic lipid transport, an ex vivo experimental comparative study was designed to investigate the effect of swirling flow on the distribution of native 3,3′-dioctadecylindocarbocyanine-low-density lipoprotiens (DiI-LDL) and DiI-ox-LDL uptakes by segments of the rabbit thoracic aorta. The experimental results showed that when compared with the normal flow, the swirling flow generated in the test arteries significantly reduced the DiI-LDL and DiI-ox-LDL uptakes by the arterial walls. The results also showed that the values of DiI-ox-LDL uptake were higher than those of DiI-LDL uptake at the same sample position in both the normal flow group and the swirling flow group. Most interestingly, the experimental results found that the percentage increase in DiI-ox-LDL uptake was much larger than that in DiI-LDL uptake when the perfusion duration increased from 3 to 24 h. In conclusion, the present study substantiated the hypothesis that the spiral flow in the arterial system plays a beneficial role in protecting the arterial wall from atherogenesis. Meanwhile, it supported the concept that the receptor-mediated bindings of LDL uptake, the barrier function of the arterial endothelial linings and the mass transport phenomenon of LDL concentration polarization are all involved in the infiltration/accumulation of atherogenic lipids within the arterial wall.

3,3'-Dioctadecylindocarbocyanine-low-density lipoprotein uptake and flow patterns in the rabbit aorta-iliac bifurcation under three perfusion flow conditions.

The aim of this study was to elucidate which of the following two factors plays a more important role in the localization of atherogenesis: the barrier function of the arterial endothelium modulated by wall shear stress or flow-dependent low-density lipoprotein (LDL) concentration at the blood/wall interface. To determine this, the rabbit aorto-iliac bifurcation was perfused with 3,3′-dioctadecylindocarbocyanine (DiI)-LDL solution under three different flow conditions: (i) forward flow (perfused in the in vivo flow direction); (ii) backward flow (perfused in a reversed flow direction); and (iii) static group (no flow). The results showed that there was a peak in the curve of DiI-LDL uptake distribution along the lateral wall of the bifurcation for all three groups, which was located in the branching areas where the endothelial cells were round and polygonal with no preferred orientation. Nevertheless, the peak of the forward flow group was much sharper than those of the other two groups. The overall DiI-LDL uptake was the highest for the static group. The present experimental study supports the concept that both the barrier function of the endothelium modulated by wall shear stress and the mass transport phenomenon of LDL concentration polarization are involved in the infiltration/accumulation of atherogenic lipids within the arterial wall. Nevertheless, the latter might play a larger role in the localization of atherogenesis.

Compositional or charge density modification of the endothelial glycocalyx accelerates flow-dependent concentration polarization of low-density lipoproteins

We hypothesized that diminished endothelial glycocalyx (GCX) at atherosclerotic lesion-prone sites accelerates flow-dependent concentration polarization of low-density lipoproteins (LDLs) at the luminal surface, and in turn contributes to vulnerability of these sites to atherosclerosis. A parallel plate flow chamber was applied to expose cultured endothelial monolayers to three different levels of shear stress (3, 12, 20 dyn/cm2). Heparinase III (Hep.III) was employed to degrade heparan sulfate proteoglycans selectively and 3-(N-morpholino) propanesulfonic acid-buffered physiological salt solutions (MOPS-PSS) were used at either normal ionic strength (Normal-MOPS), low ionic strength (LO-MOPS) or high ionic strength (HI-MOPS) to modify the effective charge density of the endothelial GCX. Water filtration velocity (V w ) across the endothelial monolayer, the luminal concentration of LDLs (C w ) and the uptake of LDLs by endothelial cells were measured and compared among the following five groups of cells: (1) Control; (2) Hep.III treatment; (3) LO-MOPS; (4) Normal-MOPS; and (5) HI-MOPS. The results obtained substantiated the aforementioned hypothesis and demonstrated that compositional or charge density modification of the endothelial GCX facilitated water filtration across the endothelium, enhanced the accumulation of LDLs on the luminal surface and increased the uptake of LDLs by endothelial cells, therefore contributing to atherogenesis.

A Potential Gravity-Sensing Role of Vascular Smooth Muscle Cell Glycocalyx in Altered Gravitational Stimulation

Previously, we have shown that vascular smooth muscle cells (VSMCs) exhibit varied physiological responses when exposed to altered gravitational conditions. In the present study, we focused on elucidating whether the cell surface glycocalyx could be a potential gravity sensor. For this purpose, a roller culture apparatus was used with the intent to provide altered gravitational conditions to cultured rat aortic smooth muscle cells (RASMCs). Heparinase III (Hep.III) was applied to degrade cell surface heparan sulfate proteoglycans (HSPG) selectively. Sodium chlorate was used to suppress new synthesis of HSPG. Glycocalyx remodeling, nitric oxide synthase (NOS) activation, and F-actin expression induced by gravity alteration were assessed by flow cytometry, reverse transcription polymerase chain reaction (RT-PCR), and Western blot. Results indicate that the exposure of cultured RASMCs to altered gravitational conditions led to a reduction in cell surface HSPG content and the activation of NOS. It also down-regulated the expression of glypican-1, constitutive NOS (NOSI and NOSIII), and F-actin. On the other hand, Hep.III followed by sodium chlorate treatment of HSPG attenuated the aforementioned NOS and F-actin modulation under altered gravitational conditions. All these findings suggest that the glycocalyx, and HSPG in particular, may be an important sensor of gravitational changes. This may play an important role in the regulation of NOS activation, F-actin modulation, and HSPG remodeling in VSMCs.

High glucose–induced endothelial progenitor cell dysfunction:

Vascular complications contribute significantly to morbidity and mortality of diabetes mellitus. The primary cause of vascular complications in diabetes mellitus is hyperglycaemia, associated with endothelial dysfunction and impaired neovascularization. Circulating endothelial progenitor cells was shown to play important roles in vascular repair and promoting neovascularization. In this review, we will demonstrate the individual effect of high glucose on endothelial progenitor cells. Endothelial progenitor cells isolated from healthy subjects exposed to high glucose conditions or endothelial progenitor cells isolated from diabetic patients exhibit reduced number of endothelial cell colony forming units, impaired abilities of differentiation, proliferation, adhesion and migration, tubulization, secretion, mobilization and homing, whereas enhanced senescence. Increased production of reactive oxygen species by the mitochondria seems to play a crucial role in high glucose–induced endothelial progenitor cells deficit. Later, we will review the agents that might be used to alleviate dysfunction of endothelial progenitor cells induced by high glucose. The conclusions are that the relationship between hyperglycaemia and endothelial progenitor cells dysfunction is only beginning to be recognized, and future studies should pay more attention to the haemodynamic environment of endothelial progenitor cells and ageing factors to discover novel treatment agents.

Vascular smooth muscle cell glycocalyx mediates shear stress-induced contractile responses via a Rho kinase (ROCK)-myosin light chain phosphatase (MLCP) pathway

The vascular smooth muscle cells (VSMCs) are exposed to interstitial flow induced shear stress that may be sensed by the surface glycocalyx, a surface layer composed primarily of proteoglycans and glycoproteins, to mediate cell contraction during the myogenic response. We, therefore, attempted to elucidate the signal pathway of the glycocalyx mechanotransduction in shear stress regulated SMC contraction. Human umbilical vein SMCs (HUVSMCs) deprived of serum for 3–4 days were exposed to a step increase (0 to 20 dyn/cm2) in shear stress in a parallel plate flow chamber, and reduction in the cell area was quantified as contraction. The expressions of Rho kinase (ROCK) and its downstream signal molecules, the myosin-binding subunit of myosin phosphatase (MYPT) and the myosin light chain 2 (MLC2), were evaluated. Results showed that the exposure of HUVSMCs to shear stress for 30 min induced cell contraction significantly, which was accompanied by ROCK1 up-regulation, re-distribution, as well as MYPT1 and MLC activation. However, these shear induced phenomenon could be completely abolished by heparinase III or Y-27632 pre-treatment. These results indicate shear stress induced VSMC contraction was mediated by cell surface glycocalyx via a ROCK-MLC phosphatase (MLCP) pathway, providing evidence of the glycocalyx mechanotransduction in myogenic response.

Numerical simulation of haemodynamics and low-density lipoprotein transport in the rabbit aorta and their correlation with atherosclerotic plaque thickness

Two mechanisms of shear stress and mass transport have been recognized to play an important role in the development of localized atherosclerosis. However, their relationship and roles in atherogenesis are still obscure. It is necessary to investigate quantitatively the correlation among low-density lipoproteins (LDL) transport, haemodynamic parameters and plaque thickness. We simulated blood flow and LDL transport in rabbit aorta using computational fluid dynamics and evaluated plaque thickness in the aorta of a high-fat-diet rabbit. The numerical results show that regions with high luminal LDL concentration tend to have severely negative haemodynamic environments (HEs). However, for regions with moderately and slightly high luminal LDL concentration, the relationship between LDL concentration and the above haemodynamic indicators is not clear cut. Point-by-point correlation with experimental results indicates that severe atherosclerotic plaque corresponds to high LDL concentration and seriously negative HEs, less severe atherosclerotic plaque is related to either moderately high LDL concentration or moderately negative HEs, and there is almost no atherosclerotic plaque in regions with both low LDL concentration and positive HEs. In conclusion, LDL distribution is closely linked to blood flow transport, and the synergetic effects of luminal surface LDL concentration and wall shear stress-based haemodynamic indicators may determine plaque thickness.

Influence of catheter insertion on the hemodynamic environment in coronary arteries.

Intravascular stenting is one of the most commonly used treatments to restore the vascular lumen and flow conditions, while perioperative complications such as thrombosis and restenosis are still nagging for patients. As the catheter with crimped stent and folded balloon is directly advanced through coronary artery during surgery, it is destined to cause interference as well as obstructive effect on blood flow. We wonder how the hemodynamic environment would be disturbed and weather these disturbances cause susceptible factors for those complications. Therefore, a realistic three-dimensional model of left coronary artery was reconstructed and blood flow patterns were numerically simulated at seven different stages in the catheter insertion process. The results revealed that the wall shear stress (WSS) and velocity in left anterior descending (LAD) were both significantly increased after catheter inserted into LAD. Besides, the WSS on the catheter, especially at the ending of the catheter, was also at high level. Compared with the condition before catheter inserted, the endothelial cells of LAD was exposed to high-WSS condition and the risk of platelet aggregation in blood flow was increased. These influences may make coronary arteries more vulnerable for perioperative complications.

Bioinspired helical graft with taper to enhance helical flow

Helical flow has been introduced to improve the hemodynamic performance of vascular devices such as arterial grafts, stents and arteriovenous shunts to overcome the flow induced thrombus formation and intimal hyperplasia. However, the quite low intensity of helical flow in the existing devices may limit their function. To obtain desirably high intensity, inspired by the helical flow and tapered configuration of the arterial system, we proposed a new conceptual design of the medical devices, which take the form of a tapered helical shape. We demonstrated its effectiveness in arterial grafts by numerically comparing the hemodynamic performance of helical grafts with different smooth tapers. The results show that the helicity density quantifying the helical flow enlarges sharply with the increase of the taper under both steady and pulsatile flow conditions. Moreover, the amplified helical flow induced by the taper would lead to highly elevated wall shear stress, remarkably reduced oscillating shear index and relative residence time at both the grafts and the anastomosis of the host vessel. The present findings therefore indicated that the new helical graft with taper would significantly enhance the helical flow in the grafts and host vessel, which may reduce the possibility of thrombus formation in the graft and intimal hyperplasia in the host vessel and hence improve the graft patency. In addition, the concept of helical shape with taper may also be applied to design arterial stents and arteriovenous shunts to obtain better hemodynamic performance.