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The aortic valve was long considered a passive structure that opens and closes in response to changes in transvalvular pressure. Recent evidence suggests that the aortic valve performs highly sophisticated functions as a result of its unique microscopic structure. These functions allow it to adapt to its hemodynamic and mechanical environment. Understanding the cellular and molecular mechanisms involved in normal valve physiology is essential to elucidate the mechanisms behind valve disease. We here review the structure and developmental biology of aortic valves; we examine the role of its cellular parts in regulating its function and describe potential pathophysiological and clinical implications.

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REVIEW ARTICLE

Cellular regulation of the structure and function

of aortic valves

Ismail El-Hamamsy, Adrian H Chester, Magdi H Yacoub *

Harefield Heart Science Center, National Heart and Lung Institute, Imperial College London, United Kingdom

KEYWORDS

Aortic valve;

Endothelium;

Mechanotransduction;

Aortic valve calcification;

Interstitial cells

Abstract The aortic valve was long considered a passive structure that opens and closes in response to changes in transvalvular pressure Recent evidence suggests that the aortic valve per-forms highly sophisticated functions as a result of its unique microscopic structure These functions allow it to adapt to its hemodynamic and mechanical environment Understanding the cellular and molecular mechanisms involved in normal valve physiology is essential to elucidate the mechanisms behind valve disease We here review the structure and developmental biology of aortic valves; we examine the role of its cellular parts in regulating its function and describe potential pathophysio-logical and clinical implications

ª 2009 University of Cairo All rights reserved.

Introduction

The aortic valve lies at the junction between the left ventricle and

the aorta It opens and closes >100.000 times daily For a long

time, the aortic valve was believed to be a passive structure that

opens and closes in response to changes in transvalvular

pres-sures Although this is partly true, recent evidence clearly

dem-onstrates that the aortic valve has a tightly regulated and highly conserved architecture which allows it to perform sophisticated functions, in turn affecting systolic blood flow, ventricular workload and coronary blood flow, among other things In addition, the cellular components of the valve play an important role in maintaining normal valve architecture and composition Dysregulation in one of the elements can lead to dysfunction and dysmorphic changes of the valve In this review, we will dis-cuss the unique structure of the aortic valve We will then focus

on the role of aortic valve endothelial cells in maintaining valve structure and function and their possible role in calcified aortic valve disease, with an emphasis on clinical implications Aortic valve structure

Macroscopic structure

The aortic valve mechanism is composed of four inter-related component parts which form a functional unit: the aortic annulus, the aortic cusps, the sinuses of Valsalva and the sin-otubular junction The normal aortic valve is composed of

* Corresponding author Address: Harefield Heart Science Center,

Harefield Hospital, Hill End Road, Harefield, Middlesex, United

Kingdom Fax: +44 1895 828 900.

E-mail address: m.yacoub@imperial.ac.uk (M.H Yacoub).

2090-1232 ª 2009 University of Cairo All rights reserved Peer review

under responsibility of University of Cairo.

2090-1232 ª 2009 University of Cairo All rights reserved Peer review

under responsibility of University of Cairo.

Production and hosting by Elsevier

University of Cairo

Journal of Advanced Research

doi:10.1016/j.jare.2010.02.007

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three semi-lunar cusps that are attached to a crown-shaped

annulus at their base Adequate and coordinated opening of

the aortic valve is essential to ensure unobstructed laminar

blood flow from the left ventricle and decrease ventricular

workload in systole [1–4] Similarly, proper closure of the

cusps in diastole preserves the shape of the root and

contrib-utes to the creation of vortices in the sinuses of Valsalva, an

important determinant of adequate coronary blood flow in

diastole and in systole [5,6] The aortic valve lies in a unique

hemodynamic and mechanical environment exposing the cusps

to a wide range of stresses during the cardiac cycle, which

range from pressure, to tension and bending forces Although

aortic valve cusps are extremely thin structures, yet both sides

of the valve are exposed to different stresses, in particular

shear stress which is a major stimulus for valve endothelial

cells (VECs) The ventricular side of the cusps is exposed to

high-shear stress due to a systolic, high velocity, laminar blood

flow, whereas the aortic side of the cusps is exposed to

low-shear stress secondary to diastolic, low velocity, disturbed

blood flow VECs have the ability to sense changes in shear

stress and to translate these mechanical stimuli into biological

responses (mechanotransduction)[7] Previous studies focusing

on the vascular endothelium show that different patterns of

flow can greatly influence the response of underlying tissues

through activation of specific mechanotransduction pathways

in the endothelium, which can lead to structural changes at

the level of the vessel wall

Cellular structure

In addition to the VECs which line both sides of the cusp, the

body of the aortic valve is composed of valve interstitial cells

(VICs) lying within the extracellular matrix (ECM) (Fig 1)

VICs are composed of different cell types, namely fibroblasts,

smooth muscle cells and myofibroblasts [8] Smooth muscle

cells can exhibit both secretory and contractile properties

Along with the fibroblasts, these secretory properties are

responsible for generation, maintenance and repair of the

ECM which is mainly composed of elastin, collagen and

prote-oglycans[9] The three-dimensional microscopic architecture of

the aortic valve cusps is highly preserved between species and consists of three layers distinguishable by their chemical com-position and mechanical properties[10](Fig 1) On the aortic side lies the fibrosa, a layer rich in collagen fibers which pro-vides most of the tensile strength to the valve On the ventricu-lar side is the ventricuventricu-laris which is rich in elastin fibers, thus providing elasticity to the aortic valve Between these two layers

is the spongiosa, which represents about 60–70% of the thick-ness of the cusp and is primarily composed of proteoglycans Proteoglycans are highly hydrated, thus acting as ‘‘shock absorbers’’ during the different parts of the cardiac cycle

In addition to this basic structure of aortic valves, a popu-lation of resident stem cells lying within the cusps has recently been recognized[11] They appear to originate from the mobi-lization of hematopoietic-derived stem cells towards cardiac valves [12] Their role is not yet fully understood but they are thought to contribute to valve repair, cell regeneration,

as well as participating in valve calcification in some disease states as will be discussed later Although aortic valves are avascular structures which extract their nutrients by extraction from surrounding blood, they are richly innervated by a highly preserved network of afferent and efferent nerves which con-tribute to valve structure and function[13–15]

Cell lineage and developmental biology Aortic valve originate from endocardial cushions in the devel-oping embryo Endocardial cushions result from the migration

of endothelial cells into the cardiac jelly, followed by a process

of endothelial-to-mesenchymal transformation (EMT) which initiates what will eventually become a tri-layered aortic valve structure Migration, differentiation and delamination of these cells is a tightly controlled series of processes which depend on the activity of specific signaling molecules such as NOTCH1, transforming growth factorb (TGF-b) and the wnt/b-catenin pathway[16–19], as well hemodynamic cues[20] Importantly, the embryonic outflow tract which would ultimately yield the aortic and pulmonary valves as well as the ascending aorta and pulmonary artery are composed of cells derived from the neural crest[21] NOTCH1 is also involved in regulating the migration and differentiation of these cells in the embryo

[17] Understanding the developmental biology of the aortic valve and root has important implications for understanding normal and diseased valve physiology because most of the sig-nals that are operational during morphogenesis continue to influence growth and adaptation in postnatal life[22] For in-stance, EMT continues into adult life as demonstrated by the differentiation of mature VECs into mesenchymal cells expressing smooth musclea-actin, a process which could con-tribute to valve repair and interstitial cell regeneration[23] In addition, defining cell lineages helps explain differences in cell behaviour in response to common stimuli[24]

Aortic valves lie in a unique hemodynamic environment Blood flowing through the vasculature generates shear stress

on the luminal side of the vessel, which is entirely lined by endothelium Shear stress represents the frictional force per unit area The magnitude of shear stress can be estimated in most of the vasculature by Poiseuille’s law stating that wall shear stress is proportional to blood flow viscosity and

volu-Figure 1 Histological section showing the triple layer

architec-ture of a normal aortic valve Endothelial cells form a monolayer

on each side of the cusp Interstitial cells, a mix of fibroblasts,

smooth muscle cells (SMCs) and myofibroblasts fill the body of

the cusp They possess contractile and secretory properties and are

responsible for the synthesis and repair of the surrounding

extracellular matrix

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metric flow rate and inversely proportional to the third power

of the internal radius[25] While actual wall shear stress is very

difficult to estimate, mean shear stress along an artery is

esti-mated at 20 dynes/cm2 Unlike blood vessels, the aortic valve

is not a cylindrical structure, therefore estimation of shear

stress levels on either side of the valve requires more complex

modelling Nevertheless, the pattern of flow on the aortic and

the ventricular sides of the valve has been recognized for a long

time Leonardo da Vinci was the first to accurately describe the

laminar flow on the ventricular surface of the valve and the

dis-turbed vortex flow in the sinuses of Valsalva to which ECs on

the aortic side are exposed Estimates of actual valve shear

stresses have varied significantly in the literature[26–30] Some

have suggested that mean shear stress along the ventricular

surface of the aortic valve is around 20 dynes/cm2

[30,31], while others estimate actual peak shear stress on the

ventricu-lar surface at 80 dynes/cm2whereas it oscillates between +10

and 8 dynes/cm2on the aortic surface [32] Flow along the

ventricular surface is a high-shear laminar flow whereas it is

a low-shear disturbed flow on the aortic side

Endothelial cell heterogeneity

The endothelium is a monolayer of cells lining blood and

lym-phatic vessels which sits at the interface between all body

or-gans and blood They possess thrombogenic,

anti-adhesive, anti-proliferative and vasodilatory properties which

mainly result from the synthesis and release of nitric oxide

(NO), its major biosynthetic product and prostacyclin

(PGI2) Nevertheless, although all endothelial cells throughout

the body share the same basic properties, studies have

demon-strated a considerable amount of heterogeneity between the

endothelium from different regions of the body both at the

structural and functional levels [33] Structurally, although

most endothelial cells have a typical cobblestone appearance,

they can vary significantly in thickness, ranging from 0.1lm

in capillaries and veins to 1lm in the aorta In addition, the

number, distribution and properties of tight and adherens

junctions between them varies significantly from one vascular

bed to another reflecting endothelial adaptation to its

hemody-namic and metabolic environment Junctions are tighter in

large vessels exposed to high-shear stresses and flow rates than

small arterioles, capillaries or venules Finally, the

endothe-lium can be continuous (large arteries and veins) or

discontin-uous (liver sinusoidals), fenestrated or non-fenestrated to allow

filtration and transendothelial transport Similarly, endothelial

cells exhibit marked functional heterogeneity on several levels:

permeability, mechanotransduction pathways in response to

mechanical stimuli and angiogenesis among other functions

Aortic valve endothelial cells exhibit unique properties

Mechanotransduction and alignment

For a long time, aortic VECs were thought to play a minor

role in valve physiology because valves were considered passive

structures that opened and closed in response to changes in

transvalvular pressure More recently, several studies focusing

on the structural and functional properties of aortic VECs

sug-gest that valvular endothelium possesses unique properties

which distinguish it from other endothelial beds, particularly

the endothelium lining the aorta with which it lies in direct

continuity The most striking difference between valvular and vascular endothelial cells is cell alignment with regards

to flow orientation Whereas the vascular endothelium throughout the body aligns with the long axis of the cell par-allel to flow[34](except in areas of turbulent flow)[35], VECs are aligned perpendicular to the direction of flow [31,36]

(Fig 2) This was first described by Deck [36] by electron microscopic analysis of explanted aortic valves and further val-idated by in vitro studies[31] Cultured porcine aortic VECs were compared to aortic (vascular) endothelial cells from the same animal in response to unidirectional non-pulsatile lami-nar flow at 20 dynes/cm2 Whereas vascular cells were aligned parallel to flow after 24 h, instead, VECs aligned perpendicular

to flow even without the presence of an aligned substrate[37] These adaptations were dependent on cytoskeletal reorienta-tion, a process involving different mechanotransduction path-ways in each type of endothelium Laminar flow induced activation of Rho-kinase, phosphatidylinositol-3-kinase and

Figure 2 (a) Scanning electron microscopy of aortic valve endothelial cells forming a monolayer on the surface of the aortic cusps (1000·) The cells are tightly joined and aligned thus constituting the link between the blood milieu and the interstitial space (b) Scanning electron microscopy of aortic valve endothelial cells on the aortic side of the cusp at higher magnification (5000·) Microcilia on the surface of the cells are visible and likely act as sensors of changes in hemodynamic shear stress, in turn activating different intracellular mechanotransduction pathways

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calpain pathways in vascular endothelial cells, whereas VECs

did not require activation of the latter for cytoskeletal

reorga-nization[31]

Gene expression profile

VECs also appear to have a higher proliferation rate than

vas-cular endothelial cells[38] Furthermore, both sets of cells

pos-sess different transcription profiles In a study examining the

transcription profiles of 847 genes, there was common

expres-sion of 55 activated genes whereas a further 48 genes were

dif-ferentially activated Among those genes with a higher

activation in the valvular ECs were transcription factors

associated with higher proliferation rate such as jun D and

protein kinase C [38] Notably, both vascular endothelium

and VECs expressed markers linked with calcification such

as osteonectin, bone morphogenic protein-7 and -9 (BMP-7

and BMP-9) Differences in gene expression profile between

vascular and valvular endothelium were further validated by

another study showing different gene expression profiles in

re-sponse to shear stress stimulation of cultured porcine aortic

endothelial cells or VECs [37] In that study, Butcher et al

showed preferential expression of genes associated with

chon-drogenesis by VECs whereas vascular endothelial cells

ex-pressed more genes associated with osteogenesis Shear stress

reduced the expression of osteogenic genes[37]

Aortic valve endothelial cells are different on both sides of the

valve

Aortic valve calcification is a major clinical problem in elderly

patients and those with bicuspid aortic valve disease With

in-creased life expectancy, improved diagnostic techniques and

better global access to health care, the incidence of aortic valve

calcification is expected to triple within the next 40 years[39],

making it one of the major sources of cardiovascular disease

In recent years, it has become increasingly clear that aortic

valve calcification is an active cell-driven process that shares

many similarities with atherosclerosis Among those, early

endothelial dysfunction has been shown to act as an initial

occurrence in the cascade of events leading to valve

calcifica-tion[40] Histologically, aortic valve calcification occurs

exclu-sively on the aortic side of the valve suggesting that perhaps

VECs on the aortic side are less resistant to calcification than

those on the ventricular side (Fig 3) Simmons et al developed

an innovative technique to separately analyze gene expression

of VECs from either side of the valve[41] This modified

Haut-chen technique for en face isolation of VECs allowed reliable

extraction of high quality mRNA for analysis of gene

expres-sion profiles between both aortic and ventricular VECs The

authors reported the differential expression of 584 genes

in situ between both sides of the valve[42] VECs derived from

the aortic side expressed fewer genes associated with inhibition

of calcification such as oteoprotegerin, parathyroid hormone

and chordin, a protein that inhibits the osteoinductive effects

of BMPs[42] In addition, aortic-sided VECs showed increased

expression of transcripts linked with bone formation including

BMP4 However, this was balanced by a higher expression of

antioxidative and anti-inflammatory genes on the aortic side

Notably, there were higher levels of endothelial nitric oxide

synthase (eNOS) on the aortic side of the valve Because the

mRNA obtained in this study originated directly from freshly

explanted aortic valves, it represented a good picture of in situ gene expression in normal valves However, the gene array used was a human array and only covered 12,000 genes Cur-rently, porcine gene arrays that cover >24,000 genes are com-mercially available and could provide additional clues into the functional side-specificity of aortic VECs This will be greatly enhanced following the full sequencing of the pig genome which is expected towards the end of 2009 We have recently succeeded in separately isolating aortic VECs from either side

of the valve and culturing them in vitro We are currently studying their intrinsic properties in vitro in an effort to deter-mine their responses to various mechanical and pharmacolog-ical stimuli as well as modifications in gene expression profiles Differences in flow patterns between both sides of the valve

as described earlier are sensed by a thin layer of glycoproteins

on the luminal surface of endothelial cells called the glycoca-lyx, which communicates with the cytoskeleton and can activate several signaling pathways in response to flow

[25,43–45] This process of translating mechanical stimuli into biological signals is commonly termed mechanotransduction Studies on vascular endothelial cells have demonstrated that differences in shear stress translate into activation of different signaling pathways, illustrated by the presence of atheroscle-rotic plaques in areas of low wall shear stress in the vasculature such as the carotid artery bifurcation One of the major acti-vated signaling pathways is the nuclear factor-kB (NFkB), a highly conserved transcription factor which translocates to the nucleus when activated, triggering the production of pro-inflammatory molecules[46,47] To date, the glycocalyx has not been identified or characterized on the surface of aortic valves Identification of the specific cell-surface molecules which contribute to mechanotransduction in VECs could open new avenues into understanding disease processes in condi-tions of abnormal shear stresses on the valves

The different mechanotransduction pathways involved on either side of the valve in response to different patterns of flow have yet to be described (Fig 4) Recently, a flow apparatus consisting of a cone and plate was developed to allow

in vitro reproduction of aortic and ventricular flow patterns

Figure 3 Histological section of a calcified human aortic valve from a patient with calcified aortic valve stenosis Tripp–McKay silver impregnation staining shows the calcium nodule on the aortic side of the cusp, below the endothelial layer

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on aortic valves [32] Preliminary studies using that system

show that unlike vascular endothelium, VECs on both the

aor-tic and ventricular side were not activated by exposure to

bidi-rectional oscillatory flow (reproducing flow on the aortic side),

as illustrated by an absence of vascular adhesion molecule-1

(VCAM-1), intercellular adhesion molecule-1 (ICAM-1),

E-selectin and BMP4[48] However, exposure of aortic-sided

endothelium to high-shear laminar flow (ventricular flow)

resulted in endothelial activation characterized by expression

of these inflammatory markers[48] This underscores the

no-tion that both sets of VECs activate different

mechanotrans-duction pathways in response to similar mechanical stimuli

Endothelial cells play an important role in normal aortic valve

function

In addition to their traditional role in preserving an

anti-adhe-sive and anti-proliferative surface, VECs are actively involved

in the regulation of aortic valve function As discussed

previ-ously, mechanical strain on the surface of the cusps as well

as coronary blood flow are both affected by the shape and

stiffness of the aortic valve cusps[5,6,49,50] Aortic valve

cal-cification has been shown to occur in areas of high mechanical

strains on the valve, namely the commissures, the base of the

cusps and the free edges[51] Changes in the stiffness of the

aortic valve cusps could significantly impact stress magnitude

and distribution along the surface of the cusps We have

recently demonstrated the role of the endothelium in

regulat-ing the mechanical properties of the aortic valve in vitro usregulat-ing

a biaxial micromechanical testing system [52] In that study,

aortic valve cusps were stimulated with endothelin-1 (ET-1),

a potent vasoconstrictor peptide which is released by

endothe-lial cell or serotonin (5-HT), an agent which mediates the re-lease of nitric oxide by the endothelium Addition of ET-1 resulted in a 25% increase in the elastic modulus (stiffness)

of aortic valve cusps, whereas addition of 5-HT induced a 30% decrease in valve stiffness Interestingly, addition of cyto-chalasin D, an inhibitor of actin polymerization reversed the increase in valve stiffness in response to ET-1, highlighting the role of the contractile elements in the VIC population and the communication between VECs and VICs in the valve These findings suggest that the aortic valve is capable of auto-regulation, but that it is also subject to overall systemic conditions such as hypertension or diabetes which are often accompanied by dysregulated concentrations of circulating vasoactive agents As previously stated, hypertension, smoking and diabetes are among the risk factors associated with aortic valve calcification In addition to their direct metabolic effects, this could be secondary to their effect in modulating valve stiff-ness which could lead to abnormal stress distribution along the cusps[53]

Is the aortic valve endothelium involved in valve calcification?

To date, no definitive link between VECs and valve calcifica-tion has been directly established Nevertheless, a number of studies have recently demonstrated a potential causal relation-ship Taken together, this cumulative weight of evidence strongly suggests that the endothelium plays an active role in the cellular and molecular events involved in aortic valve cal-cification As mentioned earlier, Simmons et al showed a dif-ferent pattern of gene expression between VECs on either side

of the valve characterized by a higher expression of pro-calcific genes and on the aortic side of the valve[42] In addition, an

Figure 4 Illustration of some of the mechanostransduction pathways activated inside VECs in response to shear stress Cells sense shear stress through mechanosensors on their surface which trigger activation of intracellular mediators such as nuclear factor-kB (NFkB) or Rho, which lead to synthesis of various signaling molecules which are either directly released by the cells or translocate to the nucleus and induce modifications in gene expression The pathways activated result in different effects including modified chemotaxis, cell adhesion and expression of inflammatory markers (adapted from[46,62])

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in vitro study examining the role of strain on VEC activation

in vitro showed that overstretching of the cells induces the

expression of adhesion molecules, an important initial event

leading to local inflammation which is characterized by

mono-cyte chemotaxis and infiltration [54] Physiological strain

which was evaluated as a cyclical stretch of 10% did not affect

VECs, whereas 20% cyclical strain on resulted in expression of

VCAM-1, ICAM-1 and E-selectin[54], similar to what is

com-monly observed in calcified human valves In addition, a recent

study by Kennedy et al using an in vitro model of VIC

calci-fication such as described by us and others [55,56],

demon-strated that addition of nitric oxide donors or agents raising

intracellular cyclic guanosine monophosphate (cGMP) levels

led to a decrease in the formation of calcified nodules in

re-sponse to stimulation with osteogenic medium and

transform-ing growth factorb (TGFb), an important pathogenic element

in vascular and valvular calcification[57] Furthermore, in our

laboratory, we developed an in vitro model of aortic valve

cal-cification by exposing whole cusp tissue to osteogenic medium

for a period of 10–14 days Preliminary data show that similar

to what is observed in vivo, calcium nodules (characterized by

positive Alizarin-red staining) were present on the aortic side

of the valve More importantly, endothelial denudation of

the cusp surface using a cell scraper or addition of L-NAME

(an inhibitor of nitric oxide synthase) led to a significant

in-crease in the number of Alizarin-red positive nodules (data

not published)

Taken together, these results strongly suggest that a healthy

and functional valve endothelium acts an important first-line

barrier against valve calcification The endothelium appears

to exert its effect by a combination of effects on overall valve

structure and function It can modify the mechanical

proper-ties of the cusps which might have an important effect in

pro-tecting the leaflets from high mechanical stresses These

elevated stresses on the cusps can eventually lead to areas of

‘‘micro-tears’’ or interruption of the endothelial barrier As a

consequence, the endothelium which appears to protect

against calcification by communicating with the VICs in a

par-acrine way loses this capacity and exposes the VICs to

osteo-genic stimuli VICs have been shown to respond to direct

osteogenic stimuli by expressing a high number of activated

fibroblasts, the myofibroblasts These cells have the capacity

to transdifferentiate into osteogenic-like cells and produce

hydroxyapatite

Clinical implications

The importance of the living cellular environment in the valve

is aptly illustrated in patients requiring aortic valve

replace-ment surgery[58] Most aortic valve prostheses are acellular,

including tissue valve prostheses and even aortic homografts

which become rapidly decellularized These different valve

sub-stitutes are hampered by their limited durability due to

struc-tural valve deterioration [59] However, one operation

consists of replacing the diseased aortic root with the patient’s

pulmonary root: the Ross procedure It is the only operation

which guarantees long-term viability of the neo-aortic valve

This in turn directly translates into a significant benefit to

the patients in terms of survival, freedom from reoperation,

freedom from valve-related complications and quality of life

[60] We believe that this observed benefit is in large part due

to the ability of the neo-aortic root to adapt to its hemody-namic environment, to respond to various stimuli and to con-tinuously repair in a similar fashion to normal valves[61] Nevertheless, the Ross procedure is a technically complex operation which results in uneven results among different centers around the world Therefore, a more reproducible approach is necessary This will be best addressed by progresses

in heart valve tissue engineering Although some promising first steps were made, more is required both in understanding normal valve physiology and combining biological, biochemi-cal, engineering, physics and nanotechnology expertise to develop a biocompatible tri-layered aortic valve substitute that can reproduce the sophisticated functions of the normal aortic valve We and others are making big strides towards reaching that goal and are optimistic that a fully tissue-engineered heart valve will eventually become a viable option for patients undergoing valve replacement surgery

Finally, therapeutic strategies targeting the endothelium could have a major impact on the treatment of aortic valve dis-ease Various agents have shown promise in reducing, revers-ing or slowrevers-ing the progression of vascular atherosclerotic disease, including statins[55] In addition to their lipid-lower-ing effect, statins exhibit a range of pleiotropic effects which can have a direct impact on endothelial function and address the different pathological mechanisms associated with hyper-cholesterolemia which lead to valve calcification, such as apop-tosis [62] Thus far, most clinical trials have shown mixed results for the use of statins in patients with aortic valve dis-ease However, it is possible that statin administration needs

to be initiated as primary prevention in order to show a reduc-tion in the incidence of aortic valve disease In the mean time, search for other more endothelial-specific compounds should

be undertaken and will only be possible following a thorough understanding of the specific biology of VECs

Conclusion The aortic valve lies at a critical junction in the circulatory sys-tem and is subjected to extremes of mechanical and hemody-namic forces with every cardiac cycle Yet, in the majority of people, these thin structures never fail This is a result of the highly sophisticated cellular and molecular functions of aortic valves We have overviewed the contribution of the endothe-lium to the structure and function of aortic valves in health and disease As our knowledge of these cells both in vitro and in vivo increases, understanding of the pathophysiological mechanisms of aortic valve disease will become clearer It is hoped that this will open opportunities towards targeted ther-apeutic approaches to aortic valve disease, starting with tissue engineering of heart valves

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