Growth-related micromorphological characteristics of the porcine common carotid artery.

The purpose of this study was to gain knowledge about the micromorphology of the porcine common carotid artery (CCA) during the period of growth over the bodyweight range of 10-40 kg. CCA samples from German landrace pigs (DL) aged either 2 or 3 months (DL-2 and DL-3) were compared with samples from Göttingen minipigs (GM) aged either 18 or 40 months (GM-18 and GM-40) using transmitted light (phase-contrast mode) and transmission electron microscopy. The GM-18, GM-40 and the DL-3 groups had typical muscular artery histological characteristics. Contrasting to this, the 2-month-old DL pigs had a transitional artery type being characterized by a significantly higher proportion of elastic fibres and a significantly lower number of smooth muscle cells than did the 1 month older DL-3. During the period of maturation, the tunica media of the CCA in GM animals thickened by 1.3× and in DL animals by 2.5× resulting in an overall increased vessel wall thickness. The cumulated thickness of the tunica interna (endothelium, stratum subendotheliale and internal elastic lamina) and the tunica media (including the external elastic lamina) of DL-3 and GM-40 pigs were similar to each other and comparable to that of humans. With an increasing vessel wall thickness, the luminal diameter decreased in GM by 19% and in DL by 11%. Additionally, in the older age groups, GM-40 and DL-3, the internal elastic lamina principally was continuous, but there were also interrupted large segments of elastic lamina separated by gaps. In addition, the principal internal elastic lamina was duplicated in several places.

Knockout of Density-Enhanced Phosphatase-1 impairs cerebrovascular reserve capacity in an arteriogenesis model in mice.

Collateral growth, arteriogenesis, represents a proliferative mechanism involving endothelial cells, smooth muscle cells, and monocytes/macrophages. Here we investigated the role of Density-Enhanced Phosphatase-1 (DEP-1) in arteriogenesis in vivo, a protein-tyrosine-phosphatase that has controversially been discussed with regard to vascular cell biology. Wild-type C57BL/6 mice subjected to permanent left common carotid artery occlusion (CCAO) developed a significant diameter increase in distinct arteries of the circle of Willis, especially in the anterior cerebral artery. Analyzing the impact of loss of DEP-1 function, induction of collateralization was quantified after CCAO and hindlimb femoral artery ligation comparing wild-type and DEP-1(-/-) mice. Both cerebral collateralization assessed by latex perfusion and peripheral vessel growth in the femoral artery determined by microsphere perfusion and micro-CT analysis were not altered in DEP-1(-/-) compared to wild-type mice. Cerebrovascular reserve capacity, however, was significantly impaired in DEP-1(-/-) mice. Cerebrovascular transcriptional analysis of proarteriogenic growth factors and receptors showed specifically reduced transcripts of PDGF-B. SiRNA knockdown of DEP-1 in endothelial cells in vitro also resulted in significant PDGF-B downregulation, providing further evidence for DEP-1 in PDGF-B gene regulation. In summary, our data support the notion of DEP-1 as positive functional regulator in vascular cerebral arteriogenesis, involving differential PDGF-B gene expression.

External aortic root support: a histological and mechanical study in sheep.

OBJECTIVES: Personalized external aortic root support has completed initial evaluation and has technology appraisal in the UK for patients with Marfan syndrome for use as an alternative to root replacement. Its long-term success in preventing aortic dissection remains uncertain. Here, we report a study in sheep to establish whether the externally supporting mesh, as used clinically, is biologically incorporated. The strength of the resulting mesh/artery composite has been tested. METHODS: The carotid artery of growing sheep (n=6) was enclosed in a mesh sleeve made of a polymer, polyethylene terephthalate. After a predefined interval of 4-6 months, a length of the artery was excised, including the sleeved and unsleeved portions, and was stress tested and examined histologically. RESULTS: One animal died of pneumonia 7 days after implantation. Comparing sleeved with normal segments, the overall thickness was increased and there was a fibrotic sheet in the periarterial space. The overall vessel wall architecture was preserved in all specimens. Although media thickness of ensleeved arteries was smaller and in one animal mild oedema was found in one quadrant of the outer part of the media. There was a significant increase in stiffness and maximum tensile strength of the supported segments compared with normal arterial tissue. CONCLUSIONS: Polyethylene terephthalate mesh, as used for the external support of the dilated aortic root in Marfan syndrome, becomes incorporated in the periadventitial tissue of the carotid artery of sheep. Limited thinning of the media, without any signs of inflammation or medial necrosis, was visible. There was a significantly greater tensile strength in the carotid artery/mesh composite compared with the unsleeved carotid artery.

Time course of carotid artery growth and remodeling in response to altered pulsatility.

Elucidating early time courses of biomechanical responses by arteries to altered mechanical stimuli is paramount to understanding and eventually predicting long-term adaptations. In a previous study, we reported marked long-term (at 35-56 days) consequences of increased pulsatile hemodynamics on arterial structure and mechanics. Motivated by those findings, we focus herein on arterial responses over shorter periods (at 7, 10, and 14 days) following placement of a constrictive band on the aortic arch between the innominate and left carotid arteries of wild-type mice, which significantly increases pulsatility in the right carotid artery. We quantified hemodynamics in vivo using noninvasive ultrasound and measured wall properties and composition in vitro using biaxial mechanical testing and standard (immuno)histology. Compared with both baseline carotid arteries and left carotids after banding, right carotids after banding experienced a significant increase in both pulse pressure, which peaked at day 7, and a pulsatility index for velocity, which continued to rise over the 42-day study despite a transient increase in mean flow that peaked at day 7. Wall thickness and inner diameter also increased significantly in the right carotids, both peaking at day 14, with an associated marked early reduction in the in vivo axial stretch and a persistent decrease in smooth muscle contractility. Glycosaminoglycan content also increased within the wall, peaking at day 14, whereas increases in monocyte chemoattractant protein-1 activity and the collagen-to-elastin ratio continued to rise. These findings confirm that pulsatility is an important modulator of wall geometry, structure, and properties but reveal different early time courses for different microscopic and macroscopic metrics, presumably due to the separate degrees of influence of pressure and flow.

The relationship between the internal jugular vein and common carotid artery in the carotid sheath: the effects of age, gender and side.

The internal jugular vein is often used for central venous catheter placement. The variations in the location of this vein along the major neck vessels (in the carotid sheath) may account for unsuccessful cannulations or iatrogenic arterial injuries. The aim of this study was to delineate the relation of the internal jugular vein and common carotid artery in the lower neck, and to assess the effects of age, gender and side on these anatomical structures. Two-dimensional ultrasonographic examinations of the right and left supraclavicular triangle were performed in 219 adult individuals who had no history of neck surgery or known pathology. The location of the internal jugular vein in relation to the common carotid artery was recorded. An anterolateral location of the internal jugular vein was the most common configuration observed on both sides (84% right side and 91.8% left side) followed by the lateral (14.2% right and 6.4% left) and anterior (1.4% right and 1.8% left) locations. A single case of a medial internal jugular vein was observed on the right side (0.23% of both sides). Subjects with a laterally located internal jugular vein were older than those with an anterolateral configuration (P<0.01). No gender differences were found with regard to these two configurations (P=0.867). The laterally located internal jugular vein was more frequent on right sides (P=0.007). Such information may be potentially useful for clinicians who are managing critically ill patients or patients undergoing hemodialysis.

Effects of elastin haploinsufficiency on the mechanical behavior of mouse arteries.

Supravalvular aortic stenosis (SVAS) is associated with decreased elastin and altered arterial mechanics. Mice with a single deletion in the elastin gene (ELN(+/-)) are models for SVAS. Previous studies have shown that elastin haploinsufficiency in these mice causes hypertension, decreased arterial compliance, and changes in arterial wall structure. Despite these differences, ELN(+/-) mice have a normal life span, suggesting that the arteries remodel and adapt to the decreased amount of elastin. To test this hypothesis, we performed in vitro mechanical tests on abdominal aorta, ascending aorta, and left common carotid artery from ELN(+/-) and wild-type (C57BL/6J) mice. We compared the circumferential and longitudinal stress-stretch relationships and residual strains. The circumferential stress-stretch relationship is similar between genotypes and changes <3% with longitudinal stretch at lengths within 10% of the in vivo value. At mean arterial pressure, the circumferential stress in the ascending aorta is higher in ELN(+/-) than in wild type. Although arterial pressures are higher, the increased number of elastic lamellae in ELN(+/-) arteries results in similar tension/lamellae compared with wild type. The longitudinal stress-stretch relationship is similar between genotypes for most arteries. Compared with wild type, the in vivo longitudinal stretch is lower in ELN(+/-) abdominal and carotid arteries and the circumferential residual strain is higher in ELN(+/-) ascending aorta. The increased circumferential residual strain brings the transmural strain distribution in ELN(+/-) ascending aorta close to wild-type values. The mechanical behavior of ELN(+/-) arteries is likely due to the reduced elastin content combined with adaptive remodeling during vascular development.

A mixture model of arterial growth and remodeling in hypertension: altered muscle tone and tissue turnover.

Hypertension results in a thickening of the arterial wall due to a net increase in wall constituents via a coordinated production and removal of smooth muscle and extracellular matrix. Although many reports address the associated changes in material properties, few models address the biomechanics of the growth and remodeling process. In this paper, we employ a new, fundamentally different approach to modeling arterial adaptation in hypertension. In particular, basic, characteristic features of hypertension are simulated using a constrained mixture model wherein individual constituents can turnover at different rates and can have different natural configurations. We show, for example, that if there is no turnover of elastin (which approximates responses in maturity), the model vessel is able to reduce an early elevation in wall stress via a thickening of the wall even though the adaptation may be suboptimal. Conversely, if all constituents can turnover completely (which may approximate late development), the model vessel can restore the state of stress and material properties to native values. Given the potential of such a model, there is a need for more data on the history of turnover of individual constituents and their individual material properties.

Effects of maturation on mechanisms of cGMP-induced cerebral vasodilatation.

In light of observations that cerebrovascular levels of cGMP vary during maturation, the present study examines the possibility that the mechanisms mediating cGMP-induced cerebral vasodilatation also change during maturation. Specifically, these experiments explore age-related changes in the ability of cGMP to both: (1) depress cytosolic calcium concentration, and (2) attenuate contractile protein calcium sensitivity in alpha-toxin and beta-escin permeabilized preparations as well as fura-2 loaded arteries. The present data demonstrate that: (1) cGMP attenuates cytosolic calcium concentration at lower concentrations than required to reduce myofilament calcium sensitivity; (2) both potassium-induced and 5HT-induced contractions were more sensitive to cGMP in fetal than adult arteries; (3) all potassium-induced increases in cytosolic calcium were resistant to the effects of cGMP, but those produced by 5HT were sensitive to attenuation by cGMP, and more so in fetal than in adult basilar arteries, and (4) cGMP attenuated both basal and agonist-enhanced myofilament calcium sensitivity. Overall, these data demonstrate that the mechanisms mediating the multiple vasoactive effects of cGMP are more potent in immature than in mature cerebral arteries and are heavily influenced by both the artery type and the method of contraction.

Electron microscopic study on the development of the carotid body and glomus cell groups distributed in the wall of the common carotid artery and its branches in the chicken.

Development of the carotid body and the glomus cell groups in the wall of the common carotid artery and its branches was studied in chickens at various developmental stages by electron microscopy. At 8 days of incubation, the carotid body anlage consisted of mesenchyme-like cells, whereas the clusters of epithelial cells, which occasionally contained a few dense-cored vesicles and were accompanied by unmyelinated nerve fibers, were located in the region surrounding the carotid body anlage and in the wall of the common carotid artery. Subsequently, the granule-containing cells together with nerve fibers were detected in the periphery of the carotid body anlage. At 12 days of incubation, a large number of granule-containing cells (glomus cells) were dispersed throughout the carotid body parenchyma and were also widely distributed along the common carotid artery and its branches. The cells frequently extended long cytoplasmic processes that made contact with other glomus cells and nerve fibers. Synaptic junctions which showed desmosome-like thickening of pre- and postsynaptic membranes and accumulations of small clear vesicles (around 50 nm in diameter) were first detected along the contact between the long axons and glomus cells at 12 days of incubation. In the wall of the common carotid artery, interdigitations between the cytoplasmic processes of glomus cells and smooth muscle cells began to form. Sustentacular cells investing partly the glomus cells were also discerned both in the carotid body and around the arteries at this stage. At 14 days of incubation, the glomus cells expressed most of the characteristics of the mature cells, and the synaptic junctions displaying afferent morphology appeared; the secretory granules of glomus cells were accumulated near and attached to the desmosome-like thickening of apposed membranes.

Effects of maturation on cyclic GMP-dependent vasodilation in ovine basilar and carotid arteries.

The present experiments examine the effects of maturation on cyclic GMP (cGMP)-mediated vasodilation in 688 segments of common carotid (COM) and basilar (BAS) arteries taken from newborn (3- to 7-d-old) and nonpregnant adult sheep. The main finding is that maximum efficacy for relaxation decreased with maturation in both artery types for the nitric oxide releasing vasodilators S-nitroso-N-acetyl-penicillamine and nitroglycerin. These decreases could not be explained by changes in the -log ED50 concentrations for either vasodilator. Determination of the time course of cGMP responses to S-nitroso-N-acetyl-penicillamine or nitroglycerin at 10 microM revealed that the peak cGMP responses to these agents (range: 5.3 +/- 0.8 to 8.3 +/- 1.6 pmol/mg of protein) also did not vary significantly with age. However, cGMP attained peak values more rapidly in adult (COM: 50 s; BAS 30 s) than in newborn (COM: 60-80 s: BAS, 40-60 s) segments and returned to baseline more slowly in newborn than in adult segments, suggesting that maturation accelerates cGMP turnover. Correspondingly, baseline levels of cGMP were higher in newborn (COM: 1.0 +/- 0.1; BAS: 3.3 +/- 0.5 pmol/mg of protein) than in adult (COM: 0.3 +/- 0.1; BAS: 1.7 +/- 0.2 pmol/mg of protein) segments. Despite these differences in cGMP time course, rates of relaxation in response to S-nitroso-N-acetyl-penicillamine and nitroglycerin did not vary significantly with age, indicating that the temporal relation between cGMP and relaxation is different in newborn and adult arteries.(ABSTRACT TRUNCATED AT 250 WORDS)