Density Of Elastic Fibers Research Articles

Systemic sclerosis and idiopathic interstitial pneumonia: histomorphometric differences in lung biopsies

The aim of this study was to examine the parenchymal and extracellular matrix remodeling process in two histologic patterns-nonspecific interstitial pneumonia (NSIP) and usual interstitial pneumonia (UIP)-in cases of idiopathic and sclerosis/systemic sclerosis (SSc)-associated interstitial pneumonia. We examined 15 cases of idiopathic NSIP, 10 cases of idiopathic UIP, 5 cases of SSc-UIP and 9 cases of SSc-NSIP. In the lung parenchyma, epithelial cells, endothelial cells and myofibroblasts were evaluated by immunohistochemical staining, whereas histochemical staining was used in order to evaluate collagen/elastic fibers in the extracellular matrix. The percentage of surfactant protein A-positive epithelial cells was significantly greater in idiopathic NSIP than in SSc-NSIP, as well as being greater in idiopathic UIP than in SSc-UIP. Idiopathic NSIP and idiopathic UIP presented significantly higher immunoexpression of alpha smooth muscle actin in myofibroblasts than did SSc-NSIP and SSc-UIP. The percentage of CD34 endothelial cells in the pulmonary microvasculature was significant lower in idiopathic UIP than in SSc-UIP. The density of collagen fibers was significantly greater in idiopathic NSIP and idiopathic UIP than in SSc-NSIP and UIP. In contrast, the elastic fiber density was significantly lower in idiopathic UIP than in SSc-UIP. Increased collagen synthesis, destruction of elastic fibers, high myofibroblast proliferation and poor microvascularization might represent a remodeling process found in idiopathic interstitial pneumonia, whereas the reverse might represent a repair process in SSc-associated interstitial pneumonia.

Nanoindentation of histological specimens: Mapping the elastic properties of soft tissues

Although alterations in the gross mechanical properties of dynamic and compliant tissues have a major impact on human health and morbidity, there are no well-established techniques to characterize the micromechanical properties of tissues such as blood vessels and lungs. We have used nanoindentation to spatially map the micromechanical properties of 5-mum-thick sections of ferret aorta and vena cava and to relate these mechanical properties to the histological distribution of fluorescent elastic fibers. To decouple the effect of the glass substrate on our analysis of the nanoindentation data, we have used the extended Oliver and Pharr method. The elastic modulus of the aorta decreased progressively from 35 MPa in the adventitial (outermost) layer to 8 MPa at the intimal (innermost) layer. In contrast, the vena cava was relatively stiff, with an elastic modulus >30 MPa in both the extracellular matrix-rich adventitial and intimal regions of the vessel. The central, highly cellularized, medial layer of the vena cava, however, had an invariant elastic modulus of ~20 MPa. In extracellular matrix-rich regions of the tissue, the elastic modulus, as determined by nanoindentation, was inversely correlated with elastic fiber density. Thus, we show it is possible to distinguish and spatially resolve differences in the micromechanical properties of large arteries and veins, which are related to the tissue microstructure.

Estudo imunohistoquímico do remodelamento pulmonar em camundongos expostos à fumaça de cigarro

Various studies of emphysema involve long-term exposure of animals to cigarette smoke, focusing on the cell type involved in the protease/antiprotease imbalance and on extracellular matrix degradation. In emphysema, increased expression of metalloproteinases is associated with cytokines, and evidence suggests that the matrix metalloproteinase-12 (MMP-12) plays an important role. Our objective was to investigate tissue inhibitor of metalloproteinase-2 (TIMP-2), tumor necrosis factor-alpha (TNF-alpha) and interleukin-6 (IL-6) detection by immunohistochemical methods in mouse lung. Male C57BL/6 mice were exposed 3 times a day to smoke of 3 cigarettes over a period of 10, 20, 30 or 60 days in an inhalation chamber (groups CS10, CS20, CS30 and CS60, respectively). Controls were exposed to the same conditions in room air. A progressive increase in the number of alveolar macrophages was observed in the bronchoalveolar lavage fluid of the exposed mice. The mean linear intercept, an indicator of alveolar destruction, was greater in all exposed groups when compared to control group. In the CS10, CS20 and CS30 mice, the immunohistochemical index (II) for MMP-12 increased in parallel with a decrease in II for TIMP-2 in the CS10, CS20 and CS30 mice. The II for the cytokines TNF-alpha and IL-6 was greater in all exposed groups than in the control group. Emphysema, with changes in volume density of collagen and elastic fibers, was observed in the CS60 group. These findings suggest that cigarette smoke induces emphysema with major participation of TNF-alpha and IL-6 without participation of neutrophils.

Nanoindentation of Histological Specimens using an Extension of the Oliver and Pharr Method

AbstractThe micro-mechanical properties of 5 μm thick histological sections of ferret aorta and vena cava were mapped as a function of distance from the outer adventitial layer using nanoindentation. In order to decouple the effect of the glass substrate on the elastic modulus of these thin sections, the nanoindentation data were analyzed using the extended Oliver and Pharr method which is readily accessible for coatings and layered materials with the software package, FilmDoctor®. In the aorta, the elastic modulus was found to decrease progressively from 35 MPa at the adventitia (outermost layer) to 8 MPa at the intima (innermost layer). This decrease in modulus was inversely correlated with elastic fibre density. In contrast, in the vena cava, the stiffest regions were found to be the adventitial (outer) and intimal (innermost) sections of the vessel cross-section. Both these regions were enriched in ECM components. The central region, thought to be largely cellular, had a relatively constant modulus of around 20 MPa. This study demonstrates that with this methodology it is possible to distinguish micro-mechanically between large arteries and veins, and therefore the same approach should allow age or disease related changes in the mechanical properties within a tissue to be quantified.

Mechanical ventilation uncouples synthesis and assembly of elastin and increases apoptosis in lungs of newborn mice.

Prolonged mechanical ventilation (MV) with O2-rich gas inhibits lung growth and causes excess, disordered accumulation of lung elastin in preterm infants, often resulting in chronic lung disease (CLD). Using newborn mice, in which alveolarization occurs postnatally, we designed studies to determine how MV with either 40% O2 or air might lead to dysregulated elastin production and impaired lung septation. MV of newborn mice for 8 h with either 40% O2 or air increased lung mRNA for tropoelastin and lysyl oxidase, relative to unventilated controls, without increasing lung expression of genes that regulate elastic fiber assembly (lysyl oxidase-like-1, fibrillin-1, fibrillin-2, fibulin-5, emilin-1). Serine elastase activity in lung increased fourfold after MV with 40% O2, but not with air. We then extended MV with 40% O2 to 24 h and found that lung content of tropoelastin protein doubled, whereas lung content of elastin assembly proteins did not change (lysyl oxidases, fibrillins) or decreased (fibulin-5, emilin-1). Quantitative image analysis of lung sections showed that elastic fiber density increased by 50% after MV for 24 h, with elastin distributed throughout the walls of air spaces, rather than at septal tips, as in control lungs. Dysregulation of elastin was associated with a threefold increase in lung cell apoptosis (TUNEL and caspase-3 assays), which might account for the increased air space size previously reported in this model. Our findings of increased elastin synthesis, coupled with increased elastase activity and reduced lung abundance of proteins that regulate elastic fiber assembly, could explain altered lung elastin deposition, increased apoptosis, and defective septation, as observed in CLD.

Intraluminal plugs in idiopathic and secondary organizing pneumonia: repair or remodelling?

Organizing pneumonia (OP) is an inflammatory lung disease characterized histologically by intraluminal plugs involving alveolar ducts and alveoli. The aim was to examine extracellular matrix repair and remodelling in 12 cases of idiopathic OP and compare these with 11 cases of secondary OP. Collagen/elastic fibre density, myofibroblast proliferation, microvascular density and endothelial activity in the intraluminal plugs were evaluated by histochemistry, immunohistochemistry and morphometry. The density of the collagen system fibres was greater in plugs of idiopathic OP when compared with secondary OP (P < 0.001). Quantification of myofibroblastic cells confirmed differences observed in patterns of immunoexpression; when compared with secondary OP, idiopathic OP contained fewer myofibroblastic cells in intraluminal plugs (P = 0.01). Microvascular density (CD34) and endothelial activity (vascular cell adhesion molecule-1 and E-selectin) were significantly greater in secondary OP (P < 0.05). Idiopathic and secondary OP show significant variation in morphological features that may represent different responses to injury. Increased collagen synthesis, low myofibroblast proliferation, poor microvascularization and minimal endothelial activity found in idiopathic OP may be more of a remodelling process than secondary OP repair.

Structural analysis of gubernaculum testis in cryptorchid patients submitted to treatment with human chorionic gonadotrophin

To make a histological and stereological analysis of gubernaculum testis elastic system fibers, collagen and striated muscle cells in patients with cryptorchidism treated with human chorionic gonadotrophin (hCG). Gubernacula tissue samples were obtained from 12 patients with cryptorchidism. Patients' ages ranged from 1 to 3 years (mean 1.8). Of the 12 patients, 6 were treated with hCG. The samples were stained with Masson's trichrome to highlight muscle fibers and collagen, and Weigert's resorcin-fuchsin to highlight the elastic system fibers. The volumetric density of these components was determined by stereological methods. The volumetric density of collagen was increased in patients treated with hCG, ranging from 85.62% to 94.48%, while in patients not submitted to hCG treatment the volume density ranged from 52.12% to 89.46% (p=0.0052). The volumetric density of the elastic system fibers was higher in patients treated with hCG, ranging from 9.62% to 19.62%, while patients not submitted to hCG treatment the volume density of elastic system fibers was between 10.38% and 12.38% (p=0.0030). The volume density of striated muscle fibers in patients treated with hCG ranged from 4.76% to 39.16%, while and in patients not treated hCG it ranged from 3.24% to 11.14% (p=0.0052). Gubernacular components alter significantly when submitted to treatment with hCG. Patients who underwent hCG treatment and had no complete testicular migration had an increase in the concentration of elastic and striated muscle fibers and a decrease in the volumetric density of collagen.

Staining Behavior and Distribution of Elastic Fibers in the Pig Skin Dermis

For histological demonstration of elastic fibers in the skin, different investigators use a variety of staining procedures. However, some of these popular methods have pro­duced equivocal results for staining sections of mammalian and human skin. The current study concerned the elastic fiber distribution in the skin of the domestic pig from specimens fixed in four different histological fixatives and subjected to paraffin microtomy at various thickness levels and staining by three commonly used procedures, i.e., aldehyde fuchsin, orcein, and Verhoeff elastic stain. An interconnected elastic fiber network was demonstrated in thick paraffin sections (25–60 μm), whereas thin sections (5–7 μm) presented a disrupted or discontinuous fiber profile. All the staining procedures produced satisfactory results on thin sections. For thicker sections, the most satisfactory results were obtained with aldehyde fuchsin procedure after preoxidation. Preliminary manual morphometric measurement of elastic fiber density with an eyepiece reticle revealed a significantly greater quantity in thicker sections. The distri­bution pattern of elastic fibers in this animal model seems to simulate that reported for human skin. For future experimental procedures with pigskin to explore skin expansion, or dermabrasion, thick paraffin sections stained with aldehyde fuchsin method will reveal more meaningful information on elastic fiber distribution and better quantitative es­timation

Remodeling of the human dermis after application of salicylate silanol

Recently, a controlled double-blind study in patients with photo-aged facial skin demonstrated the beneficial role of oral intake of silanol for skin, hair and nails. The aim of our pilot study was to investigate histologic alterations in human skin after injection of silanol. Seven healthy female caucasian volunteers with a moderate degree of photoaged skin received ten sessions of weekly injections of 0.1% salicylate silanol in the left ventral lateral forearm. The histologic features of punch biopsies of the treated area and the nontreated contralateral arm were compared and the collagen and elastic fibers quantified. Texture analysis was performed on digitalized microscopic images by analyzing the Sarkar fractal dimension or amplitudes (inertia values) after Fast Fourier transformation. The treated area revealed a statistically significant increase of the density of both collagen and elastic fibers. Texture analysis showed more compact and homogeneously distributed collagen fibers after silicon injection. Our results suggest that the application of silicon may stimulate the production of collagen and elastic fibers leading to remodeling of the dermal fiber architecture, which may explain the improvement of the skin surface observed in clinical studies.

Estudo histomorfométrico seqüencial da artéria torácica interna esquerda

OBJETIVO: O uso de enxertos de artéria torácica interna esquerda (ATIE) é atualmente reconhecido como a melhor opção na cirurgia de revascularização miocárdica, proporcionando menor incidência de eventos cardiovasculares e maior sobrevida. Entretanto, com a expansão do uso dos enxertos arteriais, faz se necessário melhor entendimento da estrutura histológica da ATIE. Portanto, o objetivo deste trabalho foi estudar a estrutura histológica da ATIE e a análise histomorfométrica seqüencial e comparativa entre os diferentes segmentos. MÉTODO: Foram estudados espécimes de ATIE retirados de 18 cadáveres, divididos em nove segmentos proporcionais. Cortes de cada segmento foram corados com técnica de Verhoeff-Van Gieson. Foram analisados os seguintes parâmetros: perímetro da luz arterial, espessura da íntima, espessura da camada média, número de fibras elásticas da camada média e a densidade de fibras elásticas. RESULTADOS: Os dados obtidos mostraram que perímetro da ATIE mostra tendência a diminuir ao longo de sua extensão, a espessura da íntima é aumentada nos segmentos proximais, a espessura da camada média diminui ao longo de sua extensão, o número de lâminas elásticas se concentra nos segmentos intermediários, a densidade de lâminas elásticas é diminuída nos segmentos proximais e distais. CONCLUSÃO: Portanto, os achados do presente estudo confirmam a heterogeneidade da estrutura histológica seqüencial da ATIE.

Quantitative analysis of the elastic fibres in the human temporomandibular articular disc and its attachments

A quantitative study of the elastic fibres found in the human temporomandibular disc and its attachments was performed. Seven left discs from 57- to 82-year-old subjects, without macroscopic evidence of a TMJ disorder, were analysed and prepared in parasagittal sections. The surface amount was measured, thresholded and expressed from 0 to 1, using microscopic digitized views after Weigert's resorcin–fuchsin staining of elastic fibres. Fibre density rates ranged from 0 to 0.687. The mean density was 0.1532 ( σ = 0.1150) in the upper bilaminary zone, 0.1097 ( σ = 0.1159) in the lower bilaminary zone, 0.0474 ( σ = 0.0782) in the anterior band, 0.0180 ( σ = 0.0603) in the posterior band and null in the intermediate zone. The difference in density rate between the structures was significant, except for the posterior band and the intermediate zone. The elastic fibre density rates in central and medial locations of the upper and lower bilaminary zones were twice as big as in the lateral locations. In the anterior band, the elastic fibre density was less abundant medially than in its lateral part. These quantitative results support the current elastic fibre distribution scheme, and confirm the necessity of studying their orientation, taking into account age and temporomandibular joint health parameters.

Regional variations in the density and arrangement of elastic fibres in the anulus fibrosus of the human lumbar disc

Elastic fibres are critical components of the extracellular matrix in dynamic biological structures that undergo extension and recoil. Their presence has been demonstrated in the anulus fibrosus of the human lumbar intervertebral disc; however, a detailed regional analysis of their density and arrangement has not been undertaken, limiting our understanding of their structural and functional roles. In this investigation we have quantitatively described regional variations in elastic fibre density in the anulus fibrosus of the human L3-L4 intervertebral disc using histochemistry and light microscopy. Additionally, a multiplanar comparison of patterns of elastic fibre distribution in the intralamellar and interlamellar zones was undertaken. Novel imaging techniques were developed to facilitate the visualization of elastic fibres otherwise masked by dense surrounding matrix. Elastic fibre density was found to be significantly higher in the lamellae of the posterolateral region of the anulus than the anterolateral, and significantly higher in the outer regions than the inner, suggesting that elastic fibre density in each region of the anulus is commensurate with the magnitude of the tensile deformations experienced in bending and torsion. Elastic fibre arrangments in intralamellar and interlamellar zones were shown to be architecturally distinct, suggesting that they perform multiple functional roles within the anulus matrix structural hierarchy.

Dysregulated Apoptosis in Primary Varicose Veins

Objective Programmed cell death plays a critical role in various physiological processes. To investigate its possible pathogenic role in primary varicose veins we studied histological changes in surgical specimens from human varicose veins. In varicose and healthy veins, we also determined the number of cells in apoptosis, and investigated mediators regulating the intrinsic apoptotic mitochondrial pathway (Bax and caspase 9). Methods A total 23 varicose veins were obtained from 18 patients undergoing lower-extremity varicose vein surgery for primary varicose disorders. We used nine healthy veins obtained from nine patients undergoing distal arterial bypass grafting surgery as controls. The venous segment analysed was the distal part of the greater saphenous vein. Specimens for histological examination were stained with hematoxylin and eosin, trichromic and Victoria blue. Cell apoptoses and mediators of the mitochondrial pathway were detected in the media by immunohistochemistry using antibodies to peroxidase in situ apoptosis, Bax and caspase 9. Results were expressed as indexes for the three antibodies tested. The Mann–Whitney test was used to compare the results obtained in the two groups. Results Varicose vein specimens exhibited a more disorganised architecture than healthy veins and showed an increased number of collagen fibres and a decrease in the density and size of elastic fibres. All anti-apoptotic antibodies tested detected significantly fewer immunoreactive cells in tissue sections from the media of varicose veins than of healthy veins (peroxidase in situ, varicose veins (VV) median 2.4% (inter-quartile range 1.6–3.9) versus control (C) 14% (IQR 8.8–19); Bax, VV 1.4% (IQR 0.36–2.4) versus C 11% (IQR 7.6–15); and caspase 9, VV 1.7% (IQR 0.06–3.4) versus C 10% (IQR 9.1–12), P=0.0001 (Mann–Whitney test). Conclusion Apoptosis is down regulated in the medial layer of varicose veins. This dysregulation of the cellular mechanism that maintains normal tissue integrity is mediated through the intrinsic apoptotic pathway and may be among the causes of primary varicose veins.

Transplantation of reconstructed human skin on nude mice: a model system to study expression of human tenascin-X and elastic fiber components

Tenascin-X is a large extracellular matrix protein that is widely expressed in connective tissues during development and in the adult. Genetically determined deficiency of tenascin-X causes the connective tissue disease Ehlers-Danlos syndrome. These patients show reduced collagen density and fragmentation of elastic fibers in their skin. In vitro studies on the role of tenascin-X in elastic fiber biology are hampered because monolayers of fibroblasts do not deposit tenascin-X and elastic fibers into the extracellular matrix. Here, we applied an organotypic culture model of fibroblasts and keratinocytes to address this issue. We investigated the deposition of tenascin-X and elastin into skin-equivalent in vitro and also in vivo after transplantation onto immunodeficient mice. Whereas tenascin-C and fibrillin-1 were readily expressed in the skin-equivalents before transplantation, tenascin-X and elastin were not present. Three weeks post-grafting, a network of elastin was observed that coincided with the appearance of tenascin-X. At the ultrastructural level, microfibrils were observed, some of which were associated with elastin. Transplanted skin-equivalents containing tenascin-X-deficient fibroblasts showed deposition of immunoreactive elastin in similar quantities and distribution as those containing control fibroblasts. This suggests that tenascin-X is important for the stability and maintenance of established elastin fibers, rather than for the initial phase of elastogenesis. Thus, the transplantation of reconstructed skin on nude mice allows the study of tenascin-X and elastin expression and could be used as a model system to study the potential role of tenascin-X in matrix assembly and stability.

Volumetric density of elastic and reticular fibers in transition zone of controls and patients with benign prostatic hyperplasia

Objectives To analyze the volumetric density (Vv) of elastic and reticular fibers in the transition zone of controls and patients with benign prostatic hyperplasia (BPH). Methods Prostatic tissue samples were obtained from 25 patients (age range 63 to 79 years, mean 68) with clinical symptoms of bladder outlet obstruction who had undergone open prostatectomy. The control group was composed of 25 transition zone samples from prostates obtained during autopsy of adults aged younger than 30 years (killed in accidents). The Vv of the elastic and reticular fibers was determined in 25 random fields per prostate, using the point-count method with an M-42 grid test system. The data were analyzed using the Kolmogorov-Smirnov, Shapiro-Wilk W, and Mann-Whitney U tests. The Vv of these components was determined by stereologic methods. Results The Vv in the control and BPH groups was 12.47% ± 3.6% and 16.55% ± 9.11% in the elastic system fibers (difference not statistically significant) and 22.75% ± 1.66% and 29.33% ± 1.08% in the reticular fibers (difference statistically significant), respectively. Conclusions Reticular fibers showed a statistically significant increase in the BPH samples, and this finding suggests that stromal network fibers play a significant role in BPH.

The concentration of elastic fibres in the male urethra during human fetal development.

To describe the distribution of elastic fibres in the developing male urethra and to provide stereological data of the concentration of elastic fibres in the human urethra. Urethras were obtained from 10 fresh normal human fetuses at 15-36 weeks of gestation. A place-matched spongy urethra of a 27-year-old normal adult man was also analysed. Samples were fixed in Bouin's solution, embedded in paraffin and histologically processed. The elastic system fibres were evaluated by light microscopy using Weigert's resorcin-fuchsin technique after oxidation. Morphometric values were assessed by the point-counting method. The volumetric density (Vv) of elastic fibres was correlated with fetal age. At 15 weeks the elastic fibres were sparse and homogeneously distributed. The size and thickness of elastic fibres increased with age, mainly in the third trimester of gestation. Elastic fibres formed a randomly orientated network in the trabeculae of the corpus spongiosum. The mean (sem) Vv of elastic fibres in the spongy urethra was 5.2 (0.4)% in the fetus at 15 weeks and 14.8 (1.0)% at 36 weeks. In the urethra of the place- matched young man the Vv was 19.0 (1.3)%. The concentration of elastic fibres in the spongy urethra increased significantly with age. The high concentration of elastic fibres in the spongy urethra may partly explain its high extensibility. The progressive increase in elastic fibres during development implies functional adaptation of the fetal male urethra.

Effect of L-fucose and fucose-rich polysaccharides on elastin biosynthesis, in vivo and in vitro

With increasing age elastic fibres in human skin are progressively lysed and skin elasticity is also decreasing. Still there is an age-dependent increase of elastic fibre surface density, mostly due to an alteration of the fibres. The present experiments were undertaken to explore if L-fucose and fucose-rich polysaccharides (FROP-s) could influence elastin biosynthesis. We show here, that topical application of a fucose-containing preparation to the skin of hairless rats increased after 4 weeks the elastic fibre surface density by about 40%, shown by quantitative morphology. Using human skin fibroblasts in explant cultures, the addition of L-fucose or of FROP-3 increased the biosynthesis of immunoprecipitable tropoelastin by about 40%. No increase was found however of desmosine–isodesmosine in skin explant cultures after 72 h of incubation. The effect of L-fucose and FROP-3 on the biosynthesis of collagen and non-collagen proteins excreted by the skin explant cultures was also investigated. L-Fucose, but not FROP-3, decreased collagen biosynthesis but both increased non-collagen protein biosynthesis. These results show that L-fucose and FROP-3 stimulate tropoelastin biosynthesis in vitro, and elastic fibre formation in vivo. This stimulation concerns also several non-collagen proteins secreted by skin explant cultures. Elastic fibre formation necessitates the simultaneous synthesis of several microfibrillar glycoproteins as well as of tropoelastin. The increased elastic fibre density in the in vivo experiments suggests that this is indeed achieved by L-fucose and FROP-3, further demonstrating their efficiency in the control of age-dependent modifications of connective tissues in general and of skin in particular.

Estudio de fibras colágenas y elásticas del tejido conjuntivo de pacientes con y sin hernia inguinal primaria

There are few studies looking for collagen matrix defects in patients with inguinal bernia. To study the skin connective tissue in patients with and without inguinal bernia. Skin from the surgical wound was obtained from 23 patients with and 23 patients without inguinal bernia. The samples were processed for conventional light microscopy. Collagen fibers were stained with Van Giesson and elastic fibers with Weigert stain. Patients without hernia had compact collagen tracts homogeneously distributed towards the deep dermis. In contrast, patients with hernia had zones in the dermis with thinner and disaggregated collagen tracts. Connective tissue had a lax aspect in these patients. Collagen fiber density was 52% lower in patients with hernia, compared to subjects without hernia. No differences in elastic fiber density or distribution was observed between groups. Patients with inguinal bernia have alterations in skin collagen fiber quality and density.

Histomorphometric, biochemical and ultrastructural changes in the aorta of salt-loaded stroke-prone spontaneously hypertensive rats fed a Japanese-style diet

Background and Aim: It is demonstrated that dietary habits play a role in cardiovascular diseases. In stroke-prone spontaneously hypertensive rats (SHRsp), concomitant salt loading and a Japanese-style diet greatly accelerate hypertension and the appearance of cerebrovascular lesions by directly damaging arterial vessels. A number of studies have characterised medium and small vessel lesions in SHRsp, but little attention has been paid to the changes in the wall structure of large arteries induced by exposure to a salt-enriched diet. The aim of this study was to investigate the effects of a Japanese-style diet and salt loading on the thoracic aorta. Methods and Results: Two-month-old SHRsp were kept on a Japanese-style diet with 1% sodium chloride solution replacing tap water. Two months later, they were sacrificed and compared with age-matched or two-month-old control SHRsp kept on a standard diet and tap water in terms of the histomorphometry, ultrastructure and biochemical composition of the thoracic aorta. The vessel was consistently thicker in the four-month-old SHRsp (+20%, p<0.05 vs two-month-old rats) regardless of diet. The salt-loaded SHRsp showed a significant reduction in elastic fibre density (−20%, p<0.05 vs two-month-old rats) and an increase in the other matrix components (+50%), whereas the four-month-old controls showed preserved elastic fibres and a significant increase in the other matrix components (+65%, p<0.05 vs two-month-old rats). There was a considerable increase in the amounts of 4-OH-proline (+147%), 5-OH-lysine (+174%) and desmosines (+360%) in the four-month-old controls vs their two-month-old counterparts ( p<0.01), but not in the salt-loaded animals. Ultrastructural analysis revealed clear damage and accelerated aging in the thoracic aorta of the salt-loaded SHRsp. Conclusions: Salt loading and a Japanese-style diet destabilise thoracic aorta architecture in SHRsp after two months of treatment.

Maturation-dependent acquired coronary structural alterations and atherogenesis in the Dahl sodium-sensitive hypertensive rat.

The Dahl sodium-sensitive hypertensive rat exhibits atherogenic lesions after the initiation of a high-sodium/high-fat diet. This study was designed to gauge the effect of a preadolescent high-fat diet on the postadolescent rate of atherogenesis after supplementation of the diet with sodium. Fifty-three Dahl S male rats were assigned to 2 dietary groups for the postweaning to early adolescence period (3 to 12 weeks): 29 to a standard diet (low-fat/low-sodium) and 24 to a high-fat/low-sodium diet. At age 9 weeks (just after puberty), animals from the high-fat group exhibited a relatively diminished density of coronary elastic fibers. There was no evidence of either lipid or monocytic infiltration of the subendothelial space. At age 12 weeks, most or all of the remaining animals in both groups were switched to a high-sodium/high-fat diet and were sampled through the following 8 weeks for the appearance of arterial lipid. After the switch, the high-fat-conditioned animals developed more extensive atherosclerotic pathological lesions more rapidly than their prepubertal standard-diet counterparts. The importance of the animal's stage of maturation in this effect was underscored by the observation that delaying onset of the high-fat diet to early adolescence resulted in no ultimate difference from the pubertal controls in elastic fiber density. The maturation-dependent high-fat conditioning of these postweanling rats correlated with an accelerated rate of atherogenesis on the initiation of the high-sodium/high-fat diet, possibly as a direct result of an alteration in arterial elasticity.