Thick Elastic Fibers Research Articles

The characteristics of Laennec's capsule around the hepatic veins: A histological study based on 71 liver surgical specimens.

Laennec's capsule is a fibrous membrane attached to the surface of the liver, which is independent of the hepatic veins. However, the presence of Laennec's capsule surrounding the peripheral hepatic veins is controversial. This study aims to describe the characteristic of Laennec's capsule around the hepatic veins at all levels. Seventy-one hepatic surgical specimens were collected along the cross and longitudinal sections of the hepatic vein. Tissue sections of 3-4 mm were cut and stained with hematoxylin and eosin (H&E), resorcinol-fuchsin (R&F), and Victoria blue (V&B). Elastic fibers were observed around the hepatic veins. They were measured using K-Viewer software. Morphologically, we observed a thin, dense fibrous layer (so-called Laennec's capsule) around the hepatic veins at all levels, which was different from the thick elastic fibers of the hepatic vein wall. Therefore, there was a potential gap between Laennec's capsule and the hepatic veins. Laennec's capsule was visualized significantly better with R&F and V&B staining compared to H&E staining. The thickness of Laennec's capsule around the main, first, and secondary branches of the hepatic vein were 79.86 ± 24.20 μm, 48.41 ± 18.25 μm, and 23.56 ± 10.03 μm in the R&F staining, and 80.15 ± 21.85 μm, 49.46 ± 17.52 μm, and 25.05 ± 11.03 μm in the V&B staining, respectively. They were significantly different from each other (P < .001). The hepatic veins were surrounded by Laennec's capsule at all levels, including the peripheral hepatic veins. However, it is thinner along the vein branches. The gap between the Laennec's capsule and hepatic veins shows potential supplemental value for liver surgery.

The Potential Use of Elastic Tissue Autofluorescence in Formalin-fixed Paraffin-embedded Skin Biopsies

Autofluorescence (AF) or naïve-florescence is the natural emission of light by biomolecules. During florescence microscope examination, we realized that elastic tissue is brighter or more autoflourescent than collagen and other biomolecules/cells in the skin. Consequently, we decided to review elastic tissue-related pathologies under a florescence microscope and to report the possible benefits of this technique from selected cases from the paraffin-block archive, by using the protease digestion immunofluorescence method. Selected and clinic-pathologically confirmed 3 elastofibroma dorsi, 3 pseudoxanthoma elasticum, 3 anetoderma, 3 arteriovenous malformations, 3 temporal arteritis, 3 scar tissue and 3 highly solar-damaged samples of skin from 2014-2019 were retrieved. Under the fluorescent microscope, coarse, thick and globularly-fragmented elastic fibers of elastofibroma dorsi, shortened, irregular and convoluted elastic fibers of pseudoxanthoma elasticum, internal elastic membranes of arteries and their integrity was visualized. None of the anetoderma cases had any signal representing elastic tissue. It was shown that elastic tissue can be observed easily under fluorescence microscope in the case of FFPE tissues. The resulting autofluorescence can be useful in recognizing elastic tissue-related pathologies, and it may be used as an ancillary or an alternative method to routine histochemical techniques.

Effects of different intensity exercise on morphology and structure of atrioventricular node and protein expressions of VEGF, VIP and Syn in rats

Objective: To investigate the effects of different intensities of aerobic exercise on the morphology of the atrioventricular node and the expressions of vascular endothelial growth factor (VEGF), vasoactive intestinal peptide (VIP) and synaptophysin (Syn) in rats. Methods: Three-month-old SD rats were randomly divided into 3 groups: quiet control group (C), moderate-intensity aerobic exercise group (AE), and high-intensity aerobic exercise group (FE), each with 8 rats. The moderate-intensity aerobic exercise and high-intensity aerobic exercise rat models were rats established by using 8-week treadmill training ways. Results: Compared with the quiet control group, in the moderate-intensity aerobic exercise group, the atrioventricular nodule muscle fibers were evenly distributed, with smaller gaps, a sense of transparency, and collagen tightly wraps around the atrioventricular nodules. The intercalary disc had a clear structure and complete connection, and the capillary tube wall was relatively clear. Thick elastic fibers were more developed, the expression levels of VEGF and VIP in the atrioventricular node were increased significantly (P＜0.05), and there was no significant change in the expression of Syn. In the high-intensity aerobic exercise group, the atrioventricular nodule muscle fibers were unevenly distributed and arranged abnormally, connection between cells were disordered, and vacuolated. The intercalary disk was twisted, circling and overlapping, fuzzy, discontinuous, and extremely dilated. The wall was thick and the endothelial cells were arranged sparsely and disorderly. Moreover, high-intensity aerobic exercise significantly inhibited the expressions of VEGF, VIP, and Syn (P＜0.05). Conclusion: Aerobic exercise training of different intensities has obvious effects on the morphology and structure of the atrioventricular node and the expressions of VEGF, VIP and Syn. Moderate aerobic exercise can maintain and promote the normal morphology and structure of the atrioventricular node, while high-intensity aerobic exercise can damage the morphology and structure of the ventricular node, which is presumably closely related to the arrhythmia induced by high-intensity exercise.

The ultrastructural organization of elastic fibers at the interface of the nucleus and annulus of the intervertebral disk

There has been no study to describe the ultrastructural organization of elastic fibers at the interface of the nucleus pulposus and annulus fibrosus of the intervertebral disk (IVD), a region called the transition zone (TZ). A previously developed digestion technique was optimized to eliminate cells and non-elastin ECM components except for the elastic fibers from the anterolateral (AL) and posterolateral (PL) regions of the TZ in ovine IVDs. Not previously reported, the current study identified a complex elastic fiber network across the TZ for both AL and PL regions. In the AL region, this network consisted of major thick elastic fibers (≈ 1 µm) that were interconnected with delicate (< 200 nm) elastic fibers. While the same ultrastructural organization was observed in the PL region, interestingly the size of the elastic fibers was smaller (< 100 nm) compared to those that were located in the AL region. Quantitative analysis of the elastic fibers revealed significant differences in the size (p < 0.001) and the orientation of elastic fibers (p = 0.001) between the AL and PL regions, with a higher orientation and larger size of elastic fibers observed in the AL region. The gradual elimination of cells and non-elastin extracellular matrix components identified that elastic fibers in the TZ region in combination with the extracellular matrix created a honeycomb structure that was more compact at the AF interface compared to that located close to the NP. Three different symmetrically organized angles of rotation (0⁰ and ±90⁰) were detected for the honeycomb structure at both interfaces, and the structure was significantly orientated at the TZ-AF compared to the TZ-NP interface (p = 0.003).

Elastic fibers: The missing key to improve engineering concepts for reconstruction of the Nucleus Pulposus in the intervertebral disc

The increasing prevalence of low back pain has imposed a heavy economic burden on global healthcare systems. Intense research activities have been performed for the regeneration of the Nucleus Pulposus (NP) of the IVD; however, tissue-engineered scaffolds have failed to capture the multi-scale structural hierarchy of the native tissue. The current study revealed for the first time, that elastic fibers form a network across the NP consisting of straight and thick parallel fibers that were interconnected by wavy fine fibers and strands. Both straight fibers and twisted strands were regularly merged or branched to form a fine elastic network across the NP. As a key structural feature, ultrathin (53 ± 7 nm), thin (215 ± 20 nm), and thick (890 ± 12 nm) elastic fibers were observed in the NP. While our quantitative analysis for measurement of the thickness of elastic fibers revealed no significant differences (p < 0.633), the preferential orientation of fibers was found to be significantly different (p < 0.001) across the NP. The distribution of orientation for the elastic fibers in the NP represented one major organized angle of orientation except for the central NP. We found that the distribution of elastic fibers in the central NP was different from those located in the peripheral regions representing two symmetrically organized major peaks (±45⁰). No significant differences in the maximum fiber count at the major angles of orientation (±45⁰) were observed for both peripheral (p = 0.427) and central NP (p = 0.788). Based on these new findings a structural model for the elastic fibers in the NP was proposed. The geometrical presentation, along with the distribution of elastic fibers orientation, resulting from the present study identifies the ultrastructural organization of elastic fibers in the NP important towards understanding their mechanical role which is still under investigation. Given the results of this new geometrical analysis, more-accurate multiscale finite element models can now be developed, which will provide new insights into the mechanobiology of the IVD. In addition, the results of this study can potentially be used for the fabrication of bio-inspired tissue-engineered scaffolds and IVD models to truly capture the multi-scale structural hierarchy of IVDs. Statement of SignificanceVisualization of elastic fibers in the nucleus of the intervertebral disk under high magnification was not reported before. The present research utilized extracellular matrix partial digestion to address significant gaps in understanding of nucleus microstructure that can potentially be used for the fabrication of bio-inspired tissue-engineered scaffolds and disk models to truly capture the multi-scale structural hierarchy of discs.

Three-dimensional analysis of alveolar wall destruction in the early stage of pulmonary emphysema.

The morphological mechanism of alveolar wall destruction during pulmonary emphysema has not been clarified. The aim of this study was to elucidate this process three-dimensionally. Lung specimens from five patients with pulmonary emphysema were used, and five controls with normal alveolar structure were also examined. Sections 150 μm thick were stained with hematoxylin and eosin, elastica, and silver impregnation, and immunostained with selected antibodies. We examined these sections three-dimensionally using a laser confocal microscope and a light microscope. There were only a few Kohn's pores and no fenestrae in the normal alveoli from the controls. In the lungs of the emphysema patients a small rupture appeared in the extremely thin alveolar wall among the alveolar capillaries. This rupture enlarged to form a circle surrounded by the capillaries, which was called an alveolar fenestra. Two neighboring fenestrae fused by breakdown of the collapsed or cord-like capillary between them to form a large fenestra. The large fenestrae fused repeatedly to become larger, and these were bordered by thick elastic fibers constructing an alveolar framework. Alveolar wall destruction during emphysema could start from small ruptures of the alveolar wall that become fenestrae surrounded by capillaries, which fuse repeatedly to become larger fenestrae rimmed with elastic fibers. The alveolar capillary network could initially prevent enlargement of the fenestrae, and the thick elastic fibers constituting the alveolar framework could secondarily prevent destruction of the alveolar wall structure.

Elastic fiber pattern in regressing melanoma: a histochemical and immunohistochemical study

Although histopathologic identification of regression of melanoma is usually straightforward, sometimes it can be difficult to distinguish it from scarring fibrosis. Therefore, this study investigates the elastic fiber pattern in melanomas associated with either regression or scars. We compared 33 invasive melanomas with the fibrosing stage of regression to 10 cases of invasive melanomas with scarring fibrosis. None of the regression cases had a prior surgical procedure. Elastic fiber patterns were evaluated with Verhoeff's elastic van Gieson stain (EVG) and elastin immunostain. Elastin immunostain was superior to EVG in revealing the elastic fiber patterns. Both regression and scars had decreased to absent elastic fibers in the areas of fibrosis. However, areas of regression had a well-defined compressed layer of thin elastic fibers pushed down from the papillary dermis to the base of the fibrosis. In contrast, the base of scars lacked this compressed elastic layer and had instead an abrupt transition to the thick elastic fibers of the spared reticular dermis. We have identified distinct changes of the elastic tissue network, which more accurately define the presence of regression in melanoma and distinguish it from scarring fibrosis.

Choroid plexus papilloma with stromal deposition of amyloid and elastic material

Congophilic birefringent amyloid deposits, with immunostaining for transthyretin (TTR) and amyloid P, associated with numerous coarse, enlarged and thick elastic fibres, are reported in the stroma of two choroid plexus papillomas, a finding not previously described in choroid plexus tumours. TTR was expressed as aggregates of ‘doughnut-shaped’ bodies, in which the TTR-positive peripheral area encircled the elastic fibre (TTR-negative core). Ultrastructurally, the amyloid microfibrils surrounded the elastic fibres and appeared to continue into the microfibrillar mantle of the latter. The stromal TTR-amyloid deposits associated with abundant elastic fibres in tumours that occur in the choroid plexus may be related to the alteration (production/accumulation, insufficient breakdown and/or extracellular matrix modifications) of some of the choroid plexus functions (removal, target and source of polypeptides, including TTR synthesis) and may be of interest for future studies on choroid plexus polypeptide activity and on protein development into elastomeric and amyloidogenic microfibrils.

Histological investigation of rabbit ligamentum flavum with special reference to differences in spinal levels.

The structure of the ligamentum flavum has yet to be fully elucidated and no studies have investigated fine structural differences at different spinal levels in any animals. The aim of the present study was to clarify structural differences in the ligamentum flavum at different spinal levels (cervical: C3/4 and C5/6; upper thoracic: T2/3; lower thoracic: T9/10; lumbar: L3/4) using light and electron microscopy of rabbit specimens. Light microscopy using resorcin-fuchsin staining revealed that the distribution of elastic fibers was diffuse in the cervical and upper thoracic regions, but was generally dense in the lower thoracic and lumbar regions. Electron microscopy demonstrated that the cervical and upper thoracic regions were rich in collagen fibers. Conversely, the lower thoracic and lumbar regions were rich in elastic fibers. Quantitative image analyses displayed thick elastic fibers in the lower thoracic and lumbar regions, with high area ratios. Radiographic examinations revealed that ranges of motion were large at the cervical region, but small at the lower thoracic and lumbar regions. These findings suggest that structure of the ligamentum flavum varies at different spinal levels with respect to differences in motion.

Innervation and functional characteristics of connective tissues, especially elastic fibers, in human fetal thoracic intervertebral articular capsule and its surroundings.

The articular capsules between the thoracic vertebrae, which have physiologically different functions from those of other levels of the vertebrae, have yet to be subjected to neuro-anatomical and fine structural analysis. In the present study, we analyzed serial frozen sections of decalcified thoracic vertebrae in human fetuses, and identified the articular capsule tissue with its unique distribution of elastic fibers. The fine structure of the elastic fibers was studied by transmission electron microscopy. In the early-stage fetus, the fibrous membrane forming the lateral intervertebral articular capsule contained abundant thin elastic fibers consisting of microfibrils. In the late-stage fetus, the lateral capsule of fibrous membrane was occupied by thick elastic fibers. A medial articular capsule, namely the ligamenta flava, contained numerous thick elastic fibers in both early and late-stage fetuses. The distributional differences in nerve fibers between early and late-stage fetuses were determined by immunostaining, using antibodies raised against protein gene product 9.5 (PGP 9.5; ubiquitin carboxyl-terminal hydrolase). Innervation by PGP 9.5 immunoreactive fibers was limited to the areas of the articular capsules near the blood vessels, which may indicate their functional relation with blood flow. No PGP 9.5 immunoreactive fibers were found in the ligamenta flava of the late-stage fetus. Innervation might be directly involved in the development of the intervertebral articular capsules in normal human fetuses.

Round window membrane in young and aged C57BL/6 mice

Although there have been many studies on the round window membrane (RWM), little information has been reported about changes in the membrane associated with aging. We have undertaken morphological studies of RWMs using young (7–8 weeks old) and aged (27–29 months old) C57BL/6 mice. The RWM was thinner in mice from the aged group compared with that of the young group. The cell density in the epithelial and inner layers was also reduced in the aged group. In the middle layer of the RWM in aged mice, transmission electron microscopy revealed many degenerated short and thick elastic fibers. Confocal laser microscopy using fluorescein isothiocyanate(FITC)–wheat germ agglutinin (WGA) staining was used to identify WGA-positive fibers in the middle layer of the RWM, which changed in a similar manner to the fibers in aging skin.

Three-dimensional analysis of alveolar structure in usual interstitial pneumonia

The etiology of usual interstitial pneumonia (UIP), a progressive lung disease, remains unclear. We examined alveolar structure in UIP three-dimensionally. Lung biopsy specimens from five patients with idiopathic pulmonary fibrosis were used. Sections 150-microm thick were stained with elastica solution for elastic fibers, with alpha-smooth muscle actin antibody for myofibroblasts, with anti-Thomsen-Friedenreich antibody for type-II pneumocytes and with anti-CD34 antibody for blood vessels. We examined them three-dimensionally using a laser confocal microscope or light microscope. In the fibrotic lesions, the thick elastic fibers forming the alveolar framework were not particularly dense considering the reduction in alveolar volume. Near the fibrotic lesions, some of the thin elastic fibers in the alveolar wall were slightly sinuous and ended with rounded tips. Type-II pneumocytes had proliferated and were distributed uniformly over the alveolar surface. Smooth muscle actin filaments were detected only around the alveolar orifice. These findings show that in UIP destruction of the elastic fiber framework of the alveoli may lead to irreversible focal alveolar collapse after damage to the alveolar epithelial cells, and proliferation of type-II pneumocytes may be involved with this elastolysis.

Variation of heads of lateral pterygoid muscle and morphology of articular disc of human temporomandibular joint--anatomical and histological analysis.

To grasp the documented evidences available regarding the cause of the anteriorly displaced human temporomandibular joint (TMJ) disc. With 20 specimens of human TMJ from cadavers, at first the number of heads of lateral pterygoid muscle (LPM) bundles were analysed with macroanatomical method. Secondary histological procedure with haematoxyline and eosine staining and orceine staining were carried out for the same specimens. Macroscopically, in 13 specimens an upper and a lower head of the LPM were present. In the other seven specimens, not only an upper and a lower head, but also a third inner head of the LPM covered with a separate fascia was observed. Histochemically, the specific localization of certain bundles of thick elastic fibres in the antero-medial parts of the anteriorly displaced deformed TMJ disc were observed.

Type II pneumocytes are preferentially located along thick elastic fibers forming the framework of human alveoli.

Type II pneumocytes were found to be preferentially located on thick elastic fibers which formed the main structural framework of the alveoli in humans. Eight lobes resected from eight patients with adenocarcinoma or atypical adenomatous hyperplasia and two lobes from two necropsies were examined. Four small specimens of normal lung tissue were obtained from each lobe fixed in a buffered formalin. Thick sections (200-300 microm) were stained with hematoxylin to contrast nuclei or with elastica staining to demonstrate elastic fibers and immunostained with an antibody against Thomsen-Friedenreich antigen after pretreatment with sialidase to visualize type II pneumocytes. Alveolar structure and the distribution of type II pneumocytes were examined in 3D reconstructions generated using a standard microscope, a personal computer and NIH-Image software. Nuclei including those of type I and type II pneumocytes and endothelial cells were distributed diffusely throughout the alveolar wall. Thick elastic fibers constructed the main structural framework of the alveoli and formed the sides of the polygonal alveoli. Type II pneumocytes were found to be linearly located along these thick elastic fibers. This previously undisclosed distribution of type II pneumocytes may be concerned with alveolar rapid movement.

Type II pneumocytes are preferentially located along thick elastic fibers forming the framework of human alveoli

Type II pneumocytes were found to be preferentially located on thick elastic fibers which formed the main structural framework of the alveoli in humans. Eight lobes resected from eight patients with adenocarcinoma or atypical adenomatous hyperplasia and two lobes from two necropsies were examined. Four small specimens of normal lung tissue were obtained from each lobe fixed in a buffered formalin. Thick sections (200–300 μm) were stained with hematoxylin to contrast nuclei or with elastica staining to demonstrate elastic fibers and immunostained with an antibody against Thomsen-Friedenreich antigen after pretreatment with sialidase to visualize type II pneumocytes. Alveolar structure and the distribution of type II pneumocytes were examined in 3D reconstructions generated using a standard microscope, a personal computer and NIH-Image software. Nuclei including those of type I and type II pneumocytes and endothelial cells were distributed diffusely throughout the alveolar wall. Thick elastic fibers constructed the main structural framework of the alveoli and formed the sides of the polygonal alveoli. Type II pneumocytes were found to be linearly located along these thick elastic fibers. This previously undisclosed distribution of type II pneumocytes may be concerned with alveolar rapid movement. Anat Rec 258:34–38, 2000. © 2000 Wiley-Liss, Inc.

Linear focal elastosis: Keloidal repair of striae distensae

Linear focal elastosis and striae distensae coexisted in a 32-year-old black man in their typical distribution on the lower back. Many striae appeared when he lost 50 lb in 1991. Only some of them became hypertrophic and others remained atrophic; striae distensae of the shoulders and buttocks remained atrophic, whereas most in the lumbosacral region became indurated and cordlike. Histologically, the former lacked elastic fibers and the latter showed a marked increase in thick elastic fibers. Linear focal elastosis in some patients may represent hyperplasia of elastic fibers in response to their damage. This “elastic fiber keloid” seems to be equivalent to collagen keloid, i.e., ordinary keloid, in the lumbosacral region. (J Am Acad Dermatol 1998;39:309-13.)

Microfibrillar elements of the dermal matrix.

Connective tissue microfibrils are key structural elements of the dermal matrix which play major roles in establishing and maintaining the structural and mechanical integrity of this complex tissue. Type VI collagen microfibrils form extensive microfibrillar networks which intercalate between the major collagen fibrils and are juxtaposed to cellular basement membranes, blood vessels and other interstitial structures. Fibrillin microfibrils define the continuous elastic network of skin, and are present in dermis as microfibril bundles devoid of measureable elastin extending from the dermal-epithelial junction and as components of the thick elastic fibres present in the deep reticular dermis. Electron microscopic analyses have revealed both classes of microfibrils to have complex ultrastructures. The ability to isolate intact native microfibrils from skin has enabled a combination of high resolution and biochemical techniques to be applied to elucidate their structure:function relationships. These approaches have generated new information about their molecular organisation and physiological interactions in health and disease.

Identification of elastic fibres in the peripheral nerve

Traditional histological staining techniques, as well as elastin-specific antibodies and electron microscopy, have been used to assess the distribution of elastin within the peripheral nerve. The location of the elastin identified by the VerHoeff VanGiesen or Weigert stains has been shown to coincide with the unambiguous identification of elastin by immunospecific stains and electron microscopy. Elastin is located in all three connective layers of the peripheral nerve. Thick elastic fibres, consisting of amorphous elastin protein and microfibrils, are located consistently in the perineurium and, to a lesser extent, in the epineurium. The endoneurium contains small collections of elastic fibres widely distributed between the axons. Compared with collagen, the overall content of elastin, however, is small, suggesting that the visco-elastic properties of peripheral nerve may be due primarily to collagen.

Histogenesis of abnormal elastic fibers in blebs and bullae of patients with spontaneous pneumothorax: ultrastructural and immunohistochemical studies.

Elastic fibers in 15 blebs and 17 bullae with spontaneous pneumothorax were studied by means of electron microscopy and light and electron microscopic immunohistochemistry for elastin and alpha 1-antitrypsin. Blebs were formed in association with focal organized alveoli, and bullae were formed in association with pulmonary emphysema. Both blebs and bullae had abnormal elastic fibers. Ultrastructurally, abnormal elastic fibers of blebs and bullae consisted of accumulated thick and fine fibers. Accumulated thick elastic fibers showed vacuolar changes and electron-dense granular deposits, and they were associated with spiraling collagen fibrils. These thick elastic fibers reacted evenly with antielastin antibody and also reacted with anti-alpha 1-antitrypsin antibody. They are thought to be degraded elastic fibers. Accumulated fine elastic fibers consisted of bundles of microfibrils and granular amorphous components, and they reacted with anti-elastin and anti-alpha 1-antitrypsin antibody. These fine elastic fibers are thought to be not only newly formed in the process of organization but also degraded. It is suggested that elastic fibers of blebs and bullae are degraded due to an imbalance between elastase and alpha 1-antitrypsin.

Changes in Extracellular Matrix Materials in the Uterine Myometrium of Rats during Pregnancy and Postparturition

Morphological changes in extracellular matrix materials in the uterine myometrium of rats during pregnancy and postparturition were studied by light and electron microscopy together with immunofluorescence microscopy for type III and IV collagens, fibronectin and laminin. The main components present in late pregnancy were 1) various-sized collagen fibrils, 2) thick elastic fibers adjacent to smooth muscle cells, and 3) continuous and thick basement membranes of hypertrophic smooth muscle cells. These findings are considered to indicate degradation of collagen fibrils and development of elastic fibers and basement membranes of smooth muscle cells. This change in extracellular matrix materials in the late stage of pregnancy may be important in the process of uterine enlargement associated with elasticity and preparation for labor. In the postpartum stage, myofibroblastic interstitial cells were seen to phagocytize collagen fibrils, and elastic fibers accumulated mainly around the bundles of smooth muscle cells. These changes in the postpartum stage are thought to be important for the process in which the uterus returns to the nonpregnant condition. It is suggested that smooth muscle cells participate in regulating the development of their basement membranes and elastic fibers, and that myofibroblastic interstitial cells function by clearing degraded collagen fibrils from the uterine myometrium.