Segmentation of elastic fibres in images of vessel wall sections stained with Weigert's resorcin–fuchsin

Hernndez-Morera, Pablo; Travieso-Gonzlez, Carlos; Castao-Gonzlez, Irene; Mompe-Corredera, Blanca; Ortega-Santana, Francisco

doi:10.1016/j.cmpb.2017.02.018

Cited by 8 publications

(4 citation statements)

References 20 publications

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“…In a similar manner to that observed in elastic fibre volume, a smaller area of elastic fibres (AFE) was also identified among animals in the GCT group compared to the other treatment groups. Based on the equivalence of the statistical analysis between AFE and VEF and the brevity of time (12 days) used to evaluate the elastic fibre formation, the same biological hypotheses discussed previously for VEF are likely to persist; however, the colour segmentation technique has a low applicability for measuring elastic fibres in different tissues [12]. To date, no studies have been reported that apply this technique to the analysis of dermal elastic fibres.…”

Section: Discussionmentioning

confidence: 99%

“…The scientific literature describes the colour segmentation technique used in the present study in detail, including in several contemporary studies that have assessed the applicability of K-means clustering in dermatological research [5,4,12]. This study opted for the target colour, as binarisation methods commonly employed in morphology can overestimate the values resulting from the absence of sample categorisation in different groups [18].…”

Section: Discussionmentioning

confidence: 99%

“…To date, no studies have been reported that apply this technique to the analysis of dermal elastic fibres. One recent study developed a specific algorithm for measuring elastic fibres in blood vessels [12], which indicates that this technique is still under development.…”

Section: Discussionmentioning

confidence: 99%

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Stereological and Morphometric Study of Type 3 Collagen Formation in the Cutaneous Wounds of Diabetic Mice Treated with Mesenchymal Stem Cells

Rodrigues

Neto

Silva

et al. 2018

Acta Scientiae. Vet.

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Introduction: Wound healing is a progressive, essential and complex physiological process that occurs as a restorative response after a tissue injury. It involves three phases: inflammation, proliferation and maturation. Exogenous, endogenous and pathological factors may interfere in the cicatricial process in humans and animals by altering the balance between the synthesis, degradation and remodelling of collagen and elastic fibres. Diabetes mellitus is a progressive metabolic disease that alters elastogenesis and collagenesis and induces delays in the healing process. Scientific evidence suggests that mesenchymal stem cells modulate the cicatricial response. Thus the objective of this work was to perform stereological and morphometric analysis to determine the formation of dermal fibres in cutaneous fragments of a murine model of diabetes mellitus.Materials, Methods & Results: Histological sections were obtained from the cutaneous wounds of diabetic mice. The cutaneous wounds were previously treated with autogenous mesenchymal stem cells, physiological solution or polyurethane membrane. The histological sections were subsequently processed and stained for type 1 and 3 collagen fibres and elastic fibres using Picrosirius Red and Weigert staining, respectively. Histological sections stained with Picrosirius Red presented three types of birefringence under polarised light microscopy that corresponded to red colours for type 1 collagen and green and yellow colours for type 3 collagen. Weigert staining presented three colours for histological structures under white light microscopy that corresponded to black colours for elastic fibres, variations in colour from pink to purple for other structures and dermal attachments. The elastic fibres, represented by a black colour, presented in a heterogeneous form and were either identified as thin, punctiform or rectangular fibres or as elastic agglomerates. A greater volume of elastic fibres was observed in the superficial dermis than in the deep dermis, arranged irregularly. These fibres were organised longitudinally to the dermo-epidermal junction and surrounding the blood vessels and hair follicles. The images obtained were evaluated using the Cavalieri principle of stereology to obtain quantitative data in three-dimensions (3D), represented by the volume of the dermal fibres, and by the colour segmentation method. The K-means clustering plug-in in Image J® was used to quantify the area of the dermal fibres in the cutaneous wounds after the proposed dermatological treatments. A total of 90 images were obtained and analysed. No statistically significant differences (P > 0.01) were observed in the volume or area of type 1 collagen fibres between the treatment groups. Significant differences (P < 0.01) were only identified for the volumes and areas of type 3 collagen, with treated animals also presenting lower mean values for the volume and area of elastic fibres compared to the control group.Discussion: The preponderance of type 3 immature collagen in the cutaneous wounds of animals treated with stem cells indicates active collagenase and greater fibroblastic activity, which is probably induced by stem cells. Diametrically, the identification of lower levels of elastic fibres in the cutaneous fragments treated with stem cells suggests that cell therapy does not contribute satisfactorily to elastogenesis. Previous reports suggested that mesenchymal stem cells may decrease elastin synthesis, and such a situation may have occurred in this study. The autologous mesenchymal stem cells increased the formation of collagen fibres in diabetic mice at the detriment of the formation of elastic fibres, thus suggesting active early collagen in the first 2 weeks of the cicatricial process.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

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Stereological and Morphometric Study of Type 3 Collagen Formation in the Cutaneous Wounds of Diabetic Mice Treated with Mesenchymal Stem Cells

Rodrigues

Neto

Silva

et al. 2018

Acta Scientiae. Vet.

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“…WRM can dye the elastic fibers dark blue, making the widely distributed elastic fibers in the walls of alveoli and blood vessels be visualized ( Proctor and Horobin, 1988 ). Both staining techniques are widely used in histological and pathological studies ( Hochstim et al, 2010 ; Hernandez-Morera et al, 2017 ). In this study, three and one samples per group were used for HE and WRM staining, respectively; the samples were fixed with 4% paraformaldehyde, then dehydrated in graded ethanol (75%→85%→95%→100%), rinsed in xylol, and embedded in paraffin.…”

Section: Methodsmentioning

confidence: 99%

Physiology and Transcriptomics Analysis Reveal the Contribution of Lungs on High-Altitude Hypoxia Adaptation in Tibetan Sheep

Zhao

et al. 2022

Front. Physiol.

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The Tibetan sheep is an indigenous species on the Tibetan plateau with excellent adaptability to high-altitude hypoxia and is distributed at altitudes of 2500–5000 m. The high-altitude hypoxia adaptation of Tibetan sheep requires adaptive reshaping of multiple tissues and organs, especially the lungs. To reveal the mechanisms of adaptation at the tissue and molecular levels in the lungs of Tibetan sheep under hypoxic conditions at different altitudes, we performed light and electron microscopic observations, transcriptomic sequencing, and enzyme-linked immunosorbent assay studies on the lungs of Tibetan sheep from three altitudes (2500, 3500, and 4500 m). The results showed that in addition to continuous increase in pulmonary artery volume, thickness, and elastic fiber content with altitude, Tibetan sheep increase the hemoglobin concentration at an altitude of 3500 m, while they decrease the Hb concentration and increase the surface area of gas exchange and capacity of the blood at an altitude of 4500 m. Other than that, some important differentially expressed genes related to angiogenesis (FNDC1, HPSE, and E2F8), vasomotion and fibrogenesis (GJA4, FAP, COL1A1, COL1A2, COL3A1, and COL14A1), and gas transport (HBB, HBA1, APOLD1, and CHL1) were also identified; these discoveries at the molecular level explain to some extent the physiological findings. In conclusion, the lungs of Tibetan sheep adopt different strategies when adapting to different altitudes, and these findings are valuable for understanding the basis of survival of indigenous species on the Tibetan plateau.

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The sphenoidal emissary foramen and the emissary vein: Anatomy and clinical relevance

et al. 2019

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Although the sphenoidal emissary foramen (SEF) and its content are anatomically and clinically relevant, accurate description of them in the modern literature is lacking. This study aimed to examine and describe the SEF and its content (the sphenoidal emissary vein [SEV]). We analyzed 1,000 computed tomography (CT) images, 170 dry skulls, 50 formalin‐fixed specimens, and three specimens (heads) following guidelines proposed by Dr. Albert L. Rhoton Jr. MD for latex injection. SEV morphology was determined by histological staining and electron microscopy. The SEF was observed in 46.8% of the CTs studied (25.4% bilateral and 21.4% unilateral), and 45.2% of the dry skulls (18.8% bilateral and 26.4% unilateral). In 9.5% of CTs and 21.1% of dry skulls there was a blind channel in the external surface of the cranial base; since there was no communication with the cranial cavity, it was not considered as the SEF. During the dissections, the SEF was found in seven individuals. In three of them, the SEV was an alternative route for venous drainage of the venous plexus of the foramen ovale. Its walls were composed of collagen fibers and its endothelium contained rhomboid cells resembling those commonly found in the superior sagittal sinus. The presence of the SEF and SEV can anatomically explain the spread of certain cranial base pathologies from or toward Meckel's cave or the cavernous sinus, and should be taken into account during procedures in the middle cranial fossa, percutaneous approaches, odontological procedures, and treatment of dural arteriovenous fistulas. Clin. Anat., 33:767–781, 2020. © 2019 Wiley Periodicals, Inc.

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Segmentation of elastic fibres in images of vessel wall sections stained with Weigert's resorcin–fuchsin

Cited by 8 publications

References 20 publications

Stereological and Morphometric Study of Type 3 Collagen Formation in the Cutaneous Wounds of Diabetic Mice Treated with Mesenchymal Stem Cells

Stereological and Morphometric Study of Type 3 Collagen Formation in the Cutaneous Wounds of Diabetic Mice Treated with Mesenchymal Stem Cells

Physiology and Transcriptomics Analysis Reveal the Contribution of Lungs on High-Altitude Hypoxia Adaptation in Tibetan Sheep

The sphenoidal emissary foramen and the emissary vein: Anatomy and clinical relevance

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