Normal microscopic anatomy of equine body and limb skin: A morphological and immunohistochemical study

Jørgensen, Elin; Lazzarini, Giulia; Pirone, Andrea; Jacobsen, Stine; Miragliotta, Vincenzo

doi:10.1016/j.aanat.2018.03.010

Cited by 17 publications

(17 citation statements)

References 42 publications

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“…The location of mast cells in the subepidermal and deep dermal layers agrees with other studies conducted on equine skin (Fig. 3A-b) [17,18]. However, the scattered distribution within the granulation tissue of equine wounds has, to our knowledge, never been reported (Fig.…”

Section: Discussionsupporting

confidence: 91%

“…Mast cell characterization and quantification was performed by 2 blinded observers in the wound samples harvested from a former study [21]. In the intact skin borders of the wound, mast cell quantification was performed in the subepidermal layer (0-296 μm under basal membrane) and in the deep dermal layer (296-1184 μm) [18]. In the wound, mast cell quantification was performed in the granulation tissue.…”

Section: Image Analysis and Quantificationmentioning

confidence: 99%

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Development of a computer-based quantification method for immunohistochemically-stained tissues and its application to study mast cells in equine wound healing (proof of concept)

et al. 2020

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Background: There is a growing interest in the scientific community to use computer-based software programs for the quantification of cells during physiological and pathophysiological processes. Drawbacks of computer-based methods currently used to quantify immunohistochemical staining are the complexity of use, expense of software and overly-simplified descriptions of protocol thereby limiting reproducibility. The precise role of mast cells in equine cutaneous wound healing is unknown. Given the contribution of mast cells to the chronic inflammation observed in human keloid, a pathology similar to exuberant granulation tissue (EGT) in horses, mast cells might be present in high numbers in equine limb wounds predisposed to EGT. The main goal of this study was to develop a reliable and reproducible quantification method for immunostained tissues using a computer software that is widely available, at no cost, to the scientific community. A secondary goal was to conduct a proof of concept using the newly-established method to quantify mast cells during wound healing at different anatomical sites (body and limb) in horses to see if a different pattern is observed in limb wounds, which are predisposed to EGT. Results: A good intraclass correlation coefficient (ICC, 0.67 p < 0.05) was found between the computer-based ImageJ method and manual counting. An excellent intra-operator ICC of 0.90 (p < 0.01) was found for the ImageJ quantification method while a good interoperator ICC of 0.69 (p < 0.01) was measured. No significant difference was observed between the variation of the ImageJ and that of the manual counting method. Mast cells were localized below the epidermis, around cutaneous appendages and blood vessels. Mast cell numbers did not differ significantly in relation to anatomical location or time of healing. Conclusions: The computer-based quantification method developed is reliable, reproducible, available, cost-free and could be used to study different physiological and pathological processes using immunohistochemistry.

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Section: Discussionsupporting

confidence: 91%

Section: Image Analysis and Quantificationmentioning

confidence: 99%

Development of a computer-based quantification method for immunohistochemically-stained tissues and its application to study mast cells in equine wound healing (proof of concept)

et al. 2020

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“…Mast cell associations are highly variable as described above (Crane et al ; Sweeney et al ; Williamson et al ; Collobert‐Laugier et al ; Pittaway et al ; Hostetter et al ). Investigation of this other rare cell (Mair et al ; Jorgensen et al ) would be beneficial. Platelets are coming under increased scrutiny in human subjects and laboratory models for a broader role in immunomodulation of allergic reactions, particularly for their effect on eosinophil recruitment and function (Shah et al ).…”

Section: Discussionmentioning

confidence: 99%

Eosinophils of the horse: Part II: Eosinophils in clinical diseases

Brosnahan

2020

Equine Veterinary Education

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Interpretation of eosinophilia in body fluids or tissues is often not straightforward. Eosinophil counts vary among clinically healthy individuals, and considerable overlap can occur between normal and affected animals in conditions such as allergic airway disease. Parasite exposure is a confounding factor when counts are increased, and in cases where very high counts and dramatic clinical signs make another disease process obvious, the underlying pathology may be uncertain and treatment difficult. Eosinophils are a component of the immune response in many diseases of the horse, but their specific role is often unknown and likely multifactorial. In helminth infections, eosinophils are assumed to be part of the normal host response to a pathogen, whereas in multisystemic eosinophilic epitheliotropic disease (MEED), the predominance of eosinophils likely represents a wildly dysregulated response, or an abnormal response altogether. This distinction is still not clear for other diseases. Understanding the pathways involved in recruitment, activation or suppression of eosinophils is required for more accurate diagnostics, effective therapeutics, and successful strategies for prevention of eosinophil associated diseases. Eosinophils of the horse: Part II reviews published observations on the eosinophil in clinical diseases of the horse. The behaviour of eosinophils in three common and relatively wellstudied conditions is presented first, including gastrointestinal helminth infections, non-infectious respiratory disease, and insect bite hypersensitivity. The less common eosinophilassociated diseases such as eosinophilic disease confined to the intestine (EDCI) and MEED are considered, followed by a brief summary of the eosinophil in phycomycosis and neoplasia. In conclusion, a panoramic view of the equine eosinophil as presented in Parts I and II is placed in the larger context of current eosinophil research, and areas of study are identified that may improve our understanding of eosinophil biology in equine health and clinical disease. Gastrointestinal parasites of the horse Gastrointestinal helminths are a widespread and well-studied problem in the horse. Clinical disease is associated historically with migratory large strongyles (Strongylus vulgaris, S. edentatus, S. equinus), small strongyles (cyathostomes), tapeworms (Anoplocephala perfoliata) and, in foals, ascarids (Parascarus equorum). Early studies sought to define parasite life cycles, but post-infection necropsies also described the eosinophilic response (Duncan and Pirie 1975). Host immunity and anthelmintics were the focus of later works (Bailey et al. 1989; Steuer et al. 2018). Variables exist across studies, including dosing variations, natural versus experimental

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“…Sections (4 μm) were cut and deparaffinized using a standardized method by immersing the glass slides in xylene (3 × 5 min), 99% ethanol (2 × 5 min), 95% ethanol (2 × 5 min), 70% ethanol (1 × 5 min) and finally in distilled water (2 × 5 min). The spatial distribution of CK10, CK14, loricrin, PPAR‐α, E‐cadherin and N‐cadherin was investigated by indirect immunofluorescence.…”

Section: Methodsmentioning

confidence: 99%

“…Cytokeratin (CK)10, CK14 and loricrin are cytoskeletal proteins related to the differentiative state of keratinocytes and identification of these has been used to dissect the re‐epithelialization process in humans . In normal skin, CK10 and CK14 have been found in the suprabasal layers and basal layers of the epidermis, respectively . Loricrin is found in the granulosum layer of the epidermis and is the major structural protein in the cornified cell envelope.…”

Section: Introductionmentioning

confidence: 99%

Epithelial‐to‐mesenchymal transition and keratinocyte differentiation in equine experimental body and limb wounds healing by second intention

Jørgensen

Pirone

Jacobsen

et al. 2019

Veterinary Dermatology

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Background -The re-epithelialization process in equine wound healing is incompletely described. For epithelial cells to migrate during embryogenesis they undergo epithelial-to-mesenchymal transition (EMT); this phenotypic transition occurs during wound healing in humans and rodents, but it has not been investigated in horses.Hypothesis/objectives -To investigate keratinocyte differentiation and EMT in equine experimental excisional limb and body wounds healing by second intention. Animals -Six adult research horses.Methods and materials -Immunohistochemical analysis was used to detect expression of the differentiation markers cytokeratin (CK)10, CK14, loricrin and peroxisome proliferator-activated receptor alpha (PPAR-a), and of the EMT markers E-cadherin and N-cadherin in normal limb and body skin, and biopsies from limb and body wounds.Results -Loricrin and CK10 were expressed in normal skin and periwound skin but not in migrating epithelium of body and limb wounds. However, they reappeared at the migrating epithelial tip of body wounds only. CK14 and PPAR-a had uniform distribution throughout the migrating epithelium. N-cadherin was not expressed in normal unwounded skin but was detected in periwound skin adjacent to the wound margin. E-cadherin expression decreased at the wound margin.Conclusions and clinical importance -Presence of N-cadherin suggests that cadherin switching occurred during wound healing, this may be an indication that EMT occurs in horses. To the best of the authors' knowledge, this has never been described in horses before and warrants further investigation to assess the clinical implications. The tip of the migrating epithelium in body wounds appeared more differentiated than limb wounds, which could be part of the explanation for the superior healing of body wounds.

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Normal microscopic anatomy of equine body and limb skin: A morphological and immunohistochemical study

Cited by 17 publications

References 42 publications

Development of a computer-based quantification method for immunohistochemically-stained tissues and its application to study mast cells in equine wound healing (proof of concept)

Development of a computer-based quantification method for immunohistochemically-stained tissues and its application to study mast cells in equine wound healing (proof of concept)

Eosinophils of the horse: Part II: Eosinophils in clinical diseases

Epithelial‐to‐mesenchymal transition and keratinocyte differentiation in equine experimental body and limb wounds healing by second intention

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