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Article: Myelin histology: a key tool in nervous system research.

García-García, Óscar Darío / Carriel, Víctor / Chato-Astrain, Jesús

2023 Volume 19, Issue 2, Page(s) 277–281

Abstract: The myelin sheath is a lipoprotein-rich, multilayered structure capable of increasing conduction velocity in central and peripheral myelinated nerve fibers. Due to the complex structure and composition of myelin, various histological techniques have been ...

Abstract	The myelin sheath is a lipoprotein-rich, multilayered structure capable of increasing conduction velocity in central and peripheral myelinated nerve fibers. Due to the complex structure and composition of myelin, various histological techniques have been developed over the centuries to evaluate myelin under normal, pathological or experimental conditions. Today, methods to assess myelin integrity or content are key tools in both clinical diagnosis and neuroscience research. In this review, we provide an updated summary of the composition and structure of the myelin sheath and discuss some histological procedures, from tissue fixation and processing techniques to the most used and practical myelin histological staining methods. Considering the lipoprotein nature of myelin, the main features and technical details of the different available methods that can be used to evaluate the lipid or protein components of myelin are described, as well as the precise ultrastructural techniques.
Language	English
Publishing date	2023-07-23
Publishing country	India
Document type	Journal Article ; Review
ZDB-ID	2388460-5
ISSN	1876-7958 ; 1673-5374
ISSN (online)	1876-7958
ISSN	1673-5374
DOI	10.4103/1673-5374.375318
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Article ; Online: Staining Methods for Normal and Regenerative Myelin in the Nervous System.

García-García, Óscar D / Weiss, Tamara / Chato-Astrain, Jesús / Raimondo, Stefania / Carriel, Víctor

Methods in molecular biology (Clifton, N.J.)

2022 Volume 2566, Page(s) 187–203

Abstract: Histochemical and fluorescence-based techniques enable the specific identification of myelin by bright-field or fluorescence microscopy. In this chapter, we describe four histological methods for the evaluation of myelin on peripheral nerve tissue ... ...

Abstract	Histochemical and fluorescence-based techniques enable the specific identification of myelin by bright-field or fluorescence microscopy. In this chapter, we describe four histological methods for the evaluation of myelin on peripheral nerve tissue sections. The first method combines the Luxol fast blue (LFB) technique with a modified Picrosirius staining contrasted with Harris hematoxylin, called MCOLL. This method simultaneously stains myelin, collagen fibers, and cell nuclei, thus giving an integrated overview of the histology, collagen network, and myelin content of the tissue in paraffin-embedded or cryosectioned samples. Secondly, we describe the osmium tetroxide method, which provides a permanent positive reaction for myelin as well as other lipids present in the tissue. The third method is the immunofluorescence-based detection of myelin proteins that allows to combine information about their expression status with other proteins of interest. Finally, the FluoroMyelin™ stains enable a fast detection of the myelin content that can be easily implemented in immunofluorescence staining panels for cryosectioned tissues. Together, this chapter provides a variety of methods to accurately identify myelin in different experimental approaches.
MeSH term(s)	Collagen/metabolism ; Coloring Agents/analysis ; Hematoxylin ; Lipids/analysis ; Myelin Sheath/metabolism ; Osmium Tetroxide ; Staining and Labeling
Chemical Substances	Coloring Agents ; Lipids ; Collagen (9007-34-5) ; Osmium Tetroxide (P40W033BGM) ; Hematoxylin (YKM8PY2Z55)
Language	English
Publishing date	2022-04-25
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ISSN	1940-6029
ISSN (online)	1940-6029
DOI	10.1007/978-1-0716-2675-7_15
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Article ; Online: Tissue Fixation and Processing for the Histological Identification of Lipids.

Sánchez-Porras, David / Bermejo-Casares, Fabiola / Carmona, Ramón / Weiss, Tamara / Campos, Fernando / Carriel, Víctor

Methods in molecular biology (Clifton, N.J.)

2022 Volume 2566, Page(s) 175–186

Abstract: Lipids are a heterogeneous group of substances characterized by their solubility in organic solvents and insolubility in water. Lipids can be found as normal components of different tissues and organs, and they can be affected by several pathological ... ...

Abstract	Lipids are a heterogeneous group of substances characterized by their solubility in organic solvents and insolubility in water. Lipids can be found as normal components of different tissues and organs, and they can be affected by several pathological conditions. The histochemical identification of lipids plays an important role in the histopathological diagnosis and research, but successful staining depends on adequate fixation and processing of the tissue. Here we describe methods to fix, cryoprotect, and process tissue samples for the histochemical identification of lipids in frozen or paraffin-embedded tissues.
MeSH term(s)	Formaldehyde ; Lipids ; Paraffin Embedding/methods ; Solvents ; Tissue Fixation/methods ; Water
Chemical Substances	Lipids ; Solvents ; Water (059QF0KO0R) ; Formaldehyde (1HG84L3525)
Language	English
Publishing date	2022-04-25
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ISSN	1940-6029
ISSN (online)	1940-6029
DOI	10.1007/978-1-0716-2675-7_14
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Article: Peripheral nerve regeneration through nerve conduits evokes differential expression of growth-associated protein-43 in the spinal cord.

Chato-Astrain, Jesús / Roda, Olga / Sánchez-Porras, David / Miralles, Esther / Alaminos, Miguel / Campos, Fernando / García-García, Óscar Darío / Carriel, Víctor

Neural regeneration research

2023 Volume 18, Issue 8, Page(s) 1852–1856

Abstract: Growth-associated protein 43 plays a key role in neurite outgrowth through cytoskeleton remodeling. We have previously demonstrated that structural damage of peripheral nerves induces growth-associated protein 43 upregulation to promote growth cone ... ...

Abstract	Growth-associated protein 43 plays a key role in neurite outgrowth through cytoskeleton remodeling. We have previously demonstrated that structural damage of peripheral nerves induces growth-associated protein 43 upregulation to promote growth cone formation. Conversely, the limited regenerative capacity of the central nervous system due to an inhibitory environment prevents major changes in neurite outgrowth and should be presumably associated with low levels of growth-associated protein 43 expression. However, central alterations due to peripheral nerve damage have never been assessed using the growth-associated protein 43 marker. In this study, we used the tubulization technique to repair 1 cm-long nerve gaps in the rat nerve injury/repair model and detected growth-associated protein 43 expression in the peripheral and central nervous systems. First, histological analysis of the regeneration process confirmed an active regeneration process of the nerve gaps through the conduit from 10 days onwards. The growth-associated protein 43 expression profile varied across regions and follow-up times, from a localized expression to an abundant and consistent expression throughout the regeneration tissue, confirming the presence of an active nerve regeneration process. Second, spinal cord changes were also histologically assessed, and no apparent changes in the structural and cellular organization were observed using routine staining methods. Surprisingly, remarkable differences and local changes appeared in growth-associated protein 43 expression at the spinal cord level, in particular at 20 days post-repair and beyond. Growth-associated protein 43 protein was first localized in the gracile fasciculus and was homogeneously distributed in the left posterior cord. These findings differed from the growth-associated protein 43 pattern observed in the healthy control, which did not express growth-associated protein 43 at these levels. Our results revealed a differential expression in growth-associated protein 43 protein not only in the regenerating nerve tissue but also in the spinal cord after peripheral nerve transection. These findings open the possibility of using this marker to monitor changes in the central nervous system after peripheral nerve injury.
Language	English
Publishing date	2023-01-11
Publishing country	India
Document type	Journal Article
ZDB-ID	2388460-5
ISSN	1876-7958 ; 1673-5374
ISSN (online)	1876-7958
ISSN	1673-5374
DOI	10.4103/1673-5374.363180
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Article ; Online: Histological characterization of the human scapholunate ligament.

Chato-Astrain, Jesús / Roda, Olga / Carriel, Víctor / Hita-Contreras, Fidel / Sánchez-Montesinos, Indalecio / Alaminos, Miguel / Hernández-Cortés, Pedro

Microscopy research and technique

2023 Volume 87, Issue 2, Page(s) 257–271

Abstract: The scapholunate interosseous ligament (SLIL) plays a fundamental role in stabilizing the wrist bones, and its disruption is a frequent cause of wrist arthrosis and disfunction. Traditionally, this structure is considered to be a variety of ... ...

Abstract	The scapholunate interosseous ligament (SLIL) plays a fundamental role in stabilizing the wrist bones, and its disruption is a frequent cause of wrist arthrosis and disfunction. Traditionally, this structure is considered to be a variety of fibrocartilaginous tissue and consists of three regions: dorsal, membranous and palmar. Despite its functional relevance, the exact composition of the human SLIL is not well understood. In the present work, we have analyzed the human SLIL and control tissues from the human hand using an array of histological, histochemical and immunohistochemical methods to characterize each region of this structure. Results reveal that the SLIL is heterogeneous, and each region can be subdivided in two zones that are histologically different to the other zones. Analysis of collagen and elastic fibers, and several proteoglycans, glycoproteins and glycosaminoglycans confirmed that the different regions can be subdivided in two zones that have their own structure and composition. In general, all parts of the SLIL resemble the histological structure of the control articular cartilage, especially the first part of the membranous region (zone M1). Cells showing a chondrocyte-like phenotype as determined by S100 were more abundant in M1, whereas the zone containing more CD73-positive stem cells was D2. These results confirm the heterogeneity of the human SLIL and could contribute to explain why certain zones of this structure are more prone to structural damage and why other zones have specific regeneration potential. RESEARCH HIGHLIGHTS: Application of an array of histological analysis methods allowed us to demonstrate that the human scapholunate ligament is heterogeneous and consists of at least six different regions sharing similarities with the human cartilage, ligament and other anatomical structures.
MeSH term(s)	Humans ; Wrist Joint ; Ligaments, Articular ; Cartilage, Articular ; Collagen ; Proteoglycans
Chemical Substances	Collagen (9007-34-5) ; Proteoglycans
Language	English
Publishing date	2023-09-28
Publishing country	United States
Document type	Journal Article
ZDB-ID	1099714-3
ISSN	1097-0029 ; 1059-910X
ISSN (online)	1097-0029
ISSN	1059-910X
DOI	10.1002/jemt.24428
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Article ; Online: Genipin crosslinking promotes biomechanical reinforcement and pro-regenerative macrophage polarization in bioartificial tubular substitutes.

Berasain, Jone / Ávila-Fernández, Paula / Cárdenas-Pérez, Rocío / Cànaves-Llabrés, Antoni Ignasi / Etayo-Escanilla, Miguel / Alaminos, Miguel / Carriel, Víctor / García-García, Óscar Darío / Chato-Astrain, Jesús / Campos, Fernando

Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

2024 Volume 174, Page(s) 116449

Abstract: Traumatic nerve injuries are nowadays a significant clinical challenge and new substitutes with adequate biological and mechanical properties are in need. In this context, fibrin-agarose hydrogels (FA) have shown the possibility to generate tubular ... ...

Abstract	Traumatic nerve injuries are nowadays a significant clinical challenge and new substitutes with adequate biological and mechanical properties are in need. In this context, fibrin-agarose hydrogels (FA) have shown the possibility to generate tubular scaffolds with promising results for nerve repair. However, to be clinically viable, these scaffolds need to possess enhanced mechanical properties. In this line, genipin (GP) crosslinking has demonstrated to improve biomechanical properties with good biological properties compared to other crosslinkers. In this study, we evaluated the impact of different GP concentrations (0.05, 0.1 and 0.2% (m/v)) and reaction times (6, 12, 24, 72 h) on bioartificial nerve substitutes (BNS) consisting of nanostructured FA scaffolds. First, crosslinked BNS were studied histologically, ultrastructurally and biomechanically and then, its biocompatibility and immunomodulatory effects were ex vivo assessed with a macrophage cell line. Results showed that GP was able to improve the biomechanical resistance of BNS, which were dependent on both the GP treatment time and concentration without altering the structure. Moreover, biocompatibility analyses on macrophages confirmed high cell viability and a minimal reduction of their metabolic activity by WST-1. In addition, GP-crosslinked BNS effectively directed macrophage polarization from a pro-inflammatory (M1) towards a pro-regenerative (M2) phenotype, which was in line with the cytokines release profile. In conclusion, this study considers time and dose-dependent effects of GP in FA substitutes which exhibited increased biomechanical properties while reducing immunogenicity and promoting pro-regenerative macrophage shift. These tubular substitutes could be useful for nerve application or even other tissue engineering applications such as urethra.
Language	English
Publishing date	2024-03-20
Publishing country	France
Document type	Journal Article
ZDB-ID	392415-4
ISSN	1950-6007 ; 0753-3322 ; 0300-0893
ISSN (online)	1950-6007
ISSN	0753-3322 ; 0300-0893
DOI	10.1016/j.biopha.2024.116449
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Article ; Online: A Novel In Vitro Pathological Model for Studying Neural Invasion in Non-Melanoma Skin Cancer.

Ávila-Fernández, Paula / Etayo-Escanilla, Miguel / Sánchez-Porras, David / Blanco-Elices, Cristina / Campos, Fernando / Carriel, Víctor / García-García, Óscar Darío / Chato-Astrain, Jesús

Gels (Basel, Switzerland)

2024 Volume 10, Issue 4

Abstract: Neural Invasion (NI) is a key pathological feature of cancer in the colonization of distant tissues, and its underlying biological mechanisms are still scarcely known. The complex interactions between nerve and tumor cells, along with the stroma, make it ...

Abstract	Neural Invasion (NI) is a key pathological feature of cancer in the colonization of distant tissues, and its underlying biological mechanisms are still scarcely known. The complex interactions between nerve and tumor cells, along with the stroma, make it difficult to reproduce this pathology in effective study models, which in turn has limited the understanding of NI pathogenesis. In this study, we have designed a three-dimensional model of NI squamous cell carcinoma combining human epidermoid carcinoma cells (hECCs) with a complete peripheral nerve segment encapsulated in a fibrine-agarose hydrogel. We recreated two vital processes of NI: a pre-invasive NI model in which hECCs were seeded on the top of the nerve-enriched stroma, and an invasive NI model in which cancer cells were immersed with the nerve in the hydrogel. Histological, histochemical and immunohistochemical analyses were performed to validate the model. Results showed that the integration of fibrin-agarose advanced hydrogel with a complete nerve structure and hECCs successfully generated an environment in which tumor cells and nerve components coexisted. Moreover, this model correctly preserved components of the neural extracellular matrix as well as allowing the proliferation and migration of cells embedded in hydrogel. All these results suggest the suitability of the model for the study of the mechanisms underlaying NI.
Language	English
Publishing date	2024-04-08
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2813982-3
ISSN	2310-2861 ; 2310-2861
ISSN (online)	2310-2861
ISSN	2310-2861
DOI	10.3390/gels10040252
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Article ; Online: Histochemical and Immunohistochemical Methods for the Identification of Proteoglycans.

Sánchez-Porras, David / Varas, Juan / Godoy-Guzmán, Carlos / Bermejo-Casares, Fabiola / San Martín, Sebastián / Carriel, Víctor

Methods in molecular biology (Clifton, N.J.)

2022 Volume 2566, Page(s) 85–98

Abstract: Proteoglycans (PGs) are non-fibrillar extracellular matrix (ECM) molecules composed by a protein core and glycosaminoglycan (GAG) chains. These molecules are present in all tissues playing essential structural, biomechanical, and biological roles. In ... ...

Abstract	Proteoglycans (PGs) are non-fibrillar extracellular matrix (ECM) molecules composed by a protein core and glycosaminoglycan (GAG) chains. These molecules are present in all tissues playing essential structural, biomechanical, and biological roles. In addition, PGs can regulate cell behavior due to their versatility and ability to interact with other ECM molecules, growth factors, and cells. The distribution of PGs can be evaluated by histochemical and immunohistochemical methods. Histochemical methods aimed to provide a useful overview of the presence and distribution pattern of certain groups of PGs. In contrast, immunohistochemical procedures aimed the identification of highly specific target molecules. In this chapter we described Alcian Blue, Safranin O, and Toluidine Blue histochemical methods for the screening of PGs in tissue sections. Finally, we describe the immunohistochemical procedures for specific identification of PGs (decorin, biglycan, and versican) in formaldehyde-fixed and paraffin-embedded tissues.
MeSH term(s)	Alcian Blue ; Biglycan ; Chondroitin Sulfate Proteoglycans/metabolism ; Decorin ; Extracellular Matrix Proteins/metabolism ; Formaldehyde ; Glycosaminoglycans/metabolism ; Tolonium Chloride ; Versicans
Chemical Substances	Biglycan ; Chondroitin Sulfate Proteoglycans ; Decorin ; Extracellular Matrix Proteins ; Glycosaminoglycans ; Versicans (126968-45-4) ; Tolonium Chloride (15XUH0X66N) ; Formaldehyde (1HG84L3525) ; Alcian Blue (P4448TJR7J)
Language	English
Publishing date	2022-04-25
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ISSN	1940-6029
ISSN (online)	1940-6029
DOI	10.1007/978-1-0716-2675-7_7
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Article ; Online: CURB-65 as a predictor of 30-day mortality in patients hospitalized with COVID-19 in Ecuador: COVID-EC study.

Carriel, J / Muñoz-Jaramillo, R / Bolaños-Ladinez, O / Heredia-Villacreses, F / Menéndez-Sanchón, J / Martin-Delgado, J

Revista clinica espanola

2021 Volume 222, Issue 1, Page(s) 37–41

Abstract: Objective: This article aims to assess the utility of CURB-65 in predicting 30-day mortality in adult patients hospitalized with COVID-19.: Methods: This work is a cohort study conducted between March 1 and April 30, 2020 in Ecuador.: Results: A ... ...

Abstract	Objective: This article aims to assess the utility of CURB-65 in predicting 30-day mortality in adult patients hospitalized with COVID-19. Methods: This work is a cohort study conducted between March 1 and April 30, 2020 in Ecuador. Results: A total of 247 patients were included (mean age 60 ± 14 years, 70% men, overall mortality 41.3%). Patients with CURB-65 ≥ 2 had a higher mortality rate (57 vs. 17%, p < .001) that was associated with other markers of risk: advanced age, hypertension, overweight/obesity, kidney failure, hypoxemia, requirement for mechanical ventilation, or onset of respiratory distress. Conclusions: CURB-65 ≥ 2 was associated with higher 30-day mortality on the univariate (Kaplan-Meier estimator) and multivariate (Cox regression) analysis.
MeSH term(s)	Adult ; Aged ; COVID-19 ; Cohort Studies ; Ecuador/epidemiology ; Female ; Humans ; Male ; Middle Aged ; Retrospective Studies ; SARS-CoV-2
Language	English
Publishing date	2021-04-30
Publishing country	Spain
Document type	Case Reports
ISSN	2254-8874
ISSN (online)	2254-8874
DOI	10.1016/j.rceng.2020.10.006
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Article: Comprehensive

García-García, Óscar Darío / El Soury, Marwa / Campos, Fernando / Sánchez-Porras, David / Geuna, Stefano / Alaminos, Miguel / Gambarotta, Giovanna / Chato-Astrain, Jesús / Raimondo, Stefania / Carriel, Víctor

Frontiers in bioengineering and biotechnology

2023 Volume 11, Page(s) 1162684

Abstract: As a reliable alternative to autografts, decellularized peripheral nerve allografts (DPNAs) should mimic the complex microstructure of native nerves and be immunogenically compatible. Nevertheless, there is a current lack of decellularization methods ... ...

Abstract	As a reliable alternative to autografts, decellularized peripheral nerve allografts (DPNAs) should mimic the complex microstructure of native nerves and be immunogenically compatible. Nevertheless, there is a current lack of decellularization methods able to remove peripheral nerve cells without significantly altering the nerve extracellular matrix (ECM). The aims of this study are firstly to characterize
Language	English
Publishing date	2023-03-30
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2719493-0
ISSN	2296-4185
ISSN	2296-4185
DOI	10.3389/fbioe.2023.1162684
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