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In Brazil, around 80% of snakebites are caused by snakes of the genus Bothrops. A three-dimensional culture model was standardized and used to perform treatments with Bothrops erythromelas venom (BeV) and its antivenom (AV). The MRC-5 and L929 cell lines were cultured at increasing cell densities. Morphometric parameters were evaluated through images obtained from an inverted microscope: solidity, circularity, and Feret diameter. L929 microtissues (MT) showed better morphometric data, and thus they were used for further analysis. MT viability was assessed using the acridine orange and ethidium bromide staining method, which showed viable cells in the MT on days 5, 7, and 10 of cultivation. Histochemical and histological analyses were performed, including hematoxylin/eosin staining, which showed a good structure of the spheroids. Alcian blue staining revealed the presence of acid proteoglycans. Immunohistochemical analysis with ki-67 showed different patterns of cell proliferation. The MT were also subjected to pharmacological tests using the BeV, in the presence or absence of its AV. The results showed that the venom was not cytotoxic, but it caused morphological changes. The MT showed cell detachment, losing their structure. The antivenom was able to partially prevent the venom activities.
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Hydrogels are polymeric biomaterials characterised by their promising biological and biomechanical properties, which make them potential alternatives for use in tendon repair. The aim of the present study was to generate in vitro, and determine the therapeutic efficacy in vivo, of novel nanostructured fibrin-based hydrogels to be used as an augmentation strategy for the surgical repair of rat Achilles tendon injuries. Fibrin, fibrin-agarose and fibrin-collagen nanostructured hydrogels (NFH, NFAH and NFCH, respectively) were generated and their biomechanical properties and cell-biomaterial interactions characterised ex vivo. Achilles tendon ruptures were created in 24 adult Wistar rats, which were next treated with direct repair (control group) or direct repair augmented with the generated biomaterials (6 rats/group). After 4 and 8 weeks, the animals were euthanised for macroscopical and histological analyses. Biomechanical characterisation showed optimal properties of the biomaterials for use in tendon repair. Moreover, biological analyses confirmed that tendon-derived fibroblasts were able to adhere to the surface of the generated biomaterials, with high levels of viability and functionality. In vivo studies demonstrated successful tendon repair in all groups. Lastly, histological analyses disclosed better tissue and extracellular matrix organisation and alignment with biomaterial-based augmentation strategies than direct repair, especially when NFAH and NFCH were used. The present study demonstrated that nanostructured fibrin-collagen hydrogels can be used to enhance the healing process in the surgical repair of tendon ruptures.
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Tendón Calcáneo , Traumatismos de los Tendones , Tendón Calcáneo/patología , Tendón Calcáneo/cirugía , Animales , Materiales Biocompatibles/farmacología , Colágeno/farmacología , Fibrina/farmacología , Hidrogeles/farmacología , Ratas , Ratas Wistar , Traumatismos de los Tendones/patología , Traumatismos de los Tendones/cirugíaRESUMEN
BACKGROUND: There is not an ideal biomaterial for tissue-engineered skin substitutes (TESSs), and most of the studies or existing therapies use xenogeneic origin natural biomaterials or biosynthetic scaffolds. OBJECTIVE: To analyse clinical, histological integration and homeostasis parameters of a human TESS manufactured with fibrin-hyaluronic acid biomaterial (HA-Skin), grafted in immunodeficient mice for 8 weeks, and compared with the gold standard treatment (Autograft), a human TESS manufactured with fibrin-agarose biomaterial (AG-Skin) and secondary wound healing dressings. METHODS: Human TESSs and autografts were implanted into BALB/c mice after surgical excision. Secondary wound healing approach was achieved with biosynthetic collagen wound dressing (Biobrane® ) and fibrin-hyaluronic acid or fibrin-agarose biomaterial without cells (Total N = 44). Clinical integration and homeostasis parameters were evaluated every two weeks for two months. Histological and immunohistochemical analyses were performed four and eight weeks after grafting. RESULTS: HA-Skin, AG-Skin and Autograft groups showed a proper clinical integration and epithelization eight weeks later. Scar evaluation revealed better results for Autograft and HA-Skin. Homeostasis analysis indicated similar values of transepidermal water loss and elasticity between HA-Skin (6.42 ± 0.75 g/h/m2 , 0.42 ± 0.08 AU), Autograft (6.91 ± 1.28 g/h/m2 , 0.40 ± 0.08 AU) and healthy mouse skin (6.40 ± 0.43 g/h/m2 , 0.35 ± 0.03 AU). Histological results showed that human TESSs and autografts presented better skin structuration and higher expression of cytokeratins. CONCLUSIONS: This study suggests that human TESS based on fibrin-hyaluronic acid biomaterial could be suitable for clinical application in the treatment of several dermatological pathologies (wound healing).
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Piel Artificial , Animales , Vendajes , Humanos , Ácido Hialurónico , Ratones , Ratones Endogámicos BALB C , Cicatrización de HeridasRESUMEN
The structure of the human skin is directly dependent on its location and the mechanical forces to which it is subjected. In the present work, we have performed a comprehensive analysis of the human ridged and non-ridged skin to identify the differences and similarities between both skin types. For this purpose, human skin samples were obtained from dorsal hand skin (DHS), palmar hand skin (PHS), dorsal foot skin (DFS) and plantar foot skin (PFS) from the same cadaveric donors. Histological, histochemical and semiquantitative and quantitative immunohistochemical analyses were carried out to evaluate the epidermis, dermis and basement membrane. Results show that the epithelial layer of ridged skin had larger cell number and size than non-ridged skin for most strata. Melanocytes and Langerhans cells were more abundant in non-ridged skin, whereas Merkel cells were preferentially found in ridged skin. The expression pattern of CK5/6 was slightly differed between non-ridged and ridged skin. Involucrin expression was slightly more intense in non-ridged skin than in ridged skin. Collagen was more abundant in foot skin dermis than in hand skin, and in ridged skin as compared to non-ridged skin. Elastic fibers were more abundant in DHS. Biglycan was more abundant in foot skin than in hand skin. No differences were found for blood and lymphatic vessels. The basement membrane laminin was preferentially found in foot skin. These results revealed important differences at the epithelial, dermal and basement membrane levels that could contribute to a better knowledge of the human skin histology.
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Piel/patología , Adulto , Anciano , Cadáver , Histocitoquímica , Humanos , Inmunohistoquímica , Persona de Mediana Edad , Donantes de TejidosRESUMEN
OBJECTIVES: Recently, the field of tissue engineering has made numerous advances towards achieving artificial tendon substitutes with excellent mechanical and histological properties, and has had some promising experimental results. The purpose of this systematic review is to assess the efficacy of tissue engineering in the treatment of tendon injuries. METHODS: We searched MEDLINE, Embase, and the Cochrane Library for the time period 1999 to 2016 for trials investigating tissue engineering used to improve tendon healing in animal models. The studies were screened for inclusion based on randomization, controls, and reported measurable outcomes. The RevMan software package was used for the meta-analysis. RESULTS: A total of 388 references were retrieved and 35 studies were included in this systematic review. The different biomaterials developed were analyzed and we found that they improve the biomechanical and histological characteristics of the repaired tendon. At meta-analysis, despite a high heterogeneity, it revealed a statistically significant effect in favour of the maximum load, the maximum stress, and the Young's modulus between experimental and control groups. In the forest plot, the diamond was on the right side of the vertical line and did not intersect with the line, favouring experimental groups. CONCLUSIONS: This review of the literature demonstrates the heterogeneity in the tendon tissue engineering literature. Several biomaterials have been developed and have been shown to enhance tendon healing and regeneration with improved outcomes.Cite this article: D. González-Quevedo, I. Martínez-Medina, A. Campos, F. Campos, V. Carriel. Tissue engineering strategies for the treatment of tendon injuries: a systematic review and meta-analysis of animal models. Bone Joint Res 2018;7:318-324. DOI: 10.1302/2046-3758.74.BJR-2017-0326.
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Microtissues (MT) are currently considered as a promising alternative for the fabrication of natural, 3D biomimetic functional units for the construction of bio-artificial substitutes by tissue engineering (TE). The aim of this study was to evaluate the possibility of generating mesenchymal cell-based MT using human umbilical cord Wharton's jelly stromal cells (WJSC-MT). MT were generated using agarose microchips and evaluated ex vivo during 28 days. Fibroblasts MT (FIB-MT) were used as control. Morphometry, cell viability and metabolism, MT-formation process and ECM synthesis were assessed by phase-contrast microscopy, functional biochemical assays, and histological analyses. Morphometry revealed a time-course compaction process in both MT, but WJSC-MT resulted to be larger than FIB-MT in all days analyzed. Cell viability and functionality evaluation demonstrated that both MT were composed by viable and metabolically active cells, especially the WJSC during 4-21 days ex vivo. Histology showed that WJSC acquired a peripheral pattern and synthesized an extracellular matrix-rich core over the time, what differed from the homogeneous pattern observed in FIB-MT. This study demonstrates the possibility of using WJSC to create MT containing viable and functional cells and abundant extracellular matrix. We hypothesize that WJSC-MT could be a promising alternative in TE protocols. However, future cell differentiation and in vivo studies are still needed to demonstrate the potential usefulness of WJSC-MT in regenerative medicine.
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Células Madre Mesenquimatosas/citología , Ingeniería de Tejidos/métodos , Gelatina de Wharton/citología , Supervivencia Celular , Matriz Extracelular/metabolismo , Humanos , Células Madre Mesenquimatosas/metabolismo , Gelatina de Wharton/metabolismoRESUMEN
Regeneration of the pulp-dentin complex with stem cells is a potential alternative to conventional root canal treatments. Human dental pulp stem cells (hDPSCs) have been extensively studied because of their ability to proliferate and differentiate into mineralized dental and non-dental tissues. Here we combined hDPSCs with two types of injectable poly-l-lactic acid (PLLA) microsphere with a nanofibrous or smooth surface to form bioactive injectable aggregates, and examined their ability to promote pulp regeneration in the root canal in an in vivo model. We investigated the biocompatibility, biosafety and odontogenic potential of fibrous (F-BIM) and smooth bioactive injectable microspheres (S-BIM) in vitro and in vivo. Our results demonstrated that PLLA microspheres and hDPSCs were able to form bioactive injectable aggregates that promoted dentin regeneration in both in vitro and in vivo models. Our results suggest that F-BIM and S-BIM may induce dentinogenesis upon in vivo grafting, and propose that the potential usefulness of the microsphere-hDPSC aggregates described here should be evaluated in clinical settings. Copyright © 2017 John Wiley & Sons, Ltd.
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Materiales Biocompatibles/farmacología , Pulpa Dental/citología , Endodoncia , Inyecciones , Microesferas , Nanofibras/química , Células Madre/citología , Investigación Biomédica Traslacional , Animales , Diferenciación Celular/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Forma de la Célula/efectos de los fármacos , Células Cultivadas , Dentina/metabolismo , Humanos , Macrófagos/efectos de los fármacos , Macrófagos/metabolismo , Masculino , Metaloproteinasa 14 de la Matriz/metabolismo , Ratones , Poliésteres/farmacología , Células Madre/efectos de los fármacosRESUMEN
Cell-derived matrices were recently described as novel biomaterials generated by human cells allowed to grow and synthetize their own extracellular matrix in culture. In the present work, we generated and evaluated a novel tissue-like substitute (WDM) consisting of a membrane derived from cultured human Wharton's jelly stem cells. WDM were evaluated ex vivo and in vivo by histochemistry and immunohistochemistry for several mesenchymal cell markers and fibrillar and non-fibrillar extracellular matrix components. Results show that WDM were heterogeneous and consisted of dense cell-poor areas surrounded by cell-rich zones with abundant HWJSC. Histological analyses demonstrated that cell-poor areas were very rich in fibrillar and non-fibrillar extracellular matrix components such as collagen and proteoglycans, and cells in the WDM were highly viable and mostly PCNA-positive. HWJSC in the WDM expressed all markers of this cell type, including CD90, CD105, pan cytokeratin and CK8. In vivo analysis showed that the WDM was highly biocompatible and grafting this membrane in the muscle of laboratory rats was not associated to increased inflammation, necrosis, tumorigenesis or other side effects, while cells properly integrated at the damage site and showed high proliferation index. These results suggest that the structure and composition of the extracellular matrix of these novel WDM could reproduce the situation of native human tissues and that WDM implanted in vivo are highly biocompatible and rapidly integrate in the host tissues. For these reasons, we hypothesize that WDM could be used in regenerative medicine protocols.
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Matriz Extracelular , Células Madre Mesenquimatosas/citología , Ingeniería de Tejidos/métodos , Gelatina de Wharton/citología , Animales , Células Cultivadas , Xenoinjertos , Humanos , Masculino , Membranas , Ratas , Ratas Wistar , Cordón Umbilical/citologíaRESUMEN
We analyze the effect of nanoparticle concentration on the physical properties of magnetic hydrogels consisting of polymer networks of the human fibrin biopolymer with embedded magnetic particles, swollen by a water-based solution. We prepared these magnetic hydrogels by polymerization of mixtures consisting mainly of human plasma and magnetic nanoparticles with OH- functionalization. Microscopic observations revealed that magnetic hydrogels presented some cluster-like knots that were connected by several fibrin threads. By contrast, nonmagnetic hydrogels presented a homogeneous net-like structure with only individual connections between pairs of fibers. The rheological analysis demonstrated that the rigidity modulus, as well as the viscoelastic moduli, increased quadratically with nanoparticle content following a square-like function. Furthermore, we found that time for gel point was shorter in the presence of magnetic nanoparticles. Thus, we can conclude that nanoparticles favor the cross-linking process, serving as nucleation sites for the attachment of the fibrin polymer. Attraction between the positive groups of the fibrinogen, from which the fibrin is polymerized, and the negative OH- groups of the magnetic particle surface qualitatively justifies the positive role of the nanoparticles in the enhancement of the mechanical properties of the magnetic hydrogels. Indeed, we developed a theoretical model that semiquantitatively explains the experimental results by assuming the indirect attraction of the fibrinogen through the attached nanoparticles. Due to this attraction the monomers condense into nuclei of the dense phase and by the end of the polymerization process the nuclei (knots) of the dense phase cross-link the fibrin threads, which enhances their mechanical properties.
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Materiales Biocompatibles/química , Hidrogeles/química , Imanes/química , Fenómenos Mecánicos , Nanopartículas/química , Humanos , Reología , Resistencia al Corte , Estrés MecánicoRESUMEN
INTRODUCTION: Human umbilical cord stem cells have inherent differentiation capabilities and potential usefulness in regenerative medicine. However, the epithelial differentiation capability and the heterogeneity of these cells have not been fully explored to the date. METHODS: We analyzed the expression of several undifferentiation and epithelial markers in cells located in situ in different zones of the umbilical cord -in situ analysis- and in primary ex vivo cell cultures of Wharton's jelly stem cells by microarray and immunofluorescence. RESULTS: Our results demonstrated that umbilical cord cells were heterogeneous and had intrinsic capability to express in situ stem cell markers, CD90 and CD105 and the epithelial markers cytokeratins 3, 4, 7, 8, 12, 13, 19, desmoplakin and zonula occludens 1 as determined by microarray and immunofluorescence, and most of these markers remained expressed after transferring the cells from the in situ to the ex vivo cell culture conditions. However, important differences were detected among some cell types in the umbilical cord, with subvascular zone cells showing less expression of stem cell markers and cells in Wharton's jelly and the amnioblastic zones showing the highest expression of stem cells and epithelial markers. CONCLUSIONS: These results suggest that umbilical cord mesenchymal cells have intrinsic potential to express relevant epithelial markers, and support the idea that they could be used as alternative cell sources for epithelial tissue engineering.
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Diferenciación Celular , Células Madre Mesenquimatosas/citología , Células Madre/citología , Cordón Umbilical/citología , Gelatina de Wharton/citología , Antígenos CD/metabolismo , Biomarcadores/metabolismo , Células Cultivadas , Endoglina , Humanos , Queratinas/metabolismo , Células Madre Mesenquimatosas/metabolismo , Receptores de Superficie Celular/metabolismo , Células Madre/metabolismo , Antígenos Thy-1/metabolismo , Cordón Umbilical/metabolismo , Gelatina de Wharton/metabolismoRESUMEN
El vitiligo es una enfermedad pigmentaria cutánea adquirida e idiopática, debido a múltiples factores causales que ocasionan la destrucción o inactivación funcional del melanocito. Ha habido numerosas alternativas terapéuticas a lo largo de los años como: tratamientos tópicos, PUVA (psoralenos más luz ultravioleta) o fototerapia UVB de banda estrecha; sin embargo, muchos no responden a estos tratamientos médicos. El tratamiento quirúrgico del vitiligo se indica cuando las máculas hipo o amelanóticas, se localizan en zonas mal respondedoras o tras fracaso de los tratamientos convencionales. Se trata de favorecer la re-pigmentación de las lesiones, a través del trasplante de melanocitos desde las áreas pigmentadas normales. El objetivo de esta revisión, es el de analizar el estado actual del tratamiento quirúrgico del vitiligo y en concreto, del trasplante de melanocitos autólogos.
Vitiligo is an idiopathic and acquired disorder that affects the pigmentation of the skin due to several factors that cause the destruction of melanocytes. There are numerous therapeutic alternatives, topical treatments, PUVA or narrowband UVB phototherapy. However, many patients do not respond to these medical treatments. Surgical treatment of vitiligo is indicated when depigmented macules are located in poorly responsive or after failure of conventional treatments. It seeks to encourage repigmentation of lesions by transplantation of melanocytes from normal pigmented areas. The objective of this review is to analyze surgical treatment of vitiligo and especially autologous melanocyte transplantation.
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El vitiligo es una enfermedad pigmentaria cutánea adquirida e idiopática, debido a múltiples factores causales que ocasionan la destrucción o inactivación funcional del melanocito. Ha habido numerosas alternativas terapéuticas a lo largo de los años como: tratamientos tópicos, PUVA (psoralenos más luz ultravioleta) o fototerapia UVB de banda estrecha; sin embargo, muchos no responden a estos tratamientos médicos. El tratamiento quirúrgico del vitiligo se indica cuando las máculas hipo o amelanóticas, se localizan en zonas mal respondedoras o tras fracaso de los tratamientos convencionales. Se trata de favorecer la re-pigmentación de las lesiones, a través del trasplante de melanocitos desde las áreas pigmentadas normales. El objetivo de esta revisión, es el de analizar el estado actual del tratamiento quirúrgico del vitiligo y en concreto, del trasplante de melanocitos autólogos.(AU)
Vitiligo is an idiopathic and acquired disorder that affects the pigmentation of the skin due to several factors that cause the destruction of melanocytes. There are numerous therapeutic alternatives, topical treatments, PUVA or narrowband UVB phototherapy. However, many patients do not respond to these medical treatments. Surgical treatment of vitiligo is indicated when depigmented macules are located in poorly responsive or after failure of conventional treatments. It seeks to encourage repigmentation of lesions by transplantation of melanocytes from normal pigmented areas. The objective of this review is to analyze surgical treatment of vitiligo and especially autologous melanocyte transplantation.(AU)
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El folículo piloso-sebáceo o unidad folículo-sebácea-apócrina es un órgano microscópico complejo funcional y estructuralmente. Consta de un segmento superior, estable y un segmento inferior, que se regenera en cada ciclo piloso. El folículo piloso contiene una gran diversidad de tipos celulares, entre los que se encuentran células madre. De hecho, supone el nicho más importante de células madre en la piel, con la ventaja añadida de su fácil accesibilidad. Estas células madre se localizan en una prominencia a nivel de la inserción del músculo erector del pelo, llamada "bulge" (promontorio). Son las encargadas de regenerar el folículo en cada ciclo y también intervienen en la reconstitución de las glándulas sebáceas y de la epidermis interfolicular, en caso de lesión epidérmica. El conocimiento de la estructura del folículo piloso, está haciendo posible cada vez más el empleo de los distintos tipos celulares, especialmente, las células madre, en la ingeniería tisular de la piel. Asimismo, estudios recientes han diferenciado las células madres foliculares a diferentes estirpes, como por ejemplo: células nerviosas, hematopoyéticas y vasculares. Además, se ha estudiado la construcción de folículos pilosos, con resultados satisfactorios en ratones, aunque no del todo superponibles a humanos, en los que se precisan más investigaciones. La posibilidad de generar folículos pilosos humanos, supondría una revolución en el amplio campo de las alopecias. El objetivo de esta revisión es describir la estructura anatómica e histológica del folículo piloso, enfatizando en la importancia del mismo como nicho de células madre y su potencial utilidad en el campo de la ingeniería tisular, para la construcción de diversos tipos de tejidos.
The hair follicle is a microscopic organ, functionally and structurally complex. It can be divided into two distinct segments, the upper portion, stable and the lower portion, that undergoes regeneration every hair cycle. The hair follicle contains a variety of cells types, including stem cells. It fact, it is the most important niche of stem cells in the skin, with the added advantage of its easy accessibility. These stem cells are located in a prominence at the level of the insertion of the arrector pili muscle, called bulge. They are responsible of hair follicle regeneration each hair cycle, and also can form sebaceous glands and help in repopulation of the interfollicular epidermis after injury. The knowledge of hair follicle structure is making possible the use of different types of cells, especially stem cells, in skin tissue engineering. Furthermore, recent studies have differentiated follicle stem cells into different strains, for example: nervous, hematopoietic and vascular cells. Moreover, other late studies have focused on the hair follicle construction, with satisfactory results in mice, but not completely transferable to humans, which further research is needed. The possibility of human hair follicle regeneration would suppose a revolution in the broad world of alopecia. The aim of this review is to describe the anatomical and histological structure of the hair follicle, emphasizing the importance as stem cell niche, and its potential usefulness in the field of tissue engineering for the construction of various types of tissues.
RESUMEN
El folículo piloso-sebáceo o unidad folículo-sebácea-apócrina es un órgano microscópico complejo funcional y estructuralmente. Consta de un segmento superior, estable y un segmento inferior, que se regenera en cada ciclo piloso. El folículo piloso contiene una gran diversidad de tipos celulares, entre los que se encuentran células madre. De hecho, supone el nicho más importante de células madre en la piel, con la ventaja añadida de su fácil accesibilidad. Estas células madre se localizan en una prominencia a nivel de la inserción del músculo erector del pelo, llamada "bulge" (promontorio). Son las encargadas de regenerar el folículo en cada ciclo y también intervienen en la reconstitución de las glándulas sebáceas y de la epidermis interfolicular, en caso de lesión epidérmica. El conocimiento de la estructura del folículo piloso, está haciendo posible cada vez más el empleo de los distintos tipos celulares, especialmente, las células madre, en la ingeniería tisular de la piel. Asimismo, estudios recientes han diferenciado las células madres foliculares a diferentes estirpes, como por ejemplo: células nerviosas, hematopoyéticas y vasculares. Además, se ha estudiado la construcción de folículos pilosos, con resultados satisfactorios en ratones, aunque no del todo superponibles a humanos, en los que se precisan más investigaciones. La posibilidad de generar folículos pilosos humanos, supondría una revolución en el amplio campo de las alopecias. El objetivo de esta revisión es describir la estructura anatómica e histológica del folículo piloso, enfatizando en la importancia del mismo como nicho de células madre y su potencial utilidad en el campo de la ingeniería tisular, para la construcción de diversos tipos de tejidos.(AU)
The hair follicle is a microscopic organ, functionally and structurally complex. It can be divided into two distinct segments, the upper portion, stable and the lower portion, that undergoes regeneration every hair cycle. The hair follicle contains a variety of cells types, including stem cells. It fact, it is the most important niche of stem cells in the skin, with the added advantage of its easy accessibility. These stem cells are located in a prominence at the level of the insertion of the arrector pili muscle, called bulge. They are responsible of hair follicle regeneration each hair cycle, and also can form sebaceous glands and help in repopulation of the interfollicular epidermis after injury. The knowledge of hair follicle structure is making possible the use of different types of cells, especially stem cells, in skin tissue engineering. Furthermore, recent studies have differentiated follicle stem cells into different strains, for example: nervous, hematopoietic and vascular cells. Moreover, other late studies have focused on the hair follicle construction, with satisfactory results in mice, but not completely transferable to humans, which further research is needed. The possibility of human hair follicle regeneration would suppose a revolution in the broad world of alopecia. The aim of this review is to describe the anatomical and histological structure of the hair follicle, emphasizing the importance as stem cell niche, and its potential usefulness in the field of tissue engineering for the construction of various types of tissues.(AU)
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We contribute the first immunohistochemical study of MT1 melatonin receptor in Warthin's tumor and normal parotid gland. All 14 Warthin's tumors studied showed intense cytoplasmic positivity for MT1 receptor in all cylindrical epithelial cells lining spaces and a less intense positivity in basal cells. The lymphoid component accompanying the tumor was always negative for MT1 receptor. The parotid structure surrounding the tumor showed intense cytoplasmic positivity in all cells lining excretory ducts (lobar and lobulillar), with a lesser and focal positivity in cells of the acinar component. The biological activity of MT1 receptor in epithelial cells lining parotid excretory ducts may resemble its activity in Warthin's tumor cells. Hence, we propose Warthin's tumor as a useful positive control in immunohistochemical studies of MT1 melatonin receptor.
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Adenolinfoma/metabolismo , Receptor de Melatonina MT1/biosíntesis , Adenolinfoma/genética , Adenolinfoma/patología , Adulto , Células Epiteliales/metabolismo , Células Epiteliales/patología , Femenino , Humanos , Inmunohistoquímica , Masculino , Persona de Mediana Edad , Glándula Parótida/metabolismo , Glándula Parótida/patología , Receptor de Melatonina MT1/genética , Receptor de Melatonina MT1/metabolismoRESUMEN
INTRODUCTION: Smoothelin is a cytoskeletal protein of differentiated smooth muscle cells with contractile capacity, distinguishing it from other smooth muscle proteins, such as smooth muscle actin (SMA). OBJECTIVE: To evaluate the expression of smoothelin and SMA in the skin in order to establish specific localizations of smoothelin in smooth muscle cells with high contractile capacity and in the epithelial component of cutaneous adnexal structures. METHODS: Immunohistochemical analysis (smoothelin and SMA) was performed in 18 patients with normal skin. RESULTS: SMA was expressed by the vascular structures of superficial, deep, intermediate and adventitial plexuses, whereas smoothelin was specifically expressed in the cytoplasm of smooth muscle cells of the deepest vascular plexus and in no other plexus of the dermis. The hair erector muscle showed intense expression of smoothelin and SMA. Cells with nuclear expression of smoothelin and cytoplasmic expression of SMA were observed in the outer root sheath of the inferior portion of the hair follicles and intense cytoplasmic expression in cells of the dermal sheath to SMA. CONCLUSIONS: We report the first study of smoothelin expression in normal skin, which differentiates the superficial vascular plexus from the deep. The deep plexus comprises vessels with high contractile capacity, which is important for understanding dermal hemodynamics in normal skin and pathological processes. We suggest that the function of smoothelin in the outer root sheath may be to enhance the function of SMA, which has been related to mechanical stress. Smoothelin has not been studied in cutaneous pathology; however we believe it may be a marker specific for the diagnosis of leiomyomas and leiomyosarcomas of the skin. Also, smoothelin could differentiate arteriovenous malformations of cavernous hemangioma of the skin.
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Actinas/biosíntesis , Proteínas del Citoesqueleto/biosíntesis , Folículo Piloso/metabolismo , Proteínas Musculares/biosíntesis , Músculo Liso/metabolismo , Piel/metabolismo , Adulto , Femenino , Humanos , Inmunohistoquímica , Masculino , Persona de Mediana Edad , Piel/irrigación sanguíneaRESUMEN
During embryo implantation, invasive trophoblast cells mediate embryo invasion into the decidualized stroma, forming a rich network of lacunae that connect the embryonic tissues to the maternal blood vessels. Placentation is probably guided by the composition and organization of the endometrial extracellular matrix. Certain pathological conditions that occur during pregnancy, including diabetes, have been linked to abnormal placental morphology and consequent fetal morbidity. We used immunoperoxidase techniques to identify members of the collagen, proteoglycan and glycoprotein families in the various compartments of the rat placenta and to determine whether experimentally induced diabetes affects placental morphology and alters the distribution of these molecules during pregnancy. Single injections of alloxan (40 mg kg(-1) i.v.) were used to induce diabetes on day 2 of pregnancy in Wistar rats. Placentas were collected on days 14, 17, and 20. Type I and III collagen, as well as the proteoglycans decorin and biglycan, were found to be distributed throughout the placentas of control and diabetic rats. In both groups, laminin expression decreased at the end of pregnancy. In contrast, fibronectin was detected in the labyrinth region of diabetic rats at all gestational stages studied, whereas it was detected only at term pregnancy in the placentas of control rats. These results show for the first time that some extracellular matrix molecules are modulated during placental development. However, as diabetic rats presented increased fibronectin deposition exclusively in the labyrinth region, we speculate that diabetes alters the microenvironment at the maternal-fetal interface, leading to developmental abnormalities in the offspring.