16 research outputs found

    Alginate-Agarose Hydrogels Improve the In Vitro Differentiation of Human Dental Pulp Stem Cells in Chondrocytes. A Histological Study

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    [EN] Matrix-assisted autologous chondrocyte implantation (MACI) has shown promising results for cartilage repair, combining cultured chondrocytes and hydrogels, including alginate. The ability of chondrocytes for MACI is limited by different factors including donor site morbidity, dedifferentiation, limited lifespan or poor proliferation in vitro. Mesenchymal stem cells could represent an alternative for cartilage regeneration. In this study, we propose a MACI scaffold consisting of a mixed alginate-agarose hydrogel in combination with human dental pulp stem cells (hDPSCs), suitable for cartilage regeneration. Scaffolds were characterized according to their rheological properties, and their histomorphometric and molecular biology results. Agarose significantly improved the biomechanical behavior of the alginate scaffolds. Large scaffolds were manufactured, and a homogeneous distribution of cells was observed within them. Although primary chondrocytes showed a greater capacity for chondrogenic differentiation, hDPSCs cultured in the scaffolds formed large aggregates of cells, acquired a rounded morphology and expressed high amounts of type II collagen and aggrecan. Cells cultured in the scaffolds expressed not only chondral matrix-related genes, but also remodeling proteins and chondrocyte differentiation factors. The degree of differentiation of cells was proportional to the number and size of the cell aggregates that were formed in the hydrogels.This work was funded by the Ministry of Economy and Competitiveness of the Spanish Government (PID2019-106099RB-C42, MM) and by the Generalitat Valenciana, Spain (PROMETEO/2020/069, CC). CIBER-BBN and CIBER-ER are financed by the VI National R&D&I Plan 2008-2011, Iniciativa Ingenio 2010, Consolider Program, CIBER Actions and the Instituto de Salud Carlos III, with assistance of the European Regional Development Fund.Oliver-Ferrándiz, M.; Milián, L.; Sancho-Tello, M.; Martín De Llano, JJ.; Gisbert-Roca, F.; Martínez-Ramos, C.; Carda, C.... (2021). Alginate-Agarose Hydrogels Improve the In Vitro Differentiation of Human Dental Pulp Stem Cells in Chondrocytes. A Histological Study. Biomedicines. 9(7):1-22. https://doi.org/10.3390/biomedicines9070834S1229

    A standardised approach to the biomechanical evaluation of tracheal grafts

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    [EN] The ideal tracheal substitute must have biomechanical properties comparable to the native trachea, but currently there is no standardised approach to evaluating these properties. Here we propose a novel method for evaluating and comparing the properties of tracheal substitutes, thus systematising both measurement and data curation. This system was tested by comparing native rabbit tracheas to frozen and decellularised specimens and determining the histological characteristics of those specimens. We performed radial compression tests on the anteroposterior tracheal axis and longitudinal axial tensile tests with the specimens anastomosed to the jaw connected to a measuring system. All calculations and results were adjusted according to tracheal size, always using variables relative to the tracheal dimensions, thus permitting comparison of different sized organs. The biomechanical properties of the decellularised specimens were only slightly reduced compared to controls and significant in regard to the maximum stress withstood in the longitudinal axis (-0.246 MPa CI [-0.248, -0.145] MPa) and the energy stored per volume unit (-0.124 mJ & BULL;mm(-3) CI [-0.195, -0.055] mJ & BULL;mm(-3)). The proposed method is suitable for the systematic characterisation of the biomechanical properties of different tracheal substitutes, regardless of the size or nature of the substitute, thus allowing for direct comparisons.This research was funded by the 2018 Spanish Society of Thoracic Surgery grant to National Multicentric Study [Number 180101 to N.J.M.-H.] and [PI16-01315 to M.M.-R.] from the Instituto de Salud Carlos III. CIBERER is funded by the VI National R&D&I Plan 2008-2011, Iniciativa Ingenio 2010, Consolider Program, CIBER Actions and the Instituto de Salud Carlos III, with assistance from the European Regional Development Fund.Martínez-Hernández, NJ.; Más Estellés, J.; Milián-Medina, L.; Martínez-Ramos, C.; Cerón-Navarro, J.; Galbis-Caravajal, J.; Roig-Bataller, A.... (2021). A standardised approach to the biomechanical evaluation of tracheal grafts. Biomolecules. 11(10):1-12. https://doi.org/10.3390/biom11101461S112111

    Optimization of a decellularized protocol of porcine tracheas. Long-term effects of cryopreservation. A histological study

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    [EN] Objective: The aim of this study was to optimize a decellularization protocol in the trachea of Sus scrofa domestica (pig) as well as to study the effects of long-term cryopreservation on the extracellular matrix of decellularized tracheas. Methods: Porcine tracheas were decellularized using Triton X-100, SDC, and SDS alone or in combination. The effect of these detergents on the extracellular matrix characteristics of decellularized porcine tracheas was evaluated at the histological, biomechanical, and biocompatibility level. Morphometric approaches were used to estimate the effect of detergents on the collagen and elastic fibers content as well as on the removal of chondrocytes from decellularized organs. Moreover, the long-term structural, ultrastructural, and biomechanical effect of cryopreservation of decellularized tracheas were also estimated. Results: Two percent SDS was the most effective detergent tested concerning cell removal and preservation of the histological and biomechanical properties of the tracheal wall. However, long-term cryopreservation had no an appreciable effect on the structure, ultrastructure, and biomechanics of decellularized tracheal rings. Conclusion: The results presented here reinforce the use of SDS as a valuable decellularizing agent for porcine tracheas. Furthermore, a cryogenic preservation protocol is described, which has minimal impact on the histological and biomechanical properties of decellularized porcine tracheas.The author(s) disclosed receipt of the following financial support for the research, authorship, and/or publication of this article: This work was supported by grants MAT2016-76039-C4-2-R (MST) and PID2019-106099RB-C42 (MM) from the Ministry of Economy and Competitiveness of the Spanish Government, by grant PI16-01315 from the ISCIII (Ministerio de Ciencia, Innovacion y Universidades, Spain), and by grant PROMETEO/2020/069 (CC) from the local government of the Comunitat Valenciana (Spain), CIBER-BBN and CIBERER are funded by the VI National R&D&I Plan 2008-2011, Iniciativa Ingenio 2010, Consolider Program, CIBER Actions, and the Instituto de Salud Carlos III, with assistance from the European Regional Development Fund.Milián, L.; Sancho-Tello, M.; Roig-Soriano, J.; Foschini, G.; Martínez-Hernández, NJ.; Más Estellés, J.; Ruiz-Sauri, A.... (2021). Optimization of a decellularized protocol of porcine tracheas. Long-term effects of cryopreservation. A histological study. The International Journal of Artificial Organs. 44(12):998-1012. https://doi.org/10.1177/03913988211008912S9981012441

    A cell-free approach with a supporting biomaterial in the form of dispersed microspheres induces hyaline cartilage formation in a rabbit knee model

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    [EN] The objective of this study was to test a regenerative medicine strategy for the regeneration of articular cartilage. This approach combines microfracture of the subchondral bone with the implant at the site of the cartilage defect of a supporting biomaterial in the form of microspheres aimed at creating an adequate biomechanical environment for the differentiation of the mesenchymal stem cells that migrate from the bone marrow. The possible inflammatory response to these biomaterials was previously studied by means of the culture of RAW264.7 macrophages. The microspheres were implanted in a 3¿mm-diameter defect in the trochlea of the femoral condyle of New Zealand rabbits, covering them with a poly(l-lactic acid) (PLLA) membrane manufactured by electrospinning. Experimental groups included a group where exclusively PLLA microspheres were implanted, another group where a mixture of 50/50 microspheres of PLLA (hydrophobic and rigid) and others of chitosan (a hydrogel) were used, and a third group used as a control where no material was used and only the membrane was covering the defect. The histological characteristics of the regenerated tissue have been evaluated 3 months after the operation. We found that during the regeneration process the microspheres, and the membrane covering them, are displaced by the neoformed tissue in the regeneration space toward the subchondral bone region, leaving room for the formation of a tissue with the characteristics of hyaline cartilage.Comisión de Investigaciones Científicas de la Provincia de Buenos Aires (CICPBA), Universidad Nacional de La Plata, Grant/Award Number: 11/X643; Agencia Estatal de Investigación/Fondo Europeo de Desarrollo Regional de la Unión Europea, Grant/Award Number: MAT2016-76039-C4-1 2-R; Spanish Ministry of Economy and Competitiveness (MINECO)Zurriaga Carda, J.; Lastra, ML.; Antolinos-Turpin, CM.; Morales-Román, RM.; Sancho-Tello, M.; Perea-Ruiz, S.; Milián, L.... (2020). A cell-free approach with a supporting biomaterial in the form of dispersed microspheres induces hyaline cartilage formation in a rabbit knee model. 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H., & Hui, J. H. P. (2013). Evidence-Based Status of Microfracture Technique: A Systematic Review of Level I and II Studies. Arthroscopy: The Journal of Arthroscopic & Related Surgery, 29(9), 1579-1588. doi:10.1016/j.arthro.2013.05.027Hangody, L., Kish, G., Kárpáti, Z., Udvarhelyi, I., Szigeti, I., & Bély, M. (1998). Mosaicplasty for the Treatment of Articular Cartilage Defects: Application in Clinical Practice. Orthopedics, 21(7), 751-756. doi:10.3928/0147-7447-19980701-04Hoemann, C., Kandel, R., Roberts, S., Saris, D. B. F., Creemers, L., Mainil-Varlet, P., … Buschmann, M. D. (2011). International Cartilage Repair Society (ICRS) Recommended Guidelines for Histological Endpoints for Cartilage Repair Studies in Animal Models and Clinical Trials. CARTILAGE, 2(2), 153-172. doi:10.1177/1947603510397535Kumar, M. N. V. R., Muzzarelli, R. A. A., Muzzarelli, C., Sashiwa, H., & Domb, A. J. (2004). Chitosan Chemistry and Pharmaceutical Perspectives. 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Journal of Biomedical Materials Research Part A, 106(2), 570-579. doi:10.1002/jbm.a.36260Lastra, M. L., Molinuevo, M. S., Cortizo, A. M., & Cortizo, M. S. (2016). Fumarate Copolymer-Chitosan Cross-Linked Scaffold Directed to Osteochondrogenic Tissue Engineering. Macromolecular Bioscience, 17(5). doi:10.1002/mabi.201600219Lebourg, M., Martínez-Díaz, S., García-Giralt, N., Torres-Claramunt, R., Ribelles, J. G., Vila-Canet, G., & Monllau, J. (2013). Cell-free cartilage engineering approach using hyaluronic acid–polycaprolactone scaffolds: A study in vivo. Journal of Biomaterials Applications, 28(9), 1304-1315. doi:10.1177/0885328213507298Luzardo-Alvarez, A., Blarer, N., Peter, K., Romero, J. F., Reymond, C., Corradin, G., & Gander, B. (2005). Biodegradable microspheres alone do not stimulate murine macrophages in vitro, but prolong antigen presentation by macrophages in vitro and stimulate a solid immune response in mice. 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Der Orthopäde, 28(1), 26-32. doi:10.1007/pl00003545Tetè, S., Mastrangelo, F., Carone, L., Nargi, E., Costanzo, G., Vinci, R., … Ciccarelli, R. (2007). Morphostructural Analysis of Human Follicular Stem Cells on Highly Porous Bone Hydroxyapatite Scaffold. International Journal of Immunopathology and Pharmacology, 20(4), 819-826. doi:10.1177/039463200702000418Van den Borne, M. P. J., Raijmakers, N. J. H., Vanlauwe, J., Victor, J., de Jong, S. N., Bellemans, J., & Saris, D. B. F. (2007). International Cartilage Repair Society (ICRS) and Oswestry macroscopic cartilage evaluation scores validated for use in Autologous Chondrocyte Implantation (ACI) and microfracture. Osteoarthritis and Cartilage, 15(12), 1397-1402. doi:10.1016/j.joca.2007.05.005Vikingsson, L., Sancho-Tello, M., Ruiz-Saurí, A., Díaz, S. M., Gómez-Tejedor, J. A., Ferrer, G. G., … Ribelles, J. L. G. (2015). 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    EURIN: Rea's Rescue. Modelado 3D y animación de personajes y programación para un nivel del videojuego

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    [ES] 2 El objeto de este trabajo de fin de grado es mi participación en el desarrollo del prototipo un nivel tutorial de un videojuego de plataformas 2D: Eurin, Rea’s rescue. Este prototipo se ha trabajado por un grupo de cuatro estudiantes de Bellas Artes. En este videojuego basado en la mitología griega, el jugador, encarnado en Eurin -una diosa primigénia- deberá explorar el Tártaro evitando los obstáculos que se le interpongan y consiguiendo los objetivos que le permitan escapar de él.El reducido tamaño del equipo conllevó que me hiciese cargo de más de un rol en el desarrollo del juego. Mientras que la redacción del diseño de jugabilidad se realizó de forma conjunta, el resto de áreas se dividieron. Así pues, yo me encargué de las labores de desarrollo 3D del personaje principal (en las que se incluyen: modelado,rigging y skinning), la implementación de todos los componentes en el motor gráfico y la programaciónMilián Lacomba, L. (2015). EURIN: Rea's Rescue. Modelado 3D y animación de personajes y programación para un nivel del videojuego. http://hdl.handle.net/10251/57355.TFG

    Estrategia para lograr un uso racional de los medicamentos

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    Uno de los objetivos del Programa Nacional de Medicamentos del Ministerio de Salud Pública cubano es que se haga un buen uso de los medicamentos, pero para ello se requiere de la participación consciente y activa de diferentes eslabones como el productor, el prescriptor, el dispensador, y por último, el consumidor, que constituye el determinante final de su uso. Esta estrategia es una vía para alcanzar este objetivo mediante la participación comunitaria y la intersectorialidad.<br>One of the objectives of the National Program of Drugs of the Ministry of Public Health is the proper use of drugs. However, for attaining this, the conscious and active involvement of different links such as the manufacturer, the physician, the seller and finally the consumer, who determines the ultimate use of the drug, is required. This strategy is a way for achieving this goal on the basis of the community participation and inter-sector involvement

    Results of the application of Heberprot-P for the treatment of diabetic foot ulcer

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    Foundation: diabetes mellitus is a chronic disease. One of its frequent complications is the diabetic foot. For its treatment Heberprot-P is used.Objective: to describe the results of the application of Heberprot-P for the treatment of diabetic foot ulcer. Methods: prospective descriptive study conducted in the Diabetic Care Consultation, during the period from April 2009 to April 2016. The sample consisted of the 41 patients with diabetic foot ulcer older than 20 years, to whom the treatment with Heberprot-P. We analyzed: age, associated diseases, years of evolution of the disease, medication dose and healing. Results: female patients predominated (56.1%); the age group of 60-69 years (34.2%) and hypertension as an associated disease (48.8%); 43, 9% had an evolution of diabetes of 10-19 years; according to the Wagner classification, group III prevailed with 48.8% and II with 31.7%; 5 to 9 doses of Heberprot-P were applied in 43.5%. 78% achieved complete healing. Conclusions: in three quarters of patients with diabetic foot ulcer complete healing was achieved with the application of Heberprot-P therapy.</p

    Determinación del nivel de preparación metodológica del profesor para desarrollar tareas docentes integradoras en Morfofisiología

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    Fundamento: la tarea docente constituye la célula fundamental del proceso docente educativo, por lo que su confección y utilización constituyen premisas para el buen desempeño escolar.Objetivo: determinar el estado actual de la preparación metodológica del profesor para desarrollar tareas docentes integradoras en la disciplina Morfofisiología.Métodos: se realizó una investigación descriptiva transversal, la población de estudio estuvo conformada por los profesores que imparten la disciplina Morfofisiología en el área sur de la Universidad de Ciencias Médicas de Ciego de Ávila. Se aplicaron métodos teóricos: histórico-lógico, analítico-sintético e inductivo-deductivo; empíricos: análisis documental, observación a clases y encuesta en forma de cuestionario a docentes y de entrevista a informantes clave.Resultados: la tarea docente integradora no ha constituido un elemento de prioridad en la preparación del profesor, algunos no tienen en cuenta el algoritmo a seguir durante sus etapas de diseño, orientación, ejecución y control; existe insuficiente continuidad de temas afines y algunas insuficiencias en el dominio de contenidos, sobre todo de aquellos que no se relacionan directamente con las especialidades de los docentes.Conclusiones: las limitaciones identificadas para el desarrollo de tareas docentes afectan la integración que deben tener las asignaturas dentro de la propia disciplina Morfofisiología y con la disciplina rectora Medicina General Integral, para lograr un enfoque interdisciplinario en la formación del estudiante de las ciencias médicas. 

    In Vitro Effect of &Delta;9-Tetrahydrocannabinol and Cannabidiol on Cancer-Associated Fibroblasts Isolated from Lung Cancer

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    There is evidence that demonstrates the effect of cannabinoid agonists inhibiting relevant aspects in lung cancer, such as proliferation or epithelial-to-mesenchymal transition (EMT). Most of these studies are based on evidence observed in in vitro models developed on cancer cell lines. These studies do not consider the complexity of the tumor microenvironment (TME). One of the main components of the TME is cancer-associated fibroblasts (CAFs), cells that are relevant in the control of proliferation and metastasis in lung cancer. In this work, we evaluated the direct effects of two cannabinoid agonists, tetrahydrocannabinol (THC) and cannabidiol (CBD), used alone or in combination, on CAFs and non-tumor normal fibroblasts (NFs) isolated from adenocarcinoma or from healthy lung tissue from the same patients. We observed that these compounds decrease cell density in vitro and inhibit the increase in the relative expression of type 1 collagen (COL1A1) and fibroblast-specific protein 1 (FSP1) induced by transforming growth factor beta (TGF&beta;). On the other hand, we studied whether THC and CBD could modulate the interactions between CAFs or NFs and cancer cells. We conditioned the culture medium with stromal cells treated or not with THC and/or CBD and cultured A549 cells with them. We found that culture media conditioned with CAFs or NFs increased cell density, induced morphological changes consistent with EMT, inhibited cadherin-1 (CDH1) gene expression, and induced an increase in the relative expression of cadherin-2 (CDH2) and vimentin (VIM) genes in A549 cells. These changes were inhibited or decreased by THC and CBD administered alone or in combination. In another series of experiments, we conditioned culture media with A549 cells treated or not with THC and/or CBD, in the presence or absence of TGF&beta;. We observed that culture media conditioned with A549 in the presence of TGF&beta; induced an increase in the expression of COL1A1 and VIM, both in CAFs and in non-tumor NFs. Both THC and CBD ameliorated these effects. In summary, the results presented here reinforce the usefulness of cannabinoid agonists for the treatment of some relevant aspects of lung cancer pathology, and demonstrate in a novel way their possible effects on CAFs as a result of their relationship with cancer cells. Likewise, the results reinforce the usefulness of the combined use of THC and CBD, which has important advantages in relation to the possibility of using lower doses, thus minimizing the psychoactive effects of THC

    New roles for AP-1/JUNB in cell cycle control and tumorigenic cell invasion via regulation of cyclin E1 and TGF-β2

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    International audienceAbstract Background JUNB transcription factor contributes to the formation of the ubiquitous transcriptional complex AP-1 involved in the control of many physiological and disease-associated functions. The roles of JUNB in the control of cell division and tumorigenic processes are acknowledged but still unclear. Results Here, we report the results of combined transcriptomic, genomic, and functional studies showing that JUNB promotes cell cycle progression via induction of cyclin E1 and repression of transforming growth factor (TGF)-β2 genes. We also show that high levels of JUNB switch the response of TGF-β2 stimulation from an antiproliferative to a pro-invasive one, induce endogenous TGF-β2 production by promoting TGF-β2 mRNA translation, and enhance tumor growth and metastasis in mice. Moreover, tumor genomic data indicate that JUNB amplification associates with poor prognosis in breast and ovarian cancer patients. Conclusions Our results reveal novel functions for JUNB in cell proliferation and tumor aggressiveness through regulation of cyclin E1 and TGF-β2 expression, which might be exploited for cancer prognosis and therapy
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