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The administration of extracellular vesicles (EV) from mesenchymal stromal cells (MSC) is a promising cell-free nanotherapy for tissue repair after myocardial infarction (MI). However, the optimal EV delivery strategy remains undetermined. Here, we designed a novel MSC-EV delivery, using 3D scaffolds engineered from decellularised cardiac tissue as a cell-free product for cardiac repair. EV from porcine cardiac adipose tissue-derived MSC (cATMSC) were purified by size exclusion chromatography (SEC), functionally analysed and loaded to scaffolds. cATMSC-EV markedly reduced polyclonal proliferation and pro-inflammatory cytokines production (IFNγ, TNFα, IL12p40) of allogeneic PBMC. Moreover, cATMSC-EV recruited outgrowth endothelial cells (OEC) and allogeneic MSC, and promoted angiogenesis. Fluorescently labelled cATMSC-EV were mixed with peptide hydrogel, and were successfully retained in decellularised scaffolds. Then, cATMSC-EV-embedded pericardial scaffolds were administered in vivo over the ischemic myocardium in a pig model of MI. Six days from implantation, the engineered scaffold efficiently integrated into the post-infarcted myocardium. cATMSC-EV were detected within the construct and MI core, and promoted an increase in vascular density and reduction in macrophage and T cell infiltration within the damaged myocardium. The confined administration of multifunctional MSC-EV within an engineered pericardial scaffold ensures local EV dosage and release, and generates a vascularised bioactive niche for cell recruitment, engraftment and modulation of short-term post-ischemic inflammation., Graphical abstract Image 1, Highlights • A bioactive engineered scaffold can locally deliver EV to the infarcted myocardium. • Porcine cATMSC-EV promote endothelial cell recruitment and in vivo vascularization. • Porcine cATMSC-EV reduce inflammation, and macrophage and T cell infiltration.

Background Coronary artery disease (CAD) is the leading cause of death worldwide, and percutaneous coronary intervention with stenting the most widely performed procedure to treat CAD. However, current stent monitoring techniques are invasive and/or ionizing. Microwave spectrometry (MWS) may provide a non-invasive, non-ionizing and cost-effective alternative capable of detecting stent-related pathologies before fatal heart failure. Purpose To develop a new MWS-based technology to detect coronary stents in an in vivo swine model. Methodology First, using two new MWS devices, an in vitro experiment was carried out to demonstrate their ability of detecting the presence of: (1) a stent and (2) stent fractures (SF). To that end, an intact stent was distanced 3, 7, 11 and 15 mm from a MWS near-field probe in open-air conditions. Afterwards, three identical stents were piecemeal cut to emulate type I, II and III SF at different fractions of the stent's length (l): l/5, l/3 or l/2. Additionally, the stent was measured in a phantom substance, to simulate in vivo conditions: it was distanced from 0 to 40 mm in steps of 5 mm. Likewise, using a pair of MWS far-field antennas, the stent in phantom was measured at 10, 20, 30 and 40 mm. Finally, the MWS technology was assessed in vivo. To that end, six Landrace X Large White pigs were submitted to a stent implantation into the circumflex coronary artery (CX). The antennas measured the stent non-invasively, over the rib cage of the animals. MWS analysis were performed baseline (before stent implantation), and at 0, 3, 7, 14, 21 and 35 days of follow up. Measurements were performed only before ventricular systole to avoid differences in the stent position and deformation. Results In vitro, maxima and minima extrema in the microwave frequency response (see figure) were used to detect the stent. Type I and II SF produced 5 and 10% downshifts in the extrema frequencies with respect to the baseline values (unbroken stent), while type III produced 20% upshifts and a maxima splitting. Embedding the stent in phantom produced 25% downshift in the extrema frequencies. In vivo, the MWS antennas were useful to detect the stent presence into the CX artery during all time points of study, in all animals. Conclusions We have developed a new non-invasive and non-ionizing MWS technology capable of detecting the presence of a stent in a porcine model. Furthermore, we have proven how our technology can monitor structural damages in the stent (SF) and changes in its environment. This study proves the MWS potential to become a simple and yet effective method to arise stent-related pathologies in a pre-clinical stage; it could also provide physical insight about additional biological processes. Further improvements on the MWS device as well as in vivo measurements will ensure its consistency when monitoring human stents. Stent detection using MWS technique Funding Acknowledgement Type of funding source: Foundation. Main funding source(s): Fundaciό La MARATÓ de TV3, Generalitat de Catalunya, Red de Terapia Celular – TerCel, CIBER Cardiovascular, Spanish Agencia Estatal de Investigaciόn Unidad de Excelencia Maria de Maeztu, Sociedad Española de Cardilogía

Dentro de las enzimas antimicrobianas reportadas, las lisozimas y las enzimas tipo lisozimas procedentes de diversas fuentes naturales han mostrado actividad antibacteriana y antifúngica, haciéndolas atractivas como nuevas alternativas para controlar los microorganismos patógenos. Aunque las enzimas antimicrobianas se han propuesto ampliamente en la industria alimentaria y en la protección de cultivos, su uso implica algunas desventajas, como la inactivación por inhibidores o productos químicos o condiciones de procesamiento, los altos costos de producción y purificación, y problemas de solubilidad e inestabilidad. La tecnología de inmovilización enzimática es una herramienta prometedora capaz de reducir estos inconvenientes, particularmente cuando está destinada a trabajar en medios sólidos. Esta revisión muestra el estado del arte sobre la actividad de la lisozima contra bacterias y hongos, centrándose en los mecanismos de acción involucrados. Además, también se discuten los aspectos y características más mportantes de las lisozimas y su posible aplicación tecnológica, incluida la aplicación de matrices poliméricas a base de quitosano para la inmovilización de las enzimas antimicrobianas.

El objetivo general del proyecto es aprovechar la madera de cafe para el desarrollo de un envase inocuo para fermentar bebidas alcoholicas. Los antecedentes consultados, sugieren que los analisis realizados a escala nanotecnologica para este tipo de material, no se han efectuado anteriormente. Por medio de la tecnica SEM para las muestras organicas no conductoras, se deben limpiar cuidadosamente, para que haya desprendimiento de electrones y sean captados por los sensores especificos del microscopio. Esta tecnica es capaz de generar imagenes de alta resolucion, lo que permite examinar caracteristicas espacialmente cercanas en la muestra, con una gran magnificacion y grado de detalle, debido a que la superficie del material es barrida con electrones, que son acelerados a traves de un canon. El microscopio, dispone de diversos sistemas de deteccion, con los que es posible diferenciar la senal producida por los electrones primarios (PE) y secundarios (SE). Los electrones secundarios son aquellos que se emplean generalmente para obtener una imagen realista de la superficie que estamos analizando y determinar su morfologia. Con este tipo de analisis en la madera de cafe, se espera obtener caracteristicas especificas de la composicion elemental, asi como las fisico-morfologicas, que le aportarian propiedades diferenciales a los productos derivados. La importancia de este estudio, radica en aprovechar mas de 1300 toneladas de residuos organicos, para los que alternativas convencionales como compostaje y venta para cocinar con lena, son insuficientes para el manejo efectivo de los volumenes de desechos de los procesos productivos. En algunos casos, no se encuentran opciones viables para el manejo de la madera de cafe, ademas de alejar a la empresa de su actividad economica principal, no generar productos con valor agregado y utilidades significativas, desconociendo el valor industrial y comercial que aun poseen, una vez se les ha dado una disposicion final en grandes cantidades de basura organica.

RESUMEN: Esta es una reflexión del carácter pedagógico y didáctico de las acciones educativas de los programas recreo-deportivos que se ofrecen hoy en el país y que nace de la preocupación por hacer evidente aquello que hace del deporte y la recreación un escenario formativo en cada una de sus prácticas. Esta problemática se ve materializada en la presencia de programas con enfoques tecnicistas que pueden llevar a un olvido de la experiencia y la sensibilidad, anulando otras formas de relaciones y generando una necesidad exclusiva de producción, rendimiento, organicidad y cosificación del cuerpo, en una sociedad que hoy más que nunca reclama prácticas corporales con un enfoque transformativo que aporte a la construcción de una sociedad en paz. Pretendemos llamar la atención sobre las concepciones didácticas y pedagógicas que acompaña hoy dichos escenarios y motivar así al surgimiento de nuevas propuestas necesarias para seguir construyendo la educación física. ABSTRACT: This is a reflection of pedagogical and didactic nature of the educational activities of recreation-sports programs offered today in the country and that stems from the concern to make clear what makes the sport and recreation a formative stage in each of their practices. This problem is embodied in the presence of specialized programs approaches that can lead to forgetting the experience and sensitivity, negating other forms of relationships and generating an exclusive production needs, efficiency, organic and objectification of the body, in a society now claims bodily practices with a transformative approach that contributes to building a peaceful society. We intend to draw attention to the didactics and pedagogical concepts that accompanies such scenarios today and so encourage the emergence of new proposals necessary to continue building Physical Education. COL0043093

Farmed shrimp Litopenaeus vannamei, one of the most consumed seafood in the planet, is a source of antiproliferative extracts still to be fully characterized. This study the isolation and identification of these antiproliferative compounds. From a chloroform-soluble extract from shrimp muscle, hexane- and methanol-soluble fractions were obtained and tested for antiproliferative activity (MTT), a bioassay that guided the fractionation and isolation of bioactive fractions using open column chromatography. MeOH-soluble fraction resulted bioactive and was subjected to further fractionation from which one subfraction outstand for being highly active against prostate cancer cell line. Antiproliferative effects were evaluated using colorimetric assays and cell morphology observations. Further chromatographic procedures resulted in sub-fractions from which one was effective in causing DNA damage and F-actin polymerization, which suggests cellular collapse and apoptosis. According to the structural chemical characterization carried out, dioctyl phthalate, eicosapentaenoic acid, and an indolocarbazole alkaloid type of compound were identified. This last compound, which resulted majorly responsible for the bioactivity, was not found reported in the available databases. Pure EPA control was used to compare it with the subfraction, observing greater activity in the subfraction than when EPA was used, suggesting that another compound different from EPA is providing the highest activity; but, more investigation is needed for a full chemical and structural characterization.

Type I collagen hydrogels are of high interest in tissue engineering. With the evolution of 3D bioprinting technologies, a high number of collagen-based scaffolds have been reported for the development of 3D cell cultures. A recent proposal was to mix collagen with silk fibroin derived from Bombyx mori silkworm. Nevertheless, due to the difficulties in the preparation and the characteristics of the protein, several problems such as phase separation and collagen denaturation appear during the procedure. Therefore, the common solution is to diminish the concentration of collagen although in that way the most biologically relevant component is reduced. In this study, we present a new, simple, and effective method to develop a collagen-silk hybrid hydrogel with high collagen concentration and with increased stiffness approaching that of natural tissues, which could be of high interest for the development of cardiac patches for myocardial regeneration and for preconditioning of mesenchymal stem cells (MSCs) to improve their therapeutic potential. Sericin in the silk was preserved by using a physical solubilizing procedure that results in a preserved fibrous structure of type I collagen, as shown by ultrastructural imaging. The macro- and micromechanical properties of the hybrid hydrogels measured by tensile stretch and atomic force microscopy, respectively, showed a more than twofold stiffening than the collagen-only hydrogels. Rheological measurements showed improved printability properties for the developed biomaterial. The suitability of the hydrogels for 3D cell culture was assessed by 3D bioprinting bone marrow-derived MSCs cultured within the scaffolds. The result was a biomaterial with improved printability characteristics that better resembled the mechanical properties of natural soft tissues while preserving biocompatibility owing to the high concentration of collagen. Impact statement In this study, we report the development of silk microfiber-reinforced type I collagen hydrogels for 3D bioprinting and cell culture. In contrast with previously reported studies, a novel physical method allowed the preservation of the silk sericin protein. Hydrogels were stable, showed no phase separation between the biomaterials, and they presented improved printability. An increase between two- and threefold of the multiscale stiffness of the scaffolds was achieved with no need of using additional crosslinkers or complex methods, which could be of high relevance for cardiac patches development and for preconditioning mesenchymal stem cells (MSCs) for therapeutic applications. We demonstrate that bone marrow-derived MSCs can be effectively bioprinted and 3D cultured within the stiffened structures. This work was supported in part by the Spanish Ministry of Sciences, Innovation and Universities (DPI2017-83721-P and PGC2018-097323-A-I00) and by the Marie Sklodowska- Curie Action, Innovative Training Networks 2018, EU Grant Agreement no. 812772.

Insulin-like growth factor-1 (IGF-1) has demonstrated beneficial effects after myocardial infarction (MI). Microencapsulation of IGF-1 could potentially improve results. We aimed to test the effect of an intracoronary (IC) infusion of microencapsulated IGF-1 in a swine acute MI model. For that purpose IC injection of a 10 ml solution of 5 × 106 IGF-1 loaded microspheres (MSPs) (n = 8, IGF-1 MSPs), 5 × 106 unloaded MSPs (n = 9; MSPs) or saline (n = 7; CON) was performed 48 hours post-MI. Left ventricular ejection fraction (LVEF), indexed ventricular volumes and infarct size (IS) were determined by cardiac magnetic resonance at pre-injection and 10 weeks. Animals were euthanized at 10 weeks, and myocardial fibrosis and vascular density were analysed. End-study LVEF was significantly greater in IGF-1 MSPs compared to MSPs and CON, while ventricular volumes exhibited no significant differences between groups. IS decreased over time in all groups. Collagen volume fraction on the infarct area was significantly reduced in IGF-1 MSPs compared to CON and MSPs. Vascular density analysis of infarct and border zones showed no significant differences between groups. In conclusion, the IC injection of 5 × 106 IGF-1 loaded MSPs in a porcine acute MI model successfully improves cardiac function and limits myocardial fibrosis, which could be clinically relevant.

The oxidative stress damage on cells is an example of roles in the pathogenesis of different degenerative diseases and the search of compounds that can slow this oxidation is continuous. The aim of this work was to obtain bioactive fractions from wild shrimp (Litopenaeus stylirostris) muscle, in order to evaluate their protective capacity and chemo-structurally characterize them. ABTS and DPPH, and FRAP assays suggested that bioactive fractions possess free radical-scavenging capacity, and reducing power, respectively. An inhibitory effect observed on AAPH-induced hemolysis and results from an H2O2-derived radicals-scavenging assay (retinoprotective) suggest that these fractions can exert their protective activity in human cells. UV–Vis, fluorescence, 13C-, 1H-NMR, and ESI-MS studies performed on the most bioactive fraction, suggests that their main components are eicosapentaenoic acid, dioctyl phthalate, and a possibly novel indolocarbazole alkaloid derivative. These results suggest that these compounds are good candidates to further investigations as possible chemoprotective agents.

Summary Myocardial infarction (MI) remains a dreadful disease around the world, causing irreversible sequelae that shorten life expectancy and reduce quality of life despite current treatment. Here, the authors engineered a cell-enriched myocardial graft, composed of a decellularized myocardial matrix refilled with adipose tissue-derived progenitor cells (EMG-ATDPC). Once applied over the infarcted area in the swine MI model, the EMG-ATDPC improved cardiac function, reduced infarct size, attenuated fibrosis progression, and promoted neovascularization of the ischemic myocardium. The beneficial effects exerted by the EMG-ATDPC and the absence of identified adverse side effects should facilitate its clinical translation as a novel MI therapy in humans., Visual Abstract, Highlights • MI remains a major cause of morbidity and mortality despite major treatment advances achieved during the past decades. • Administration of an engineered myocardial graft, composed of decellularized myocardial matrix refilled with ATDPCs (EMG-ATDPC), in a porcine pre-clinical MI model, may support cardiac recovery following MI. • Thirty days post-EMG-ATDPC implantation, cardiac magnetic resonance imaging and comprehensive histological analysis were performed to evaluate its impact on myocardial restoration. • EMG-ATDPC resulted in better left ventricular ejection fraction, higher vessel density and neovascularization, and reduced infarct size by 68%, as well as limited fibrosis. • Accordingly, EMG-ATDPC is ready to start the translational avenue toward phase I first-in-man clinical trials.

There is increasing interest in identifying novel stem cell sources for application in emerging cell therapies. In this context, umbilical cord blood (UCB) shows great promise in multiple clinical settings. The number of UCB banks has therefore increased worldwide, with the objective of preserving potentially life-saving cells that are usually discarded after birth. After a rather long and costly processing procedure, the resultant UCB-derived cell products are cryopreserved until transplantation to patients. However, in many cases, only a small proportion of administered cells engraft successfully. Thus, can we do any better regarding current UCB-based therapeutic approaches? Here we discuss concerns about the use of UCB that are not critically pondered by researchers, clinicians, and banking services, including wasting samples with small volumes and the need for more reliable quality and functional controls to ensure the biological activity of stem cells and subsequent engraftment and treatment efficacy. Finally, we appeal for collaborative agreements between research institutions and UCB banks in order to redirect currently discarded small-volume UCB units for basic and clinical research purposes. Developmental Dynamics 245:710-717, 2016. © 2016 Wiley Periodicals, Inc.

Fibrin is a topical haemostat, sealant and tissue glue, which consists of concentrated fibrinogen and thrombin. It has broad medical and research uses. Recently, several studies have shown that engineered patches comprising mixtures of biological or synthetic materials and progenitor cells showed therapeutic promise for regenerating damaged tissues. In that context, fibrin maintains cell adherence at the site of injury, where cells are required for tissue repair, and offers a nurturing environment that protects implanted cells without interfering with their expected benefit. Here we review the past, present and future uses of fibrin, with a focus on its use as a scaffold material for cardiac repair. Fibrin patches filled with regenerative cells can be placed over the scarring myocardium; this methodology avoids many of the drawbacks of conventional cell-infusion systems. Advantages of using fibrin also include extraction from the patient's blood, an easy readjustment and implantation procedure, increase in viability and early proliferation of delivered cells, and benefits even with the patch alone. In line with this, we discuss the numerous preclinical studies that have used fibrin-cell patches, the practical issues inherent in their generation, and the necessary process of scaling-up from animal models to patients. In the light of the data presented, fibrin stands out as a valuable biomaterial for delivering cells to damaged tissue and for promoting beneficial effects. However, before the fibrin scaffold can be translated from bench to bedside, many issues must be explored further, including suboptimal survival and limited migration of the implanted cells to underlying ischaemic myocardium. Copyright © 2016 John Wiley & Sons, Ltd.

Demuestra el incremento en la tasa de retención de clientes a partir del desarrollo de estrategias de marketing adecuadas en el servicio postventa y la utilización de técnicas de inbound marketing para atraer y aumentar el tráfico de clientes. La investigación hace uso de un método mixto con la realización de 21 encuestas y 1 entrevista a especialistas de marketing, servicio postventa y CRM en las tiendas por departamento que operan en el Perú. La investigación demuestra que la aplicación combinada de las dos mejoras propuestas generan un incremento en la tasa de retención de clientes en orden de 58.8%, pasando así dicha tasa de un 51% a 80.9%.

Altres ajuts: Fundació La MARATÓ de TV3 (20153530/31), Unidad de Excelencia Maria de Maeztu (MDM2016-0600). This work has been developed in the context of AdvanceCat with the support of ACCIÓ (Catalonia Trade & Investment; Generalitat de Catalunya) Some conditions are well known to be directly associated with stent failure, including in-stent re-occlusion and stent fracture. Currently, identification of these high-risk conditions requires invasive and complex procedures. This study aims to assess microwave spectrometry (MWS) for monitoring stents non-invasively. Preliminary ex vivo data are presented to move to in vivo validation. Fifteen mice were assigned to receive subcutaneous stent implantations (n = 10) or sham operations (n = 5). MWS measurements were carried out at 0, 2, 4, 7, 14, 22, and 29 days of follow-up. Additionally, 5 stented animals were summited to micro-CT analyses at the same time points. At 29 days, 3 animals were included into a stent fracture subgroup and underwent a last MWS and micro-CT analysis. MWS was able to identify stent position and in-stent stenosis over time, also discerning significant differences from baseline measures (P < 0.001). Moreover, MWS identified fractured vs. non-fractured stents in vivo. Taken together, MWS emerges as a non-invasive, non-ionizing alternative for stent monitoring. MWS analysis clearly distinguished between in-stent stenosis and stent fracture phenomena.

A wide variety of chitosan (CS) biomaterials have been loaded with different antimicrobial agents to improve the activity of CS against phytopathogenic fungi. Recently, the antimicrobial activity of 1H-pyrrole-2-carboxylic acid (PCA) has been reported as a secondary metabolite of Streptomyces griseus, which was identified as the main bioactive compound in the biological control. However, it is sensitive to light and its activity against filamentous fungi has not yet been reported. The aim of the present research work was to evaluate the biological activity of CS-PCA biocomposites for the control of Aspergillus niger. CS-PCA biocomposites were obtained through nanoprecipitation. In vitro antifungal activity was determined by viability assay, spore germination, morphometric analysis of spores and hyphae, and the analysis of cellular components by fluorescence microscopy. CS-PCA showed an average size and Z potential of 502 ± 72 nm and + 54.7 ± 15 mV, respectively. Micrographs demonstrated well-distributed biocomposites with an apparently spherical shape. A new signal at 1473 cm−1 in the FT-IR spectrum of the CS-PCA biocomposite was observed, confirming the presence of PCA in the composition of the CS-PCA nanosystem. CS-PCA biocomposites reduced the spores’ viability by up to 58%. Effects on fungi morphometry, observed as an increase in the spores’ average diameter, swelling, distortion, and an increase in the branching of hyphae, were observed. Fluorescence analysis showed oxidative stress and membrane and cell wall damage, mainly at early growth stages. The inhibitory effect against CS-resistant fungi, such as A. niger, opens a door for the control of CS-sensitive fungi.

Mechanical conditioning is incompletely characterized for stimulating therapeutic cells within the physiological range. We sought to unravel the mechanism of action underlying mechanical conditioning of adipose tissue-derived progenitor cells (ATDPCs), both in vitro and in silico. Cardiac ATDPCs, grown on 3 different patterned surfaces, were mechanically stretched for 7 days at 1 Hz. A custom-designed, magnet-based, mechanical stimulator device was developed to apply ~10% mechanical stretching to monolayer cell cultures. Gene and protein analyses were performed for each cell type and condition. Cell supernatants were also collected to analyze secreted proteins and construct an artificial neural network. Gene and protein modulations were different for each surface pattern. After mechanostimulation, cardiac ATDPCs increased the expression of structural genes and there was a rising trend on cardiac transcription factors. Finally, secretome analyses revealed upregulation of proteins associated with both myocardial infarction and cardiac regeneration, such as regulators of the immune response, angiogenesis or cell adhesion. To conclude, mechanical conditioning of cardiac ATDPCs enhanced the expression of early and late cardiac genes in vitro. Additionally, in silico analyses of secreted proteins showed that mechanical stimulation of cardiac ATDPCs was highly associated with myocardial infarction and repair.

Chronic diseases, including myocardial scar healing and heart failure remission, impose huge social and economic burdens, and novel approaches are needed. Several therapeutic modalities are currently being evaluated, including cell therapy, stem cell conditioning, and cardiac tissue engineering. Areas covered: This review discusses the restoration of cardiac function after myocardial infarction using a vascularized flap of autologous cardiac adipose tissue over an akinetic scar. It addresses the risks and benefits of using cardiac adipose progenitors and the adipose graft transposition procedure (AGTP) to ameliorate cardiac dysfunction in preclinical and clinical trials. Expert commentary: The focus is shifting from first-generation studies that used ex vivo expanded and manipulated progenitors to newer second-generation approaches, including AGTP, which are inexpensive and do not raise ethical issues. AGTP safety has been validated, and the ongoing AGTP-2 trial to determine AGTP efficacy and outcome is currently recruiting patients (NCT02798276). This reparative strategy is safe, avoids the risks associated with ex vivo manipulation, and the preclinical and clinical trials performed to date show cardiac function recovery and reduced necrosis. Confirmation of these data in the AGTP-2 trial could pave the way for the clinical use of this novel modality.

Dilated cardiomyopathy (DCM) remains the most frequent cause of cardiac transplant and thus results in an enormous cost burden for health care systems worldwide. Although DCM is thought to be induced mainly by genetic and/or environmental factors, the cause is unknown in the majority of cases, giving rise to the term idiopathic DCM. Marked cardiac endothelial changes are associated with disease progression and outcome, and there are ongoing efforts to identify biomarkers that have diagnostic and prognostic value. Here, we discuss the potential and the limitations of circulating endothelial progenitor cells (EPCs) as minimally invasive serological biomarkers for DCM. In this context, it is essential to further evaluate their clinical utility independently of other variable factors that can also affect EPC levels such as age, gender, lifestyles, and treatments. To that end, large multicenter studies and standardized instrument settings, reagents, and sample preparation protocols are needed to confirm this.

INTRODUCTION AND OBJECTIVES: Neprilysin breaks down numerous vasoactive peptides. The soluble form of neprilysin, which was recently identified in heart failure, is associated with cardiovascular outcomes. Within a multibiomarker strategy, we directly compared soluble neprilysin and N-terminal pro-B-type natriuretic peptide as risk stratifiers in a real-life cohort of heart failure patients. METHODS: Soluble neprilysin, N-terminal pro-B-type natriuretic peptide, ST2, and high-sensitivity troponin T levels were measured in 797 consecutive ambulatory heart failure patients followed up for 4.7 years. Comprehensive multivariable analyses and soluble neprilysin vs N-terminal pro-B-type natriuretic peptide head-to-head assessments of performance were performed. A primary composite endpoint included cardiovascular death or heart failure hospitalization. A secondary endpoint explored cardiovascular death alone. RESULTS: Median soluble neprilysin and N-terminal pro-B-type natriuretic peptide concentrations were 0.64ng/mL and 1187 ng/L, respectively. Both biomarkers significantly correlated with age (P

Resumen Introduccion y objetivos La neprilisina degrada numerosos peptidos vasoactivos. La forma soluble de neprilisina, que se ha identificado recientemente en la insuficiencia cardiaca, se asocia con eventos cardiovasculares. Se compararon de manera directa la neprilisina soluble y la fraccion aminoterminal del propeptido natriuretico cerebral como estratificadores de riesgo, dentro de una estrategia multimarcadores, en una cohorte de pacientes con insuficiencia cardiaca de la practica clinica real. Metodos Se determinaron las concentraciones de neprilisina soluble, la fraccion aminoterminal del propeptido natriuretico cerebral, ST2 y troponina T de alta sensibilidad en 797 pacientes ambulatorios consecutivos con insuficiencia cardiaca seguidos durante 4,7 anos. Se llevaron a cabo analisis multivariables exhaustivos y se realizaron comparaciones directas de neprilisina soluble frente a la fraccion aminoterminal del propeptido natriuretico cerebral mediante estadisticas de rendimiento. El objetivo final principal fue el compuesto por muerte cardiovascular u hospitalizacion por insuficiencia cardiaca. Un objetivo secundario exploro la muerte cardiovascular sola. Resultados Las medianas de concentracion de neprilisina soluble y la fraccion aminoterminal del propeptido natriuretico cerebral fueron de 0,64 ng/ml y 1.187 ng/l respectivamente. Ambos biomarcadores presentaron una correlacion significativa con la edad (p hazard ratio = 1,14; intervalo de confianza del 95%, 1,02-1,27; p = 0,03) y la muerte cardiovascular ( hazard ratio = 1,15; intervalo de confianza del 95%, 1,01-1,31; p = 0,04), pero no asi la fraccion aminoterminal del propeptido natriuretico cerebral. La comparacion directa de la neprilisina soluble con la fraccion aminoterminal del propeptido natriuretico cerebral puso de manifiesto buena calibracion y discriminacion y reclasificacion similares con ambos biomarcadores neurohormonales, pero solo la neprilisina soluble mejoro la bondad de ajuste global. Conclusiones La neprilisina soluble mantuvo el valor pronostico independiente al incluirlo en una estrategia multimarcadores, mientras que la fraccion aminoterminal del propeptido natriuretico cerebral perdio la significacion en la estratificacion del riesgo en los pacientes ambulatorios con insuficiencia cardiaca. Ambos biomarcadores obtuvieron medidas de rendimiento similares en los analisis de comparacion directa.

Considerable research has been dedicated to restoring myocardial cell slippage and limiting ventricular remodeling after myocardial infarction (MI). We examined the ability of a three-dimensional (3D) engineered fibrin patch filled with human umbilical cord blood-derived mesenchymal stem cells (UCBMSCs) to induce recovery of cardiac function after MI. The UCBMSCs were modified to coexpress luciferase and fluorescent protein reporters, mixed with fibrin, and applied as an adhesive, viable construct (fibrin-cell patch) over the infarcted myocardium in mice (MI-UCBMSC group). The patch adhered well to the heart. Noninvasive bioluminescence imaging demonstrated early proliferation and differentiation of UCBMSCs within the construct in the postinfarct mice in the MI-UCBMSC group. The implanted cells also participated in the formation of new, functional microvasculature that connected the fibrin-cell patch to both the subjacent myocardial tissue and the host circulatory system. As revealed by echocardiography, the left ventricular ejection fraction and fractional shortening at sacrifice were improved in MI-UCBMSC mice and were markedly reduced in mice treated with fibrin alone and untreated postinfarction controls. In conclusion, a 3D engineered fibrin patch composed of UCBMSCs attenuated infarct-derived cardiac dysfunction when transplanted locally over a myocardial wound. Significance Ischemic heart failure (HF) is the end stage of many cardiovascular diseases, including myocardial infarction. The only definitive treatment for HF is cardiac transplant, which is hampered by limited number of heart donors and graft rejection. In recent times, cellular cardiomyoplasty has been expected to repair infarcted myocardium by implantation of different sources of stem or progenitor cells. However, low cell survival and myocardial implantation rates have motivated the emergence of novel approaches with the objective of generating graftable cell-based implants. Here, the potential of 3D engineered fibrin-umbilical cord blood-derived mesenchymal stem cells patches is shown to significantly recover lost general functions in post-infarcted mice.

Cardiac tissue engineering is becoming a prime destination on the road map for translational medicine, often referred to as bench-to-bedside research [(1,2)][1]. It has been 20 years since the first cardiac tissue engineering papers were published; now more than 700 papers are added each year under

Altres ajuts: AB-G was supported by Grants from the Ministerio de Educación y Ciencia (SAF2014-59892), Fundació La MARATÓ de TV3 (201502, 201516), CIBER Cardiovascular (CB16/11/00403), and AdvanceCat with the support of ACCIÓ [Catalonia Trade & Investment; Generalitat de Catalunya] under the Catalonian ERDF [European Regional Development Fund] operational program, 2014-2020. Cross-sectional investigations report shorter telomeres in patients with heart failure (HF); however, no studies describe telomere length (TL) trajectory and its relationship with HF progression. Here we aimed to investigate telomere shortening over time and its relationship to outcomes. Our study cohort included 101 ambulatory patients with HF. Blood samples were collected at baseline (n = 101) and at the 1-year follow-up (n = 54). Using flow-FISH analysis of circulating monocytes, we simultaneously measured three monocyte subsets-classical (CD14 ++ CD16 −), intermediate (CD14 ++ CD16 +), and nonclassical (CD14 + CD16 ++)-and their respective TLs based on FITC-labeled PNA probe hybridization. The primary endpoints were all-cause death and the composite of all-cause death or HF-related hospitalization, assessed at 2.3 ± 0.6 years. All statistical analyses were executed by using the SPSS 15.0 software, and included Student's t test and ANOVA with post hoc Scheffe analysis, Pearson or Spearman rho correlation and univariate Cox regression when applicable. We found high correlations between TL values of different monocyte subsets: CD14 ++ CD16 + vs. CD14 ++ CD16 −, R = 0.95, p < 0.001; CD14 ++ CD16 + vs. CD14 + CD16 ++, R = 0.90, p < 0.001; and CD14 ++ CD16 − vs. CD14 + CD16 ++, R = 0.89, p < 0.001. Mean monocyte TL exhibited significant attrition from baseline to the 1-year follow-up (11.1 ± 3.3 vs. 8.3 ± 2.1, p < 0.001). TL did not significantly differ between monocyte subsets at either sampling time-point (all p values > 0.1). Cox regression analyses did not indicate that TL or ΔTL was associated with all-cause death or the composite endpoint. Overall, this longitudinal study demonstrated a ~ 22% reduction of TL in monocytes from ambulatory patients with HF within 1 year. TL and ΔTL were not related to outcomes over long-term follow-up. The online version of this article (10.1186/s12967-018-1412-z) contains supplementary material, which is available to authorized users.

Idiopathic dilated cardiomyopathy (IDCM) is a frequent cause of heart transplantation. Potentially valuable blood markers are being sought, andlow-density lipoprotein receptor-related protein 1 (LRP1) has been linked to the underlying molecular basis of the disease. This study comparedcirculating levels of soluble LRP1 (sLRP1) in IDCM patients and healthy controls and elucidated whether sLRP1 is exported out of the myocardiumthrough extracellular vesicles (EVs) to gain a better understanding of the pathogenesis of the disease. LRP1achain expression was analysed insamples collected from the left ventricles of explanted hearts using immunohistochemistry. sLRP1 concentrations were determined in platelet-freeplasma by enzyme-linked immunosorbent assay. Plasma-derived EVs were extracted by size-exclusion chromatography (SEC) and characterizedby nanoparticle tracking analysis and cryo-transmission electron microscopy. The distributions of vesicular (CD9, CD81) and myocardial (caveolin-3) proteins and LRP1achain were assessed in SEC fractions by flow cytometry. LRP1achain was preferably localized to blood vessels in IDCMcompared to control myocardium. Circulating sLRP1 was increased in IDCM patients. CD9- and CD81-positive fractions enriched with membranevesicles with the expected size and morphology were isolated from both groups. The LRP1achain was not present in these SEC fractions, whichwere also positive for caveolin-3. The increase in circulating sLRP1 in IDCM patients may be clinically valuable. Although EVs do not contribute tohigher sLRP1 levels in IDCM, a comprehensive analysis of EV content would provide further insights into the search for novel blood markers, This work was supported by grants from the Ministerio de Educación y Ciencia (SAF2014-59892-R), the Ministerio de Econom ıa yCompetitividad (Juan de la Cierva, JCI-2012-14025), Fundacio La MARATO de TV3 (201502, 201516, 201521_10), the Fundaci o Daniel Bravo Andreu, the Sociedad Española de Cardiología, the Societat Catalana de Cardiologia, the Generalitat de Catalunya (SGR 2014,CERCA Programme), and the Fundacio Bancaria La Caixa. This workwas also funded by the Red de Terapia Celular–TerCel (RD16/0011/0006), the CIBER Cardiovascular–(CB16/11/00403) and Fondo deInvestigaci on Sanitaria, Instituto de Salud Carlos III (FIS PI14/01682,FIS PI14/01729 and CD14/00109) as part of the Plan Nacional deI+D+I cofounded by ISCIII-Sudirecci on General de Evaluaci on y elFondo Europeo de Desarrollo Regional (FEDER)

For years, sufficient progress has been made in treating heart failure following myocardial infarction; however, the social and economic burdens and the costs to world health systems remain high. Moreover, treatment advances have not resolved the underlying problem of functional heart tissue loss. In this field of research, for years we have actively explored innovative biotherapies for cardiac repair. Here, we present a general, critical overview of our experience in using mesenchymal stem cells, derived from cardiac adipose tissue and umbilical cord blood, in a variety of cell therapy and tissue engineering approaches. We also include the latest advances and future challenges, including good manufacturing practice and regulatory issues. Finally, we evaluate whether recent approaches hold potential for reliable translation to clinical trials.

The combination of biomatrices and induced pluripotent stem cell (iPSC) derivatives to aid repair and myocardial scar formation may soon become a reality for cardiac regenerative medicine. However, the tumor risk associated with residual undifferentiated cells remains an important safety concern of iPSC-based therapies. This concern is not satisfactorily addressed in xenotransplantation, which requires immune suppression of the transplanted animal. In this study, we assessed the safety of transplanting undifferentiated iPSCs in an allogeneic setting. Given that swine are commonly used as large animal models in cardiac medicine, we used porcine iPSCs (p-iPSCs) in conjunction with bioengineered constructs that support recovery after acute myocardial infarction. Histopathology analyses found no evidence of p-iPSCs or p-iPSC-derived cells within the host myocardium or biomatrices after 30 and 90 days of follow-up. Consistent with the disappearance of the implanted cells, we could not observe functional benefit of these treatments in terms of left ventricular ejection fraction, cardiac output, ventricular volumes, or necrosis. We therefore conclude that residual undifferentiated iPSCs should pose no safety concern when used on immune-competent recipients in an allogeneic setting, at least in the context of cardiac regenerative medicine.

Dilated cardiomyopathy is the most frequent etiology of non-ischemic heart failure. In a majority of cases the causal mechanism is unknown, giving rise to the term ‘idiopathic' dilated cardiomyopathy (IDCM). Major pathological derangements include patchy interstitial fibrosis, degenerated cardiomyocytes, and dilatation of the cardiac chambers, but recent evidence suggests that disease progression may also have the signature of cardiac endothelial dysfunction. As we better understand the molecular basis of IDCM, novel therapeutic approaches, mainly gene transfer and cell-based therapies, are being explored. Cells with regenerative potential have been extensively tested in cardiac diseases of ischemic origin in both pre-clinical and clinical settings. However, whether cell therapy has any clinical value in IDCM patients is still being evaluated. This article is a concise summary of cell therapy studies for IDCM, with a focus on recent advances that highlight the vascular potential exhibited by umbilical cord blood-derived mesenchymal stem cells (UCBMSCs). We also provide an overview of cardiac vasculature as a key regulator of subjacent myocardial integrity and function, and discuss the potential mechanisms of UCBMSC amelioration of IDCM myocardium. Consideration of these issues shows that these cells are conceivably new therapeutic agents for this complex and elusive human disorder.

Objective— Hypoxia disturbs vascular function by promoting extracellular matrix remodeling. Extracellular matrix integrity and composition are modulated by metalloproteinases (MMPs). Our aim was to investigate the role of low-density lipoprotein receptor–related protein 1 (LRP1) in regulating MMP-9/MMP-2 activation and vascular smooth muscle cells (VSMCs) migration in response to hypoxia, and to elucidate the LRP1-signaling pathways involved in this process. Approach and Results— Western blot analysis showed that hypoxia induced a sustained phosphorylation of proline-rich tyrosine kinase 2 concomitantly with LRP1 overexpression in human VSMCs (hVSMCs). Deletion of LRP1 using small-interfering RNA technology or treatment of hVSMCs with the Src family kinase inhibitor PP2 impaired hypoxia-induced phosphorylation of proline-rich tyrosine kinase 2 levels. Coimmunoprecipitation experiments showed that the higher amounts of phosphorylation of proline-rich tyrosine kinase 2/LRP1β immunoprecipitates in hypoxic hVSMCs were abolished in PP2-treated hVSMCs. Both LRP1 silencing and PP2 treatment were highly effective in the prevention of hypoxia-induced MMP-9 activation and hVSMC migration. Cellular subfractionation experiments revealed that PP2 effects may be caused by impairment of hypoxia-induced nuclear factor-κβ translocation to the nucleus. ELISA measurements showed that LRP1 silencing but not PP2 treatment increased interleukin-1β, interleukin-6, and monocyte chemoattractant protein-1 secretion by hypoxic hVSMCs. Conclusions— Our findings determine a crucial role of LRP1-mediated Pyk2 phosphorylation on hypoxia-induced MMP-9 activation and hVSMC migration and therefore in hypoxia-induced vascular remodeling. Both LRP1 silencing and PP2 treatments also influence hypoxia-induced proinflammatory effects in hVSMCs. Therefore, further studies are required to establish therapeutical strategies that efficiently modulate vascular remodeling and inflammation associated with hypoxia-vascular diseases.

Objective: Idiopathic dilated cardiomyopathy (IDCM) affects myocardial vascularization. Whether a lack of demand for increased myocardial vascularization and/or an impaired response of circulating angiogenic-supportive cells are responsible for the vascular derangements found in IDCM is unknown. Methods and results: Left ventricle (LV) samples obtained at transplant from IDCM hearts were compared to control hearts from non-cardiac decedents. Peripheral colony-forming myeloid cells were extracted from age- and sex-matched IDCM patients and healthy volunteers. At the tissue level, no differences were detected in stromal cell-derived factor (SDF)-1 alpha expression, but integrin-linked kinase (ILK) levels and activity were increased in IDCM. A marked co-localization of SDF-1 alpha and the specific marker of cholesterol-enriched lipid rafts Flotillin (Flot)-1 was found in IDCM. SDF-1 alpha was also highly distributed into IDCM lipid rafts. Non-adherent pro-angiogenic cells from both groups, which were found increased in patients but showed similar surface levels of CXCR-4, equally supported Matrigel-mediated cell network formation. However, SDF-1-mediated migration was reduced in IDCM-derived cells, which also exhibited decreased ILK activity and downstream ERK activation. Conclusions: Taken together, our results point out that myocardial competency to increase vascularization is not altered in IDCM, but dysfunctional SDF-1-mediated migration by peripheral pro-angiogenic cells through ILK and downstream ERK signaling may compromise endothelial recovery in patients. We provide new insights into lipid raft function in human IDCM and envision more effective treatments. (C) 2013 Elsevier Ireland Ltd. All rights reserved.

The human heart has limited regenerative capacity. We focused on cardiac adipose tissue as a source of progenitor cells and biological matrix material for salvaging injured myocardium. First, a population of human adult mesenchymal-like progenitors derived from cardiac adipose tissue, with inherent cardiac and endothelial cell potential, was identified and characterized. Next, a salvage strategy was tested, where a pericardial-derived, vascularized, adipose flap was used to cover oxygen-deprived myocardium in a porcine model. The fat flap reduced the myocardial scar size, in both acute and chronic infarcts. A human clinical trial to examine this novel intervention is currently underway.

Heart failure is the end-stage of many cardiovascular diseases—such as acute myocardial infarction—and remains one of the most appealing challenges for regenerative medicine because of its high incidence and prevalence. Over the last 20 years, cardiomyoplasty , based on the isolated administration of cells with regenerative capacity, has been the focal point of most studies aimed at regenerating the heart. Although this therapy has proved feasible in the clinical setting, the degree of infarcted myocardium regenerated and of improved cardiac function are at best modest. Hence, tissue engineering has emerged as a novel technology using cells with regenerative capacity, biological and/or synthetic materials, growth, proangiogenic and differentiation factors, and online registry systems, to induce the regeneration of whole organs or locally damaged tissue. The next step, seen recently in pioneering animal studies, is de novo generation of bioartificial hearts by decellularization and preservation of supporting structures for their subsequent repopulation with new contractile, vascular muscle tissue. Ultimately, this new approach would entail transplantation of the “rebuilt” heart, reestablishing cardiac function in the recipient.

IntroductionCardiac adipose tissue is a source of progenitor cells with regenerative capacity. Studies in rodents demonstrated that the intramyocardial delivery of cells derived from this tissue improves cardiac function after myocardial infarction (MI). We developed a new reparative approach for damaged myocardium that integrates the regenerative properties of cardiac adipose tissue with tissue engineering. In the adipose graft transposition procedure (AGTP), we dissect a vascularised flap of autologous pericardial adipose tissue and position it over the myocardial scarred area. Following encouraging results in acute and chronic MI porcine models, we performed the clinical trial (NCT01473433, AdiFLAP trial) to evaluate safety in patients with chronic MI undergoing coronary artery bypass graft. The good safety profile and trends in efficacy warranted a larger trial.Study designThe AGTP II trial (NCT02798276) is an investigator initiated, prospective, randomised, controlled, multicentre study to assess the efficacy of the AGTP in 108 patients with non-revascularisable MI. Patients will be assigned to standard clinical practice or the AGTP. The primary endpoint is change in necrotic mass ratio by gadolinium enhancement at 91 and 365 days. Secondary endpoints include improvement in regional contractibility by MRI at 91 and 365 days; changes in functional MRI parameters (left ventricular ejection fraction, left and right ventricular geometric remodelling) at 91 and 365 days; levels of N-terminal prohormone of brain natriuretic peptide (NT-proBNP) at 7, 91 and 365 days; appearance of arrhythmias from 24 hour Holter monitoring at 24 hours, and at 91 and 365 days; all cause death or re-hospitalisation at 365 days; and cardiovascular death or re-hospitalisation at 365 days.Ethics and disseminationThe institutional review board approved the trial which will comply with the Declaration of Helsinki. All patients will provide informed consent. It may offer a novel, effective and technically simple technique for patients with no other therapeutic options. The results will be submitted to indexed medical journals and national and international meetings.Trial registration numberClinicalTrials.gov:NCT02798276, pre-results.

Altres ajuts: La Marató TV3 (201502 i 201516). This work has been developed in the context of AdvanceCat with the support of ACCIÓ (Catalonia Trade & Investment; Generalitat de Catalunya) under the Catalonian ERDF operational program (European Regional Development Fund) 2014-2020. FEB is sponsored by the "Researchers Stabilization Program" from the Spanish "Sistema Nacional de Salud" (SNS-ISCIII) and "Direcció d'Estratègia i Coordinació" Catalan Health Department (CES07/015). Undesired immune responses have drastically hampered outcomes after allogeneic organ transplantation and cell therapy, and also lead to inflammatory diseases and autoimmunity. Umbilical cord mesenchymal stem cells (UCMSCs) have powerful regenerative and immunomodulatory potential, and their secreted extracellular vesicles (EVs) are envisaged as a promising natural source of nanoparticles to increase outcomes in organ transplantation and control inflammatory diseases. However, poor EV preparations containing highly-abundant soluble proteins may mask genuine vesicular-associated functions and provide misleading data. Here, we used Size-Exclusion Chromatography (SEC) to successfully isolate EVs from UCMSCs-conditioned medium. These vesicles were defined as positive for CD9, CD63, CD73 and CD90, and their size and morphology characterized by NTA and cryo-EM. Their immunomodulatory potential was determined in polyclonal T cell proliferation assays, analysis of cytokine profiles and in the skewing of monocyte polarization. In sharp contrast to the non-EV containing fractions, to the complete conditioned medium and to ultracentrifuged pellet, SEC-purified EVs from UCMSCs inhibited T cell proliferation, resembling the effect of parental UCMSCs. Moreover, while SEC-EVs did not induce cytokine response, the non-EV fractions, conditioned medium and ultracentrifuged pellet promoted the secretion of pro-inflammatory cytokines by polyclonally stimulated T cells and supported Th17 polarization. In contrast, EVs did not induce monocyte polarization, but the non-EV fraction induced CD163 and CD206 expression and TNF-α production in monocytes. These findings increase the growing evidence confirming that EVs are an active component of MSC's paracrine immunosuppressive function and affirm their potential for therapeutics in nanomedicine. In addition, our results highlight the importance of well-purified and defined preparations of MSC-derived EVs to achieve the immunosuppressive effect.

Altres Ajuts: Fundació La MARATÓ de TV3 Grants 201516 and 201502; Fundació Daniel Bravo Andreu; Sociedad Española de Cardiología; Societat Catalana de Cardiologia; Generalitat de Catalunya Grant SGR 2014; and Acadèmia de Ciències Mèdiques i de la Salut de Catalunya i de Balears. Red de Terapia Celular Project RD12/0019/0029, Red de Investigación Cardiovascular Project RD12/0042/0047, (...) and cofounded by ISCIII-Sudirección General de Evaluación y el Fondo Europeo de Desarrollo Regional. Cardiac tissue engineering, which combines cells and biomaterials, is promising for limiting the sequelae of myocardial infarction (MI). We assessed myocardial function and scar evolution after implanting an engineered bioactive impedance graft (EBIG) in a swine MI model. The EBIG comprises a scaffold of decellularized human pericardium, green fluorescent protein-labeled porcine adipose tissue-derived progenitor cells (pATPCs), and a customized-design electrical impedance spectroscopy (EIS) monitoring system. Cardiac function was evaluated noninvasively by using magnetic resonance imaging (MRI). Scar healing was evaluated by using the EIS system within the implanted graft. Additionally, infarct size, fibrosis, and inflammation were explored by histopathology. Upon sacrifice 1 month after the intervention, MRI detected a significant improvement in left ventricular ejection fraction (7.5%64.9% vs. 1.4%63.7%; p = .038) and stroke volume (11.565.9 ml vs. 364.5 ml; p = .019) in EBIG-treated animals. Noninvasive EIS data analysis showed differences in both impedance magnitude ratio (20.02 6 0.04 per day vs. 20.48 6 0.07 per day; p = .002) and phase angle slope (20.18°60.24° per day vs.23.52°60.84° per day; p = .004) in EBIG compared with control animals. Moreover, in EBIG-treated animals, the infarct size was 48% smaller (3.4%60.6% vs. 6.5%61%; p = .015), less inflammation was found by means of CD25+ lymphocytes (0.65 6 0.12 vs. 1.26 6 0.2; p = .006), and a lower collagen I/III ratio was detected (0.4960.06 vs. 1.6660.5; p = .019). An EBIG composed of acellular pericardium refilled with pATPCs significantly reduced infarct size and improved cardiac function in a preclinical model of MI. Noninvasive EIS monitoring was useful for tracking differential scar healing in EBIG-treated animals, which was confirmed by less inflammation and altered collagen deposit.

A major challenge of cardiac tissue engineering is directing cells to establish the physiological structure and function of the myocardium being replaced. Our aim was to examine the effect of electrical stimulation on the cardiodifferentiation potential of cardiac adipose tissue-derived progenitor cells (cardiac ATDPCs). Three different electrical stimulation protocols were tested; the selected protocol consisted of 2 ms monophasic square-wave pulses of 50 mV/cm at 1 Hz over 14 days. Cardiac and subcutaneous ATDPCs were grown on biocompatible patterned surfaces. Cardiomyogenic differentiation was examined by real-time PCR and immunocytofluorescence. In cardiac ATDPCs, MEF2A and GATA-4 were significantly upregulated at day 14 after stimulation, while subcutaneous ATDPCs only exhibited increased Cx43 expression. In response to electrical stimulation, cardiac ATDPCs elongated, and both cardiac and subcutaneous ATDPCs became aligned following the linear surface pattern of the construct. Cardiac ATDPC length increased by 11.3%, while subcutaneous ATDPC length diminished by 11.2% (p = 0.013 and p = 0.030 vs unstimulated controls, respectively). Compared to controls, electrostimulated cells became aligned better to the patterned surfaces when the pattern was perpendicular to the electric field (89.71 ± 28.47o for cardiac ATDPCs and 92.15 ± 15.21o for subcutaneous ATDPCs). Electrical stimulation of cardiac ATDPCs caused changes in cell phenotype and genetic machinery, making them more suitable for cardiac regeneration approaches. Thus, it seems advisable to use electrical cell training before delivery as a cell suspension or within engineered tissue.

Advances in bioanalytical techniques have become crucial for both basic research and medical practice. One example, bioluminescence imaging (BLI), is based on the application of natural reactants with light-emitting capabilities (photoproteins and luciferases) isolated from a widespread group of organisms. The main challenges in cardiac regeneration remain unresolved, but a vast number of studies have harnessed BLI with the discovery of aequorin and green fluorescent proteins. First described in the luminous hydromedusan Aequorea victoria in the early 1960s, bioluminescent proteins have greatly contributed to the design and initiation of ongoing cell-based clinical trials on cardiovascular diseases. In conjunction with advances in reporter gene technology, BLI provides valuable information about the location and functional status of regenerative cells implanted into numerous animal models of disease. The purpose of this review was to present the great potential of BLI, among other existing imaging modalities, to refine effectiveness and underlying mechanisms of cardiac cell therapy. We recount the first discovery of natural primary compounds with light-emitting capabilities, and follow their applications to bioanalysis. We also illustrate insights and perspectives on BLI to illuminate current efforts in cardiac regeneration, where the future is bright.

Resumen La insuficiencia cardiaca es la etapa final de muchas enfermedades cardiovasculares, como el infarto agudo de miocardio, y sigue siendo uno de los retos mas atractivos para la medicina regenerativa debido a su alta incidencia y prevalencia. A lo largo de los ultimos 20 anos, la cardiomioplastia, basada en la administracion aislada de celulas con capacidad regenerativa, ha focalizado la mayoria de estudios que han perseguido regenerar el corazon. No obstante, aunque esta terapia se ha mostrado factible en el ambito clinico, el grado de regeneracion del miocardio infartado y de mejoria de la funcion cardiaca es muy limitado. Ante tal escenario ha emergido la ingenieria tisular cardiaca como una novedosa tecnologia basada en el uso de celulas con capacidad regenerativa, materiales biologicos y/o sinteticos, factores de crecimiento, diferenciacion y proangiogenicos, y sistemas de registro online para inducir la regeneracion de un organo o tejido danado. Un paso mas, segun algunos estudios pioneros realizados en animales, consiste en la generacion de corazones bioartificiales de novo descelularizandolos y preservando sus estructuras de soporte para posteriormente repoblarlos con nuevo tejido muscular contractil y vascular. Este nuevo abordaje comportaria, finalmente, el trasplante del corazon «reconstruido» restableciendo la funcion cardiaca del receptor.

There is compelling evidence that cardiovascular disorders arise and/or progress due mainly to endothelial dysfunction. Novel therapeutic strategies aim to generate new myocardial tissue using cells with regenerative potential, either alone or in combination with biomaterials, cytokines and advanced monitoring devices. Among the human adult progenitor cells used in such methods, those historically termed ‘endothelial progenitor cells' show promise for vascular growth and repair. Asahara et al. [Science 1997;275:964-967] initially described putative endothelial cell precursors in 1997. Subsequently, distinct cell populations termed endothelial colony-forming units-Hill, circulating angiogenic cells and endothelial colony-forming cells were identified that varied in terms of phenotype, vascular homeostasis contribution and purity. Notably, most of these cells are not genuine vascular precursor cells belonging to the endothelial lineage. This review provides a broad overview of the main properties of the endothelium, focusing on the basis governing its growth and repair. We discuss efforts to identify true vascular precursors, a matter of debate for the past 15 years, as well as recent methodological advances in identifying new hierarchies of more homogeneous, clonogenic and proliferative vascular endothelial-lineage precursors. Consideration of these issues provides insights that may help develop more effective therapies against human diseases that involve vascular deficits.

Recently, a new surgical technique to rescue infarcted myocardium (AGTP) has already tested (NCT01473433, AdiFLAP Trial). Here, we present the new minimally invasive AGTP (mi-AGTP) by thoracoscopy.