Engineering and Assessing Cardiac Tissue Complexity

Cardiac tissue engineering is very much in a current focus of regenerative medicine research as it represents a promising strategy for cardiac disease modelling, cardiotoxicity testing and cardiovascular repair. Advances in this field over the last two decades have enabled the generation of human engineered cardiac tissue constructs with progressively increased functional capabilities. However, reproducing tissue-like properties is still a pending issue, as constructs generated to date remain immature relative to native adult heart. Moreover, there is a high degree of heterogeneity in the methodologies used to assess the functionality and cardiac maturation state of engineered cardiac tissue constructs, which further complicates the comparison of constructs generated in different ways. Here, we present an overview of the general approaches developed to generate functional cardiac tissues, discussing the different cell sources, biomaterials, and types of engineering strategies utilized to date. Moreover, we discuss the main functional assays used to evaluate the cardiac maturation state of the constructs, both at the cellular and the tissue levels. We trust that researchers interested in developing engineered cardiac tissue constructs will find the information reviewed here useful. Furthermore, we believe that providing a unified framework for comparison will further the development of human engineered cardiac tissue constructs displaying the specific properties best suited for each particular application

Quantitative volumetric Raman imaging of three dimensional cell cultures

The ability to simultaneously image multiple biomolecules in biologically relevant three-dimensional (3D) cell culture environments would contribute greatly to the understanding of complex cellular mechanisms and cell–material interactions. Here, we present a computational framework for label-free quantitative volumetric Raman imaging (qVRI). We apply qVRI to a selection of biological systems: human pluripotent stem cells with their cardiac derivatives, monocytes and monocyte-derived macrophages in conventional cell culture systems and mesenchymal stem cells inside biomimetic hydrogels that supplied a 3D cell culture environment. We demonstrate visualization and quantification of fine details in cell shape, cytoplasm, nucleus, lipid bodies and cytoskeletal structures in 3D with unprecedented biomolecular specificity for vibrational microspectroscopy

Factor analysis of linear alkylbenzene sulphonate (LAS) vertical distribution in coastal sediments of Cadiz Bay (southwest Spain)

El comportamiento del alquilbenceno lineal sulfonato (LAS) en los sistemas costeros no se conoce con precisión, y este conocimiento es aun menor si nos restringimos al compartimento sedimentario. En este trabajo se aplica un análisis factorial a los resultados obtenidos para distintas variables determinadas en tres estaciones, tanto en sedimento como en agua intersticial, con el objetivo de evaluar sus interrelaciones con el LAS. Las variables analizadas presentan dos modos principales de distribución con la profundidad en el compartimento sedimentario: lineal y/o exponencial. En casi todos los casos, el LAS se asocia a ambos modos de distribución, lo que indica que su concentración sufre una disminución con la profundidad, pero se produce de forma especialmente acusada en la capa superficial.Very little is known concerning the behaviour of linear alkylbenzene sulphonate (LAS) in coastal systems, and even less when we consider the sedimentary field only. In the present study, a factor analysis is applied to the results obtained for different variables at three stations, both in the sediment and in interstitial water, with the aim of evaluating their relationship with LAS. The variables analysed have two main types of distribution in the sedimentary area in relation to depth: linear and/or exponential. LAS is, in most cases, associated with both types of distribution, indicating that its concentration decreases with depth, especially in the surface layer.Instituto Español de Oceanografí

The fossil vertebrates from Somosaguas (Pozuelo, Madrid, Spain)

[ES] Dos yacimientos de vertebrados, situados en el Campus de Somosaguas de la Universidad Complutense (Pozuelo de Alarcón, Madrid), han proporcionado unos 600 restos identificables en estados de conservación muy variados, pertenecientes a unas veinte especies de tamaños muy diversos, desde mastodontes a musarañas. Su estudio permite fechar su edad en unos 14 m.a. y reconstruir un periodo árido en la cuenca de Madrid, ocupada durante el Mioceno medio por bosques y sabanas subtropicales con fuertes avenidas y sin ríos permanentes. En estos yacimientos se puede realizar una enseñanza práctica de la Paleontología de Vertebrados, para formación de estudiantes universitarios en el estudio y la gestión del Patrimonio Paleontológico.[EN] Two vertebrate fossil sites, situated in the Universidad Complutense Campus of Somosaguas, (Pozuelo de Alarcón, Madrid, Spain) have yielded about 600 identifiable rests in different preservation states, belonging to about twenty species of highly diverse sizes, from mastodons to shrews. Their study allows dating at about 14 m.y., and reconstructing an arid climate epoch in the Madrid basin during middle Miocene times, occupied by subtropical woodlands and savannahs with strong floods and without permanent rivers. These fossil sites allow practical teaching of Vertebrate Palaeontology, and preparing university students in the Palaeontological Heritage study and management.Hemos recibido financiación y personal respaldo del Rectorado de la Universidad Complutense de Madrid, de los Decanatos de la Facultad de Ciencias Políticas y Sociología y de la Facultad de Ciencias Geológicas, y del Departamento de Paleontología de esta Facultad. El Departamento de Paleobiología del Museo Nacional de Ciencias Naturales (CSIC) realiza su investigación en el marco del Convenio de Colaboración con la Comunidad de Madrid a través de la Dirección General de Patrimonio Histórico Artístico de la Consejería de Educación.Peer reviewe

A designer peptide as a template for growing Au nanoclusters

A peptide was designed to generate a sub-nanometric template that guides the growth of fluorescent gold nanoclusters. The peptide was endorsed with nucleating moieties and a three-dimensional structure that arrests the growth of ultrasmall nanoparticles. The nanoclusters are not cytotoxic and can be found in the cytosol of cells

Cells, Materials, and Fabrication Processes for Cardiac Tissue Engineering

Cardiovascular disease is the number one killer worldwide, with myocardial infarction (MI) responsible for approximately 1 in 6 deaths. The lack of endogenous regenerative capacity, added to the deleterious remodelling programme set into motion by myocardial necrosis, turns MI into a progressively debilitating disease, which current pharmacological therapy cannot halt. The advent of Regenerative Therapies over 2 decades ago kick-started a whole new scientific field whose aim was to prevent or even reverse the pathological processes of MI. As a highly dynamic organ, the heart displays a tight association between 3D structure and function, with the non-cellular components, mainly the cardiac extracellular matrix (ECM), playing both fundamental active and passive roles. Tissue engineering aims to reproduce this tissue architecture and function in order to fabricate replicas able to mimic or even substitute damaged organs. Recent advances in cell reprogramming and refinement of methods for additive manufacturing have played a critical role in the development of clinically relevant engineered cardiovascular tissues. This review focuses on the generation of human cardiac tissues for therapy, paying special attention to human pluripotent stem cells and their derivatives. We provide a perspective on progress in regenerative medicine from the early stages of cell therapy to the present day, as well as an overview of cellular processes, materials and fabrication strategies currently under investigation. Finally, we summarise current clinical applications and reflect on the most urgent needs and gaps to be filled for efficient translation to the clinical arena.This work was supported by funds from the ISCIII Red TERCEL RETIC RD16/0011/0005, PI 19/01350, ERANET II (Nanoreheart) and Gobierno de Navarra Departamento de Salud GNa8/2019, co-funded by FEDER funds, MINECO (Program RETOS Cardiomesh RTC-2016-4911-1), Gobierno de Navarra 0011-1383-2019-000006 and 0011-1383-2018-000011, and European Union's H2020 Program under grant agreement No. 874827 (BRAV(sic))

Transplantation of mesenchymal stem cells exerts a greater long-term effect than bone marrow mononuclear cells in a chronic myocardial infarction model in rat

The aim of this study is to assess the long-term effect of mesenchymal stem cells (MSC) transplantation in a rat model of chronic myocardial infarction (MI) in comparison with the effect of bone marrow mononuclear cells (BM-MNC) transplant. Five weeks after induction of MI, rats were allocated to receive intramyocardial injection of 106 GFP-expressing cells (BM-MNC or MSC) or medium as control. Heart function (echocardiography and 18F-FDG-microPET) and histological studies were performed 3 months after transplantation and cell fate was analyzed along the experiment (1 and 2 weeks and 1 and 3 months). The main findings of this study were that both BM-derived populations, BM-MNC and MSC, induced a long-lasting (3 months) improvement in LVEF (BM-MNC: 26.61 ± 2.01% to 46.61 ± 3.7%, p < 0.05; MSC: 27.5 ± 1.28% to 38.8 ± 3.2%, p < 0.05) but remarkably, only MSC improved tissue metabolism quantified by 18FFDG uptake (71.15 ± 1.27 to 76.31 ± 1.11, p < 0.01), which was thereby associated with a smaller infarct size and scar collagen content and also with a higher revascularization degree. Altogether, results show that MSC provides a long-term superior benefit than whole BM-MNC transplantation in a rat model of chronic MI

Plasticity and cardiovascular applications of multipotent adult progenitor cells

Cardiovascular disease is the leading cause of death worldwide, which has encouraged the search for new therapies that enable the treatment of patients in palliative and curative ways. In the past decade, the potential benefit of transplantation of cells that are able to substitute for the injured tissue has been studied with several cell populations, such as stem cells. Some of these cell populations, such as myoblasts and bone marrow cells, are already being used in clinical trials. The laboratory of CM Verfaillie has studied primitive progenitors, termed multipotent adult progenitor cells, which can be isolated from adult bone marrow. These cells can differentiate in vitro at the single-cell level into functional cells that belong to the three germ layers and contribute to most, if not all, somatic cell types after blastocyst injection. This remarkably broad differentiation potential makes this particular cell population a candidate for transplantation in tissues in need of regeneration. Here, we focus on the regenerative capacity of multipotent adult progenitor cells in several ischemic mouse models, such as acute and chronic myocardial infarction and limb ischemia

Generation of Self-Induced Myocardial Ischemia in Large-Sized Cardiac Spheroids without Alteration of Environmental Conditions Recreates Fibrotic Remodeling and Tissue Stiffening Revealed by Constriction Assays

Funding Information: This work has been supported by the European Union’s H2020 research and innovation programme under grant agreements No 829010 (PRIME H2020-FETOPEN-2018-2019-2020-01), 778354 (CISTEM H2020-MSCA-RISE-201), 874827 (BRAV∃), and 848109 (CRUCIAL); Instituto de Salud Carlos III cofinanced by European Regional Development Fund-FEDER “A way to make Europe” PI19/01350, PI21/00946 and CB16/11/00483; MCIN/AEI/10.13039/501100011033/ADVANCE (PID2022-139859OB-I00) and MCIN CARDIOPRINT (PLEC2021-008127); Gobierno de Navarra Proyectos Estratégicos IMPRIMED (0011-1411-2021-000096) and BIOHEART (0011-1411-2022-000071) and Gobierno de Navarra Proyectos Colaborativos BIOGEN (PC020-021-022). The regional Government of Aragon provided L.P. studentship. Publisher Copyright: © 2024 The Authors. Published by American Chemical Society.A combination of human-induced pluripotent stem cells (hiPSCs) and 3D microtissue culture techniques allows the generation of models that recapitulate the cardiac microenvironment for preclinical research of new treatments. In particular, spheroids represent the simplest approach to culture cells in 3D and generate gradients of cellular access to the media, mimicking the effects of an ischemic event. However, previous models required incubation under low oxygen conditions or deprived nutrient media to recreate ischemia. Here, we describe the generation of large spheroids (i.e., larger than 500 μm diameter) that self-induce an ischemic core. Spheroids were generated by coculture of cardiomyocytes derived from hiPSCs (hiPSC-CMs) and primary human cardiac fibroblast (hCF). In the proper medium, cells formed aggregates that generated an ischemic core 2 days after seeding. Spheroids also showed spontaneous cellular reorganization after 10 days, with hiPSC-CMs located at the center and surrounded by hCFs. This led to an increase in microtissue stiffness, characterized by the implementation of a constriction assay. All in all, these phenomena are hints of the fibrotic tissue remodeling secondary to a cardiac ischemic event, thus demonstrating the suitability of these spheroids for the modeling of human cardiac ischemia and its potential application for new treatments and drug research.publishersversionpublishe