Cardiovascular disease (CVD) remains the leading cause of death globally, with myocardial infarction (MI) being a major contributor. The current therapeutic approaches are limited in effectively regenerating damaged cardiac tissue. Up-to-date strategies for heart regeneration/reconstitution aim at cardiac remodeling through repairing the damaged tissue with an external cell source or by stimulating the existing cells to proliferate and repopulate the compromised area. Cell reprogramming is addressed to this challenge as a promising solution, converting fibroblasts and other cell types into functional cardiomyocytes, either by reverting cells to a pluripotent state or by directly switching cell lineage. Several strategies such as gene editing and the application of miRNA and small molecules have been explored for their potential to enhance cardiac regeneration. Those strategies take advantage of cell plasticity by introducing reprogramming factors that regress cell maturity in vitro, allowing for their later differentiation and thus endorsing cell transplantation, or promote in situ cell proliferation, leveraged by scaffolds embedded with pro-regenerative factors promoting efficient heart restoration. Despite notable advancements, important challenges persist, including low reprogramming efficiency, cell maturation limitations, and safety concerns in clinical applications. Nonetheless, integrating these innovative approaches offers a promising alternative for restoring cardiac function and reducing the dependency on full heart transplants.

Despite pharmacological-, technological- and medical- advances, cardiovascular diseases (CVDs) remain the main cause of death and disability in the world. This underscores the urgent need to better understand the early stages of these diseases for effective prevention, as well as to develop patient-specific pharmacological approaches (personalized medicine) and novel therapies. Traditional in vitro and in vivo models often fail to accurately mimic human physiology, limiting their translational potential. In this context, microphysiological systems (MPS) have emerged as advanced in vitro platforms that integrate key physiological features of cardiovascular tissues. This review summarizes the state-of-the-art advancements in in vitro models for studying CVDs, with a particular focus on emerging 3D cardiac and vascular models. These models serve as essential tools for disease modelling, drug development, and toxicity testing. Key parameters to consider when developing cardiovascular MPS are highlighted, along with a discussion of the advantages and challenges associated with each model system. Graphical Abstract

Cardiovascular diseases remain a growing concern worldwide. Hence, it is critical to understand cardiac development and disease in a relevant human-based in vitro model. Human cardiac organoids are an alternative approach to studying cardiogenesis, in the context of cell–cell communication, and disease etiology, using human induced pluripotent stem cells (hiPSCs). Extracellular vesicles (EVs) are nanosized particles harboring proteins, nucleic acids, and metabolites and are implicated in intercellular communication. Since cardiac development requires a complex interplay between several cell types, we hypothesize that EVs may mediate this communication. Here, we isolated EVs from hiPSC-derived cardiac organoids (cardEVs). LC-MS/MS was performed to analyze their protein cargo and compare it with those from a cardiomyocyte cell line (AC10 CM EVs) and from human heart explants of cadaveric donors (heEVs) using a bioinformatic approach. cardEVs share 48.9% of their proteins with heEVs, with important biological processes such as “Metabolism” and “Cardiac Function” highlighted in both proteomes. This overlap between the proteomes of cardEVs and heEVs suggests a molecular similarity between the two models. Therefore, we reiterate the importance of cardiac organoids as an excellent model for studying cardiac development and disease modeling, as well as to explore the complexity of intercellular communication.

After the publication of the review article