Myocardial remodeling is an inevitable risk factor for cardiac arrhythmias and can potentially be corrected with cell therapy. Although the generation of cardiac cells ex vivo is possible, specific approaches to cell replacement therapy remain unclear. On the one hand, adhesive myocyte cells must be viable and conjugated with the electromechanical syncytium of the recipient tissue, which is unattainable without an external scaffold substrate. On the other hand, the outer scaffold may hinder cell delivery, for example, making intramyocardial injection difficult. To resolve this contradiction, we developed molecular vehicles that combine a wrapped (rather than outer) polymer scaffold that is enveloped by the cell and provides excitability restoration (lost when cells were harvested) before engraftment. It also provides a coating with human fibronectin, which initiates the process of graft adhesion into the recipient tissue and can carry fluorescent markers for the external control of the non-invasive cell position. In this work, we used a type of scaffold that allowed us to use the advantages of a scaffold-free cell suspension for cell delivery. Fragmented nanofibers (0.85 µm ± 0.18 µm in diameter) with fluorescent labels were used, with solitary cells seeded on them. Cell implantation experiments were performed in vivo. The proposed molecular vehicles made it possible to establish rapid (30 min) electromechanical contact between excitable grafts and the recipient heart. Excitable grafts were visualized with optical mapping on a rat heart with Langendorff perfusion at a 0.72 ± 0.32 Hz heart rate. Thus, the pre-restored grafts’ excitability (with the help of a wrapped polymer scaffold) allowed rapid electromechanical coupling with the recipient tissue. This information could provide a basis for the reduction of engraftment arrhythmias in the first days after cell therapy.

In precision cardiology, virtual replicas (VRs) hold promise for predicting arrhythmias by leveraging patient-specific data and biophysics knowledge. A crucial first step is creating VRs of cardiac tissue based on retrospective patient data. However, VRs aim to replicate biopotential conduction directly, whereas only non-invasive methods are feasible for clinical use on real organs and tissues. This discrepancy challenges our understanding of VR applicability limits. This study aims to enhance the mathematical template of VR by developing an in vitro validation complement. We performed a frame-by-frame comparison of in vitro optical mapping of biopotential conduction with VR predictions. Patient-specific self-organized tissue samples from human induced pluripotent stem cell-derived cardiomyocytes (CMs) with diffuse fibrosis were utilized as VR prototypes. High-resolution optical mapping recordings (Δx = 117 ± 4 μm, Δt = 7.69 ms) and immunostaining were used to reproduce fibrotic samples of linear size 7.5 mm. We applied data-driven Bayesian optimization of the Cellular Potts model (CPM) to study wave propagation at the subcellular level. The modified CPM accurately reflected the “perinatal window” until the 20th day of differentiation, affecting CMs’ self-organization. The percolation threshold of virtual conductive pathways reached 0.26 (0.27 ± 0.03 of CMs in vitro), yielding a spatial correlation of amplitude maps with Pearson's coefficients of 0.83 ± 0.02. As a proof-of-concept, we demonstrated that CPM-enhanced VR could predict wavefront trajectories in optical mapping recordings, showing that approximating fibrosis distribution is crucial for improving VR prediction accuracy.

Cardiac arrhythmias are a major cause of cardiovascular mortality worldwide. Functional heterogeneity of cardiac tissue is an inevitable arrhythmogenic condition that may create nonlinear wave turbulence or reentry with subsequent arrhythmia initiation. The relation between propagation heterogeneity and the onset of reentry is of great theoretical and practical importance. Here, we present a conceptual representation of heterogeneous tissue expressed through alternating local and global tissue anisotropy with discreteness of membrane conductance. To contrast the influence of distributed heterogeneity, we investigated the interaction of a highfrequency wavetrain at a sharp anisotropy-symmetric obstacle. The revealed tendency of a heterogeneous system to form reentry was formalized into the single concept of a vulnerable frequency corridor that can be estimated experimentally. Using the joint in vitro–in silico approach, we defined an anomalous stable growth of a unidirectional block in the vicinity of an obstacle, depending on the direction of the anisotropy vector. This effect explains the limited applicability of homogeneous models to predicting the occurrence of primary reentry. Furthermore, computer simulations showed the special role played by other possible mechanisms of excitation, as ephaptic intercellular coupling, in the formation of a unidirectional

Introduction: Preservation of isolated hearts in cardioplegic solution at normothermia is a promising strategy for increasing the organ lifespan. Further development of this strategy and search for new products for solution delivery can help optimize the technology of donor heart transportation. Objective: The study was aimed to test the efficacy of device for normothermic constant perfusion flow with innovative lines for the cardioplegic solution delivery (CardioSystemPharma, Khimki, Moscow Region, Russia) according to the Langendorff mode in comparison with a system based on nodes and tubes of Radnoti Langendorff Constant Perfusion Flow Apparatus (ADInstruments, Colorado Springs, USA) in a series of experiments on rat hearts.Methods: The innovative cardioplegic delivery system (CardioSystemPharma) and traditional perfusion system (Radnoti) were compared in experiments on rat hearts at normothermia without cardioplegic solution and using the cardioplegic solution Normacor (Mosfarm, Moscow Region, Russia). The time until complete cardiac arrest and the changes in the NADH coenzyme fluorescence were recorded. After cardiac arrest, immunohistochemical staining of ventricular sections was performed to detect myocardial injury during perfusion.Results: It was shown that the innovative cardioplegic delivery system was not inferior to the traditional perfusion system (Radnoti) and even surpassed it in terms of the time duration for preservation of the isolated heart. It was found that the using of elevated glucose concentration during normothermic perfusion could significantly increase the organ lifespan. The efficacy of normothermic perfusion in the experiment was confirmed by monitoring cellular respiration through recording the NADH fluorescence.Conclusion: Testing showed that long-term preservation under normothermia with usage of an innovative delivery system of Russian manufacture is a promising replacement for Radnoti solution delivery system. Received 16 October 2024. Revised 23 November 2024. Accepted 10 December 2024. FundingThe study was carried out within the framework of the project “Innovative cardioplegic lines for cardiac surgery” under the Agreement on the provision of a grant from the Government of the Moscow Region in the fields of science, technology, engineering and innovation No. 12/04-24 dated 04/27/2024. Conflict of interestThe authors declare no conflict of interest. Contribution of the authorsConception and study design: V.A. Tsvelaya, S.D. Robustova, V.S. Kachan, M.S. Nosov, A.K. Berezhnoy, V.A. Kozlov, K.I. AgladzeData collection and analysis: S.D. Robustova, V.S. Kachan, D.V. Kononova, A.S. Yarovoy, M.A. Gusev, V.V. Litvinenko, V.D. Dzhabrailov, A.A. Brichagina, Z.V. Obrazumova, S.D. Chekalina, S.S. Bakumenko, E.A. Turchaninova, V.A. TsvelayaStatistic analysis: S.D. Robustova, V.S. Kachan, D.V. Kononova, A.S. Yarovoy, M.A. Gusev, V.V. Litvinenko, V.D. Dzhabrailov, A.A. Brichagina, Z.V. Obrazumova, S.D. Chekalina, S.S. Bakumenko, E.A. TurchaninovaDrafting the article: S.D. Robustova, V.S. Kachan, D.V. Kononova, Z.V. Obrazumova, E.A. Turchaninova, V.A. TsvelayaCritical revision of the article: S.D. Robustova, E.A. Turchaninova, V.A. Tsvelaya, K.I. AgladzeFinal approval of the version to be published: S.D. Robustova, V.S. Kachan, D.V. Kononova, A.S. Yarovoy, M.A. Gusev, V.V. Litvinenko, V.D. Dzhabrailov, A.A. Brichagina, Z.V. Obrazumova, S.D. Chekalina, E.A. Turchaninova, S.S. Bakumenko, M.S. Nosov, A.K. Berezhnoy, V.A. Kozlov, V.A. Tsvelaya, K.I. Agladze

Despite significant advances in diagnosis and treatment, atrial fibrillation remains a common cardiac arrhythmia affecting up to 2% of our world’s population. Atrial fibrillation results from complex dynamic interactions between risk factors and comorbidities that trigger a variety of atrial remodeling processes. Atrial remodeling increases the persistence of atrial fibrillation while contributing to disease progression. The variability of manifestations and the wide range of mechanisms involved in the initiation, maintenance, and progression of atrial fibrillation, as well as the associated adverse outcomes, make early identification of causative factors a major challenge for modern cardiology. Computer modeling over the past 60 years has opened new avenues for understanding mechanisms, risk prediction, and personalized therapy in the treatment of atrial fibrillation. As part of our modeling of patient-specific atrial structure from gadolinium-enhanced magnetic resonance imaging data, we investigated fibrosis as a substrate for the occurrence of re-entry from a purely electrophysiological perspective. We demonstrated the effects occurring on the patient-specific atrial structure in the case of normal electrophysiology and the electrophysiology obtained in long-term atrial fibrillation. In this work, we also compared the effects of the antiarrhythmic agent Verapamil on conduction by atrial models with different electrophysiology and fibrosis distribution.