Layer-By-Layer Fabrication of Thicker and Larger Human Cardiac Muscle Patches for Cardiac Repair in Mice

The engineered myocardial tissues produced via most manufacturing techniques are typically just a few dozen micrometers thick, which is too thin for therapeutic applications in patients. Here, we used a modified layer-by-layer (LBL) fabrication protocol to generate thick human cardiac muscle patches (hCMPs) with thicknesses of ~3.75 mm. The LBL-hCMPs were composed of a layer of endothelial cells (ECs) sandwiched between two layers of cardiomyocytes (CMs): both cell populations were differentiated from the same human induced pluripotent stem cell line (hiPSCs) and suspended in a fibrin matrix, and the individual layers were sutured together, leaving channels that allowed the culture medium to access the internal cell layer. The LBL-hCMPs were cultured on a dynamic culture platform with electrical stimulation, and when compared to Control-hCMPs consisting of the same total number of hiPSC-ECs and -CMs suspended in a single layer of fibrin, hiPSC-CMs in the LBL-hCMPs were qualitatively more mature with significantly longer sarcomeres and expressed significantly higher levels of mRNA transcripts for proteins that participate in cardiomyocyte contractile activity and calcium handing. Apoptotic cells were also less common in LBL- than in Control-hCMPs. The thickness of fabricated LBL-hCMP gradually decreased to 0.8 mm by day 28 in dynamic culture. When the hCMP constructs were compared in a mouse model of myocardial infarction, the LBL-hCMPs were associated with significantly better measurements of engraftment, cardiac function, infarct size, hypertrophy, and vascularity. Collectively these observations indicate that our modified LBL fabrication protocol produced thicker hCMPs with no decline in cell viability, and that LBL-hCMPs were more potent than Control-hCMPs for promoting myocardial repair in mice.

Light-driven biological actuators to probe the rheology of 3D microtissues

The mechanical properties of biological tissues are key to the regulation of their physical integrity and function. Although the application of external loading or biochemical treatments allows to estimate these properties globally, it remains problematic to assess how such external stimuli compare with internal, cell-generated contractions. Here we engineered 3D microtissues composed of optogenetically-modified fibroblasts encapsulated within collagen. Using light to control the activity of RhoA, a major regulator of cellular contractility, we induced local mechanical perturbation within 3D fibrous microtissues, while tracking in real time microtissue stress and strain. We thus investigated the dynamic regulation of light-induced, local contractions and their spatio-temporal propagation in microtissues. By comparing the evolution of stresses and strains upon stimulation, we demonstrated the potential of our technique for quantifying tissue elasticity and viscosity, before examining the possibility of using light to map local anisotropies in mechanically heterogeneous microtissues. Altogether, our results open an avenue to non-destructively chart the rheology of 3D tissues in real time, using their own constituting cells as internal actuators.

Alterations in SLC4A2, SLC26A7 and SLC26A9 Drive Acid–Base Imbalance in Gastric Neuroendocrine Tumors and Uncover a Novel Mechanism for a Co-Occurring Polyautoimmune Scenario

Autoimmune polyendocrine syndrome (APS) is assumed to involve an immune system malfunction and entails several autoimmune diseases co-occurring in different tissues of the same patient; however, they are orphans of its accurate diagnosis, as its genetic basis and pathogenic mechanism are not understood. Our previous studies uncovered alterations in the ATPase H+/K+ Transporting Subunit Alpha (ATP4A) proton pump that triggered an internal cell acid–base imbalance, offering an autoimmune scenario for atrophic gastritis and gastric neuroendocrine tumors with secondary autoimmune pathologies. Here, we propose the genetic exploration of APS involving gastric disease to understand the underlying pathogenic mechanism of the polyautoimmune scenario. The whole exome sequencing (WES) study of five autoimmune thyrogastric families uncovered different pathogenic variants in SLC4A2, SLC26A7 and SLC26A9, which cotransport together with ATP4A. Exploratory in vitro studies suggested that the uncovered genes were involved in a pathogenic mechanism based on the alteration of the acid–base balance. Thus, we built a custom gene panel with 12 genes based on the suggested mechanism to evaluate a new series of 69 APS patients. In total, 64 filtered putatively damaging variants in the 12 genes of the panel were found in 54.17% of the studied patients and none of the healthy controls. Our studies reveal a constellation of solute carriers that co-express in the tissues affected with different autoimmune diseases, proposing a unique genetic origin for co-occurring pathologies. These results settle a new-fangled genetics-based mechanism for polyautoimmunity that explains not only gastric disease, but also thyrogastric pathology and disease co-occurrence in APS that are different from clinical incidental findings. This opens a new window leading to the prediction and diagnosis of co-occurring autoimmune diseases and clinical management of patients.

Analysis of the electrostatic field distribution in the hemispherical internal cell of radon monitors to estimate the collection efficiency of Po-218

The collection efficiency of the hemispherical internal cell of radon monitors depends on many factors, with the distribution of the electric field and the relative humidity of the air being particularly important. COMSOL is used to simulate an internal cell with a plastic upper surface. Simulation results show a relatively uniform gradient of the electric field. Assuming that the electric field felt by the positively charged Po-218 ions in the internal cell is a linear function of its radial coordinate, a mathematical model of the collection efficiency is proposed. From this model, we obtained the following: 1) under the same neutralization rate and potential, the electric field gradient has little effect on the collection efficiency; 2) under the same neutralization rate, the collection efficiency increases with the potential on the cell wall. If the neutralization rate is small, then the potential value for the maximum collection efficiency is also small. At a relative humidity of 6%–10%, the collection efficiency saturates for values of the electric potential on the cell wall larger than 5 kV; 3) under the same potential, a large neutralization rate corresponds to reduced collection efficiency. At high potential, the collection efficiency is relatively less affected by the neutralization rate. Higher collection efficiency can be achieved under high potential and low humidity conditions. This study provides a theoretical foundation to design the internal cell of radon monitor for improving the collection efficiency of Po-218.

An Insight into the Battery Degradation for a Proposal of a Battery Friendly Charging Technique

Li-ion batteries are widely used in electric vehicles because of their promising characteristics that meet high specific power and energy density requirements. The only setback is the capacity fading due to degradation in Li-ion batteries. The rate of capacity fade in Li-ion batteries in EVs vary based on the charging rate, changes in internal cell temperature and external ambient temperature, and user driving patterns. Since the Li-ion battery is electrochemical, determining the actual cause of degradation at a particular instant and constraining the rate is a big challenge. Further, the causes are related to parameters which are chemical, electrical and mechanical. In this work, the causes of degradation are studied by analysing the variation of parameters for multiple charge types and rates at different ambient temperatures. The analysis leads to developing a new universal charging scheme suitable to fast charge battery at different ambient temperatures appropriately and constrain battery degradation.

An Insight into the Battery Degradation and a Proposal for a Battery Friendly Charging Technique

Li-ion batteries are widely used in electric vehicles because of their promising characteristics that meet high specific power and energy density requirements. The only setback is the capacity fading due to degradation in Li-ion batteries. The rate of capacity fade in Li-ion batteries in EVs vary based on the charging rate, changes in internal cell temperature and external ambient temperature, and user driving patterns. Since the Li-ion battery is electrochemical, determining the actual cause of degradation at a particular instant and constraining the rate is a big challenge. Further, the causes are related to parameters which are chemical, electrical and mechanical. In this work, the causes of degradation are studied by analysing the variation of parameters for multiple charge types and rates at different ambient temperatures. The analysis leads to developing a new battery friendly charging scheme suitable to fast charge battery at different ambient temperatures appropriately and constrain battery degradation.

Analysis of Power Generation for Solar Photovoltaic Module with Various Internal Cell Spacing

Photovoltaic (PV) systems directly convert solar energy into electricity and researchers are taking into consideration the design of photovoltaic cell interconnections to form a photovoltaic module that maximizes solar irradiance. The purpose of this study is to evaluate the cell spacing effect of light diffusion on output power. In this work, the light absorption of solar PV cells in a module with three different cell spacings was studied. An optical engineering software program was used to analyze the reflecting light on the backsheet of the solar PV module towards the solar cell with varied internal cell spacing of 2 mm, 5 mm, and 8 mm. Then, assessments were performed under standard test conditions to investigate the power output of the PV modules. The results of the study show that the module with an internal cell spacing of 8 mm generated more power than 5 mm and 2 mm. Conversely, internal cell spacing from 2 mm to 5 mm revealed a greater increase of power output on the solar PV module compared to 5 mm to 8 mm. Furthermore, based on the simulation and experiment, internal cell spacing variation showed that the power output of a solar PV module can increase its potential to produce more power from the diffuse reflectance of light.

Methodology for Developing a Macro Finite Element Model of Lithium-Ion Pouch Cells for Predicting Mechanical Behaviour under Multiple Loading Conditions

To assist in light weighting of electric vehicles by improving the volumetric and gravimetric energy density and the structural performance of the battery pack, a modelling methodology based on a macro finite element model of a pouch cell has been developed. This model treats the core cell structure as a homogeneous orthotropic honeycomb block with the pouch material being defined as an orthotropic fabric with compressive stress elimination. The model considers five compression and bending load cases simultaneously and allows a level of element discretisation that is computationally efficient and appropriate for inclusion in full vehicle and sub-system simulations. The methodology is scalable in that it can be applied to a range of chemistries, external geometries and internal cell constructions. When considering stacks of cells, the model is predictive for both lateral compression and three-point bend, but further work is required to improve the confined compression response.

Persistent cell migration emerges from a coupling between protrusion dynamics and polarized trafficking

Migrating cells present a variety of paths, from random to highly directional ones. While random movement can be explained by basal intrinsic activity, persistent movement requires stable polarization. Here, we quantitatively address emergence of persistent migration in RPE1 cells over long timescales. By live-cell imaging and dynamic micropatterning, we demonstrate that the Nucleus-Golgi axis aligns with direction of migration leading to efficient cell movement. We show that polarized trafficking is directed towards protrusions with a 20 min delay, and that migration becomes random after disrupting internal cell organization. Eventually, we prove that localized optogenetic Cdc42 activation orients the Nucleus-Golgi axis. Our work suggests that polarized trafficking stabilizes the protrusive activity of the cell, while protrusive activity orients this polarity axis, leading to persistent cell migration. Using a minimal physical model, we show that this feedback is sufficient to recapitulate the quantitative properties of cell migration in the timescale of hours.