Despite significant advancements in in vitro cardiac modeling approaches, researchers still lack ... more Despite significant advancements in in vitro cardiac modeling approaches, researchers still lack the capacity to obtain in vitro measurements of a key indicator of cardiac function: contractility, or stroke volume under specific loading conditions—defined as the pressures to which the heart is subjected prior to and during contraction. This work puts forward a platform that creates this capability, by providing a means of dynamically controlling loading conditions in vitro. This dynamic tissue loading platform consists of a thin magnetoresponsive hydrogel cantilever on which 2D engineered myocardial tissue is cultured. Exposing the cantilever to an external magnetic field—generated by positioning magnets at a controlled distance from the cantilever—causes the hydrogel film to stretch, creating tissue load. Next, cell contraction is induced through electrical stimulation, and the force of the contraction is recorded, by measuring the cantilever's deflection. Force–length‐based me...
Extracellular vesicles (EVs) derived from various stem cell sources induce cardioprotective effec... more Extracellular vesicles (EVs) derived from various stem cell sources induce cardioprotective effects during ischemiareperfusion injury (IRI). These have been attributed mainly to the antiapoptotic, proangiogenic, microRNA (miRNA) cargo within the stem cell-derived EVs. However, the mechanisms of EV-mediated endothelial signaling to cardiomyocytes, as well as their therapeutic potential toward ischemic myocardial injury, are not clear. EV content beyond miRNA that may contribute to cardioprotection has not been fully illuminated. This study characterized the protein cargo of human vascular endothelial EVs (EEVs) to identify lead cardioactive proteins and assessed the effect of EEVs on human laminar cardiac tissues (hlCTs) exposed to IRI. We mapped the protein content of human vascular EEVs and identified proteins that were previously associated with cellular metabolism, redox state, and calcium handling, among other processes. Analysis of the protein landscape of human cardiomyocytes revealed corresponding modifications induced by EEV treatment. To assess their human-specific cardioprotection in vitro, we developed a human heart-on-a-chip IRI assay using human stem cell-derived, engineered cardiac tissues. We found that EEVs alleviated cardiac cell death as well as the loss in contractile capacity during and after simulated IRI in an uptake-and dose-dependent manner. Moreover, we found that EEVs increased the respiratory capacity of normoxic cardiomyocytes. These results suggest that vascular EEVs rescue hlCTs exposed to IRI possibly by supplementing injured myocytes with cargo that supports multiple metabolic and salvage pathways and therefore may serve as a multitargeted therapy for IRI.
Biomedical research has relied on animal studies and conventional cell cultures for decades. Rece... more Biomedical research has relied on animal studies and conventional cell cultures for decades. Recently, microphysiological systems (MPS), also known as organs-on-chips, that recapitulate the structure and function of native tissues in vitro, have emerged as a promising alternative 1. However, current MPS typically lack integrated sensors and their fabrication requires multi-step lithographic processes 2. Here, we introduce a facile route for fabricating a new class of instrumented cardiac microphysiological devices via multi-material 3D printing. Specifically, we designed six functional inks, based on piezo-resistive, high conductance, and biocompatible soft materials that enable integration of soft strain gauge sensors within micro-architectures that guide the self-assembly of physio-mimetic laminar cardiac tissues. We validated that these embedded sensors provide non-invasive, electronic readout of tissue contractile stresses, inside cell incubator environments. We further applied these devices to study drug responses, as well as the contractile development of human stem cell derived laminar cardiac tissues over four weeks. Users may view, print, copy, and download text and data-mine the content in such documents, for the purposes of academic research, subject always to the full Conditions of use:
In vitro disease models offer the ability to study specific systemic features in isolation to bet... more In vitro disease models offer the ability to study specific systemic features in isolation to better understand underlying mechanisms that lead to dysfunction. Here, we present a cardiac dysfunction model using angiotensin II (ANG II) to elicit pathological responses in a hearton-a-chip platform that recapitulates native laminar cardiac tissue structure. Our platform, composed of arrays of muscular thin films (MTF), allows for functional comparisons of healthy and diseased tissues by tracking film deflections resulting from contracting tissues. To test our model, we measured gene expression profiles, morphological remodeling, calcium transients, and contractile stress generation in response to ANG II exposure and compared against previous experimental and clinical results. We found that ANG II induced pathological gene expression profiles including over-expression of natriuretic peptide B, Rho GTPase 1, and T-type calcium channels. ANG II exposure also increased proarrhythmic early after depolarization events and significantly reduced peak systolic stresses. Although ANG II has been shown to induce structural remodeling, we control tissue architecture via microcontact printing, and show pathological genetic profiles and functional impairment precede significant morphological changes. We assert that our in vitro model is a useful tool for evaluating tissue health and can serve as a platform for studying disease mechanisms and identifying novel therapeutics.
Microphysiological systems and organs-on-chips promise to accelerate biomedical and pharmaceutica... more Microphysiological systems and organs-on-chips promise to accelerate biomedical and pharmaceutical research by providing accurate in vitro replicas of human tissue. Aside from addressing the physiological accuracy of the model tissues, there is a pressing need for improving the throughput of these platforms. To do so, scalable data acquisition strategies must be introduced. To this end, we here present an instrumented 24-well plate platform for higherthroughput studies of engineered human stem cell-derived cardiac muscle tissues that recapitulate the laminar structure of the native ventricle. In each well of the platform, an embedded flexible strain gauge provides continuous and non-invasive readout of the contractile stress and beat rate of an engineered cardiac tissue. The sensors are based on micro-cracked titanium-gold thin films, which ensure that the sensors are highly compliant and robust. We demonstrate the value of the platform for toxicology and drug-testing purposes by performing 12 complete dose-response studies of cardiac and cardiotoxic drugs. Additionally, we showcase the ability to couple the cardiac tissues with endothelial barriers. In these studies, which mimic the passage of drugs through the blood vessels to the musculature of the heart, we regulate the temporal onset of cardiac drug responses by modulating endothelial barrier permeability in vitro.
The development of scaffolding materials that recapitulate the cellular microenvironment and prov... more The development of scaffolding materials that recapitulate the cellular microenvironment and provide cells with physicochemical cues is crucial for successfully engineering functional tissues that can aid in repairing damaged organs. The use of gold nanoparticles for tissue engineering and regenerative medicine have raised great interest in recent years. In this mini review we will describe the shape dependent properties of gold nanoparticles, and their versatile use in creating tunable nanocomposite scaffolds, with improved mechanical and electrical properties for tissue engineering. We will further describe using gold nanoparticle-integrated scaffolds to achieve improved stem cells proliferation and differentiation. Finally, we will discuss the main challenges and prospects for clinical translation of gold nanoparticles-hybrid scaffolds.
Successfully engineering a functional, human, myocardial pump would represent a therapeutic alter... more Successfully engineering a functional, human, myocardial pump would represent a therapeutic alternative for the millions of patients with end-stage heart disease and provide an alternative to animal-based preclinical models. Although the field of cardiac tissue engineering has made tremendous advances, major challenges remain, which, if properly resolved, might allow the clinical implementation of engineered, functional, complex 3D structures in the future. In this Review, we provide an overview of state-of-the-art studies, challenges that have not yet been overcome and perspectives on cardiac tissue engineering. We begin with the most clinically relevant cell sources used in this field and discuss the use of topological, biophysical and metabolic stimuli to obtain mature phenotypes of cardiomyocytes, particularly in relation to organized cytoskeletal and contractile intracellular structures. We then move from the cellular level to engineering planar cardiac patches and discuss the need for proper vascularization and the main strategies for obtaining it. Finally, we provide an overview of several different approaches for the engineering of volumetric organs and organ parts — from whole-heart decellularization and recellularization to advanced 3D printing technologies. In this Review, Dvir and colleagues summarize and discuss the state-of-the-art technologies that enable the bioengineering of functional cardiac tissues and discuss current and future applications and limitations. The successful engineering of functional, human, myocardial tissues would represent a therapeutic alternative for the millions of patients with end-stage heart disease. The well-organized, characteristic structure of the heart across its multiple scales is crucial for its mechanical function; therefore, recapitulating these structure–function relationships in engineered myocardial tissues is necessary. Developing integrative maturation protocols and quantitative tools to assess the maturation of human induced pluripotent stem cell-derived cardiomyocytes is needed to produce cells with a mature phenotype that can successfully integrate with the host tissue. Although the engineering of anisotropic, thick, vascularized tissue patches has been demonstrated using various fabrication techniques, their clinical implementation is still hampered by their poor integration with host tissue. Fabricating 3D, volumetric, myocardial tissues could address the urgent need of patients with end-stage heart failure for heart donation and is therefore an active and evolving field of research.
ascularized human organ chips are microfluidic cell culture devices containing separate vascular ... more ascularized human organ chips are microfluidic cell culture devices containing separate vascular and parenchymal compartments lined by living human organ-specific cells. Such systems recapitulate the multicellular architecture, tissue-tissue interfaces and relevant physical microenvironments of key functional units of living organs while providing vascular perfusion in vitro 1,2. There is growing recognition that animal models do not effectively predict drug responses in humans 3-5 and a related increase in demand for in vitro human toxicity and efficacy testing. This has led to the pursuit of time-course analyses of human organ-chip models and fluidically linked, multi-organ, human-body-on-chip (HuBoC) systems that recapitulate organ-level functions to facilitate studies of the pharmacokinetics (PK) and pharmacodynamics (PD) of drugs in vitro 6-11. An automated experimental system that can meet these goals needs to be highly multifunctional and must ideally enable fluid handling and sample collection, perfusion of fluid through multiple linked microfluidic organ-chip devices and tissue imaging all within a controlled temperature, humidity and CO 2 environment. Custom assemblies using syringe pumps 12 , peristaltic pumps 13 , micropumps 11,14,15 and gravity feed 10,16,17 , including several commercial systems 18,19 , can be used to perfuse and link microfluidic tissue and organ culture inside incubators; however, these systems do not provide the features necessary for complex HuBoC experimentation. In particular, these systems offer limited ability to sample the different biological compartments and lack the capacity to reconfigure the system so that the same system can be used for multiple
Conducting polymer hydrogels (CPHs) emerge as excellent functional materials, as they harness the... more Conducting polymer hydrogels (CPHs) emerge as excellent functional materials, as they harness the advantages of conducting polymers with the mechanical properties and continuous 3D nanostructures of hydrogels. This bicomponent organization results in soft, all-organic, conducting micro-/nanostructures with multifarious material applications. However, the application of CPHs as functional materials for biomedical applications is currently limited due to the necessity to combine the features of biocompatibility, self-healing, and fine-tuning of the mechanical properties. To overcome this issue, we choose to combine a protected dipeptide as the supramolecular gelator, owing to its intrinsic biocompatibility and excellent gelation ability, with the conductive polymer polyaniline (PAni), which was polymerized in situ. Thus, a twocomponent, all-organic, conducting hydrogel was formed. Spectroscopic evidence reveals the formation of the emeraldine salt form of PAni by intrinsic doping. The composite hydrogel is mechanically rigid with a very high storage modulus (G′) value of ~2 MPa, and the rigidity was tuned by changing the peptide concentration. The hydrogel exhibits ohmic conductivity, pressure sensitivity, and, importantly, self-healing features. By virtue of its self-healing property, the polymeric nonmetallic hydrogel can reinstate its intrinsic conductivity when two of its macroscopically separated blocks are rejoined. High cell viability of cardiomyocytes grown on the composite hydrogel demonstrates its noncytotoxicity. These combined attributes of the hydrogel allowed its utilization for dynamic range pressure sensing and as a conductive interface for iD ORCID
The development of scaffolding materials that recapitulate the cellular microenvironment and prov... more The development of scaffolding materials that recapitulate the cellular microenvironment and provide cells with physicochemical cues is crucial for successfully engineering functional tissues that can aid in repairing damaged organs. The use of gold nanoparticles for tissue engineering and regenerative medicine have raised great interest in recent years. In this mini review we will describe the shape dependent properties of gold nanoparticles, and their versatile use in creating tunable nanocomposite scaffolds, with improved mechanical and electrical properties for tissue engineering. We will further describe using gold nanoparticle-integrated scaffolds to achieve improved stem cells proliferation and differentiation. Finally, we will discuss the main challenges and prospects for clinical translation of gold nanoparticles-hybrid scaffolds.
Here we describe of an Interrogator instrument that uses liquid-handling robotics, a custom softw... more Here we describe of an Interrogator instrument that uses liquid-handling robotics, a custom software package, and an integrated mobile microscope to enable automated culture, perfusion, medium addition, fluidic linking, sample collection, and in situ microscopic imaging of up to 10 Organ Chips inside a standard tissue culture incubator. The automated Interrogator platform maintained the viability and organ-specific functions of 8 different vascularized, 2-channel, Organ Chips (intestine, liver, kidney, heart, lung, skin, blood-brain barrier (BBB), and brain) for 3 weeks in culture when fluidically coupled through their endothelium-lined vascular channels using a common blood substitute medium. When an inulin tracer was perfused through the multi-organ Human Body-on-Chips (HuBoC) fluidic network, quantitative distributions of this tracer could be accurately predicted using a physiologically-based multi-compartmental reduced order (MCRO) in silico model of the experimental system deri...
Here we demonstrate that microfluidic cell culture devices, known as Organs-on-a-Chips can be fab... more Here we demonstrate that microfluidic cell culture devices, known as Organs-on-a-Chips can be fabricated with multifunctional, real-time, sensing capabilities by integrating both multi-electrode arrays (MEAs) and electrodes for transepithelial electrical resistance (TEER) measurements into the chips during their fabrication. To prove proof-of-concept, simultaneous measurements of cellular electrical activity and tissue barrier function were carried out in a dual channel, endothelialized, heart-on-a-chip device containing human cardiomyocytes and a channel-separating porous membrane covered with a primary human endothelial cell monolayer. These studies confirmed that the TEER-MEA chip can be used to simultaneously detect dynamic alterations of vascular permeability and cardiac function in the same chip when challenged with the inflammatory stimulus tumor necrosis factor alpha (TNF-α) or the cardiac targeting drug isoproterenol. Thus, this Organ Chip with integrated sensing capability...
Intracellular recording of action potentials is an essential mean for studying disease mechanisms... more Intracellular recording of action potentials is an essential mean for studying disease mechanisms, and for electrophysiological studies, particularly in excitable cells as cardiomyocytes or neurons. Current strategies to obtain intracellular recordings include three-dimensional (3D) nanoelectrodes that can effectively penetrate the cell membrane and achieve high-quality intracellular recordings in a minimally invasive manner, or transient electroporation of the membrane that can yield temporary intracellular access. However, the former strategy requires a complicated and costly fabrication process, and the latter strategy suffers from high dependency on the method of application of electroporation, yielding inconsistent, suboptimal recordings. These factors hinder the high throughput use of these strategies in electrophysiological studies. In this work, we propose an advanced cell-based biosensing platform that relies on electroporation to produce consistent, high-quality intracellular recordings. The suggested universal system can be integrated with any electrode array, and it enables tunable electroporation with controllable pulse parameters, while the recorded potentials can be analyzed in real time to provide instantaneous feedback on the electroporation effectiveness. This integrated system enables the user to perform electroporation, record and assess the obtained signals in a facile manner, to ultimately achieve stable, reliable, intracellular recording. Moreover, the proposed platform relies on microelectrode arrays which are suited for large-scale production, and additional modules that are low-cost. Using this platform, we demonstrate the tuning of electroporation pulse width, pulse number, and amplitude, to achieve effective electroporation and high-quality intracellular recordings. This integrated platform has the potential to enable larger scale, repeatable, convenient, and low-cost electrophysiological studies.
Microphysiological systems and organs-on-chips promise to accelerate biomedical and pharmaceutica... more Microphysiological systems and organs-on-chips promise to accelerate biomedical and pharmaceutical research by providing accurate in vitro replicas of human tissue. Aside from addressing the physiological accuracy of the model tissues, there is a pressing need for improving the throughput of these platforms. To do so, scalable data acquisition strategies must be introduced. To this end, we here present an instrumented 24-well plate platform for higher-throughput studies of engineered human stem cell-derived cardiac muscle tissues that recapitulate the laminar structure of the native ventricle. In each well of the platform, an embedded flexible strain gauge provides continuous and non-invasive readout of the contractile stress and beat rate of an engineered cardiac tissue. The sensors are based on micro-cracked titanium-gold thin films, which ensure that the sensors are highly compliant and robust. We demonstrate the value of the platform for toxicology and drug-testing purposes by p...
In vitro disease models offer the ability to study specific systemic features in isolation to bet... more In vitro disease models offer the ability to study specific systemic features in isolation to bet-ter understand underlying mechanisms that lead to dysfunction. Here, we present a cardiac dysfunction model using angiotensin II (ANG II) to elicit pathological responses in a heart-on-a-chip platform that recapitulates native laminar cardiac tissue structure. Our platform, composed of arrays of muscular thin films (MTF), allows for functional comparisons of healthy and diseased tissues by tracking film deflections resulting from contracting tissues. To test our model, we measured gene expression profiles, morphological remodeling, cal-cium transients, and contractile stress generation in response to ANG II exposure and com-pared against previous experimental and clinical results. We found that ANG II induced pathological gene expression profiles including over-expression of natriuretic peptide B, Rho GTPase 1, and T-type calcium channels. ANG II exposure also increased proarrhyth-mic e...
Biomedical research has relied on animal studies and conventional cell cultures for decades. Rece... more Biomedical research has relied on animal studies and conventional cell cultures for decades. Recently, microphysiological systems (MPS), also known as organs-on-chips, that recapitulate the structure and function of native tissues in vitro, have emerged as a promising alternative. However, current MPS typically lack integrated sensors and their fabrication requires multi-step lithographic processes. Here, we introduce a facile route for fabricating anewclass of instrumented cardiacmicrophysiological devicesviamultimaterial three-dimensional (3D) printing. Specifically, we designed six functional inks, based on piezo-resistive, high-conductance, and biocompatible soft materials that enable integration of soft strain gauge sensors within micro-architectures that guide the self-assembly of physio-mimetic laminar cardiac tissues. We validated that these embedded sensors provide non-invasive, electronic readouts of tissue contractile stresses inside cell incubator environments. We furthe...
Extracellular vesicles (EVs) derived from various stem cell sources induce cardioprotective effec... more Extracellular vesicles (EVs) derived from various stem cell sources induce cardioprotective effects during ischemia-reperfusion injury (IRI). These have been attributed mainly to the antiapoptotic, proangiogenic, microRNA (miRNA) cargo within the stem cell–derived EVs. However, the mechanisms of EV-mediated endothelial signaling to cardiomyocytes, as well as their therapeutic potential toward ischemic myocardial injury, are not clear. EV content beyond miRNA that may contribute to cardioprotection has not been fully illuminated. This study characterized the protein cargo of human vascular endothelial EVs (EEVs) to identify lead cardioactive proteins and assessed the effect of EEVs on human laminar cardiac tissues (hlCTs) exposed to IRI. We mapped the protein content of human vascular EEVs and identified proteins that were previously associated with cellular metabolism, redox state, and calcium handling, among other processes. Analysis of the protein landscape of human cardiomyocytes...
Biomedical research has relied on animal studies and conventional cell cultures for decades. Rece... more Biomedical research has relied on animal studies and conventional cell cultures for decades. Recently, microphysiological systems (MPS), also known as organs-on-chips, that recapitulate the structure and function of native tissues in vitro, have emerged as a promising alternative. However, current MPS typically lack integrated sensors and their fabrication requires multi-step lithographic processes. Here, we introduce a facile route for fabricating a new class of instrumented cardiac microphysiological devices via multimaterial three-dimensional (3D) printing. Specifically, we designed six functional inks, based on piezo-resistive, high-conductance, and biocompatible soft materials that enable integration of soft strain gauge sensors within micro-architectures that guide the self-assembly of physio-mimetic laminar cardiac tissues. We validated that these embedded sensors provide non-invasive, electronic readouts of tissue contractile stresses inside cell incubator environments. We f...
Despite significant advancements in in vitro cardiac modeling approaches, researchers still lack ... more Despite significant advancements in in vitro cardiac modeling approaches, researchers still lack the capacity to obtain in vitro measurements of a key indicator of cardiac function: contractility, or stroke volume under specific loading conditions—defined as the pressures to which the heart is subjected prior to and during contraction. This work puts forward a platform that creates this capability, by providing a means of dynamically controlling loading conditions in vitro. This dynamic tissue loading platform consists of a thin magnetoresponsive hydrogel cantilever on which 2D engineered myocardial tissue is cultured. Exposing the cantilever to an external magnetic field—generated by positioning magnets at a controlled distance from the cantilever—causes the hydrogel film to stretch, creating tissue load. Next, cell contraction is induced through electrical stimulation, and the force of the contraction is recorded, by measuring the cantilever's deflection. Force–length‐based me...
Extracellular vesicles (EVs) derived from various stem cell sources induce cardioprotective effec... more Extracellular vesicles (EVs) derived from various stem cell sources induce cardioprotective effects during ischemiareperfusion injury (IRI). These have been attributed mainly to the antiapoptotic, proangiogenic, microRNA (miRNA) cargo within the stem cell-derived EVs. However, the mechanisms of EV-mediated endothelial signaling to cardiomyocytes, as well as their therapeutic potential toward ischemic myocardial injury, are not clear. EV content beyond miRNA that may contribute to cardioprotection has not been fully illuminated. This study characterized the protein cargo of human vascular endothelial EVs (EEVs) to identify lead cardioactive proteins and assessed the effect of EEVs on human laminar cardiac tissues (hlCTs) exposed to IRI. We mapped the protein content of human vascular EEVs and identified proteins that were previously associated with cellular metabolism, redox state, and calcium handling, among other processes. Analysis of the protein landscape of human cardiomyocytes revealed corresponding modifications induced by EEV treatment. To assess their human-specific cardioprotection in vitro, we developed a human heart-on-a-chip IRI assay using human stem cell-derived, engineered cardiac tissues. We found that EEVs alleviated cardiac cell death as well as the loss in contractile capacity during and after simulated IRI in an uptake-and dose-dependent manner. Moreover, we found that EEVs increased the respiratory capacity of normoxic cardiomyocytes. These results suggest that vascular EEVs rescue hlCTs exposed to IRI possibly by supplementing injured myocytes with cargo that supports multiple metabolic and salvage pathways and therefore may serve as a multitargeted therapy for IRI.
Biomedical research has relied on animal studies and conventional cell cultures for decades. Rece... more Biomedical research has relied on animal studies and conventional cell cultures for decades. Recently, microphysiological systems (MPS), also known as organs-on-chips, that recapitulate the structure and function of native tissues in vitro, have emerged as a promising alternative 1. However, current MPS typically lack integrated sensors and their fabrication requires multi-step lithographic processes 2. Here, we introduce a facile route for fabricating a new class of instrumented cardiac microphysiological devices via multi-material 3D printing. Specifically, we designed six functional inks, based on piezo-resistive, high conductance, and biocompatible soft materials that enable integration of soft strain gauge sensors within micro-architectures that guide the self-assembly of physio-mimetic laminar cardiac tissues. We validated that these embedded sensors provide non-invasive, electronic readout of tissue contractile stresses, inside cell incubator environments. We further applied these devices to study drug responses, as well as the contractile development of human stem cell derived laminar cardiac tissues over four weeks. Users may view, print, copy, and download text and data-mine the content in such documents, for the purposes of academic research, subject always to the full Conditions of use:
In vitro disease models offer the ability to study specific systemic features in isolation to bet... more In vitro disease models offer the ability to study specific systemic features in isolation to better understand underlying mechanisms that lead to dysfunction. Here, we present a cardiac dysfunction model using angiotensin II (ANG II) to elicit pathological responses in a hearton-a-chip platform that recapitulates native laminar cardiac tissue structure. Our platform, composed of arrays of muscular thin films (MTF), allows for functional comparisons of healthy and diseased tissues by tracking film deflections resulting from contracting tissues. To test our model, we measured gene expression profiles, morphological remodeling, calcium transients, and contractile stress generation in response to ANG II exposure and compared against previous experimental and clinical results. We found that ANG II induced pathological gene expression profiles including over-expression of natriuretic peptide B, Rho GTPase 1, and T-type calcium channels. ANG II exposure also increased proarrhythmic early after depolarization events and significantly reduced peak systolic stresses. Although ANG II has been shown to induce structural remodeling, we control tissue architecture via microcontact printing, and show pathological genetic profiles and functional impairment precede significant morphological changes. We assert that our in vitro model is a useful tool for evaluating tissue health and can serve as a platform for studying disease mechanisms and identifying novel therapeutics.
Microphysiological systems and organs-on-chips promise to accelerate biomedical and pharmaceutica... more Microphysiological systems and organs-on-chips promise to accelerate biomedical and pharmaceutical research by providing accurate in vitro replicas of human tissue. Aside from addressing the physiological accuracy of the model tissues, there is a pressing need for improving the throughput of these platforms. To do so, scalable data acquisition strategies must be introduced. To this end, we here present an instrumented 24-well plate platform for higherthroughput studies of engineered human stem cell-derived cardiac muscle tissues that recapitulate the laminar structure of the native ventricle. In each well of the platform, an embedded flexible strain gauge provides continuous and non-invasive readout of the contractile stress and beat rate of an engineered cardiac tissue. The sensors are based on micro-cracked titanium-gold thin films, which ensure that the sensors are highly compliant and robust. We demonstrate the value of the platform for toxicology and drug-testing purposes by performing 12 complete dose-response studies of cardiac and cardiotoxic drugs. Additionally, we showcase the ability to couple the cardiac tissues with endothelial barriers. In these studies, which mimic the passage of drugs through the blood vessels to the musculature of the heart, we regulate the temporal onset of cardiac drug responses by modulating endothelial barrier permeability in vitro.
The development of scaffolding materials that recapitulate the cellular microenvironment and prov... more The development of scaffolding materials that recapitulate the cellular microenvironment and provide cells with physicochemical cues is crucial for successfully engineering functional tissues that can aid in repairing damaged organs. The use of gold nanoparticles for tissue engineering and regenerative medicine have raised great interest in recent years. In this mini review we will describe the shape dependent properties of gold nanoparticles, and their versatile use in creating tunable nanocomposite scaffolds, with improved mechanical and electrical properties for tissue engineering. We will further describe using gold nanoparticle-integrated scaffolds to achieve improved stem cells proliferation and differentiation. Finally, we will discuss the main challenges and prospects for clinical translation of gold nanoparticles-hybrid scaffolds.
Successfully engineering a functional, human, myocardial pump would represent a therapeutic alter... more Successfully engineering a functional, human, myocardial pump would represent a therapeutic alternative for the millions of patients with end-stage heart disease and provide an alternative to animal-based preclinical models. Although the field of cardiac tissue engineering has made tremendous advances, major challenges remain, which, if properly resolved, might allow the clinical implementation of engineered, functional, complex 3D structures in the future. In this Review, we provide an overview of state-of-the-art studies, challenges that have not yet been overcome and perspectives on cardiac tissue engineering. We begin with the most clinically relevant cell sources used in this field and discuss the use of topological, biophysical and metabolic stimuli to obtain mature phenotypes of cardiomyocytes, particularly in relation to organized cytoskeletal and contractile intracellular structures. We then move from the cellular level to engineering planar cardiac patches and discuss the need for proper vascularization and the main strategies for obtaining it. Finally, we provide an overview of several different approaches for the engineering of volumetric organs and organ parts — from whole-heart decellularization and recellularization to advanced 3D printing technologies. In this Review, Dvir and colleagues summarize and discuss the state-of-the-art technologies that enable the bioengineering of functional cardiac tissues and discuss current and future applications and limitations. The successful engineering of functional, human, myocardial tissues would represent a therapeutic alternative for the millions of patients with end-stage heart disease. The well-organized, characteristic structure of the heart across its multiple scales is crucial for its mechanical function; therefore, recapitulating these structure–function relationships in engineered myocardial tissues is necessary. Developing integrative maturation protocols and quantitative tools to assess the maturation of human induced pluripotent stem cell-derived cardiomyocytes is needed to produce cells with a mature phenotype that can successfully integrate with the host tissue. Although the engineering of anisotropic, thick, vascularized tissue patches has been demonstrated using various fabrication techniques, their clinical implementation is still hampered by their poor integration with host tissue. Fabricating 3D, volumetric, myocardial tissues could address the urgent need of patients with end-stage heart failure for heart donation and is therefore an active and evolving field of research.
ascularized human organ chips are microfluidic cell culture devices containing separate vascular ... more ascularized human organ chips are microfluidic cell culture devices containing separate vascular and parenchymal compartments lined by living human organ-specific cells. Such systems recapitulate the multicellular architecture, tissue-tissue interfaces and relevant physical microenvironments of key functional units of living organs while providing vascular perfusion in vitro 1,2. There is growing recognition that animal models do not effectively predict drug responses in humans 3-5 and a related increase in demand for in vitro human toxicity and efficacy testing. This has led to the pursuit of time-course analyses of human organ-chip models and fluidically linked, multi-organ, human-body-on-chip (HuBoC) systems that recapitulate organ-level functions to facilitate studies of the pharmacokinetics (PK) and pharmacodynamics (PD) of drugs in vitro 6-11. An automated experimental system that can meet these goals needs to be highly multifunctional and must ideally enable fluid handling and sample collection, perfusion of fluid through multiple linked microfluidic organ-chip devices and tissue imaging all within a controlled temperature, humidity and CO 2 environment. Custom assemblies using syringe pumps 12 , peristaltic pumps 13 , micropumps 11,14,15 and gravity feed 10,16,17 , including several commercial systems 18,19 , can be used to perfuse and link microfluidic tissue and organ culture inside incubators; however, these systems do not provide the features necessary for complex HuBoC experimentation. In particular, these systems offer limited ability to sample the different biological compartments and lack the capacity to reconfigure the system so that the same system can be used for multiple
Conducting polymer hydrogels (CPHs) emerge as excellent functional materials, as they harness the... more Conducting polymer hydrogels (CPHs) emerge as excellent functional materials, as they harness the advantages of conducting polymers with the mechanical properties and continuous 3D nanostructures of hydrogels. This bicomponent organization results in soft, all-organic, conducting micro-/nanostructures with multifarious material applications. However, the application of CPHs as functional materials for biomedical applications is currently limited due to the necessity to combine the features of biocompatibility, self-healing, and fine-tuning of the mechanical properties. To overcome this issue, we choose to combine a protected dipeptide as the supramolecular gelator, owing to its intrinsic biocompatibility and excellent gelation ability, with the conductive polymer polyaniline (PAni), which was polymerized in situ. Thus, a twocomponent, all-organic, conducting hydrogel was formed. Spectroscopic evidence reveals the formation of the emeraldine salt form of PAni by intrinsic doping. The composite hydrogel is mechanically rigid with a very high storage modulus (G′) value of ~2 MPa, and the rigidity was tuned by changing the peptide concentration. The hydrogel exhibits ohmic conductivity, pressure sensitivity, and, importantly, self-healing features. By virtue of its self-healing property, the polymeric nonmetallic hydrogel can reinstate its intrinsic conductivity when two of its macroscopically separated blocks are rejoined. High cell viability of cardiomyocytes grown on the composite hydrogel demonstrates its noncytotoxicity. These combined attributes of the hydrogel allowed its utilization for dynamic range pressure sensing and as a conductive interface for iD ORCID
The development of scaffolding materials that recapitulate the cellular microenvironment and prov... more The development of scaffolding materials that recapitulate the cellular microenvironment and provide cells with physicochemical cues is crucial for successfully engineering functional tissues that can aid in repairing damaged organs. The use of gold nanoparticles for tissue engineering and regenerative medicine have raised great interest in recent years. In this mini review we will describe the shape dependent properties of gold nanoparticles, and their versatile use in creating tunable nanocomposite scaffolds, with improved mechanical and electrical properties for tissue engineering. We will further describe using gold nanoparticle-integrated scaffolds to achieve improved stem cells proliferation and differentiation. Finally, we will discuss the main challenges and prospects for clinical translation of gold nanoparticles-hybrid scaffolds.
Here we describe of an Interrogator instrument that uses liquid-handling robotics, a custom softw... more Here we describe of an Interrogator instrument that uses liquid-handling robotics, a custom software package, and an integrated mobile microscope to enable automated culture, perfusion, medium addition, fluidic linking, sample collection, and in situ microscopic imaging of up to 10 Organ Chips inside a standard tissue culture incubator. The automated Interrogator platform maintained the viability and organ-specific functions of 8 different vascularized, 2-channel, Organ Chips (intestine, liver, kidney, heart, lung, skin, blood-brain barrier (BBB), and brain) for 3 weeks in culture when fluidically coupled through their endothelium-lined vascular channels using a common blood substitute medium. When an inulin tracer was perfused through the multi-organ Human Body-on-Chips (HuBoC) fluidic network, quantitative distributions of this tracer could be accurately predicted using a physiologically-based multi-compartmental reduced order (MCRO) in silico model of the experimental system deri...
Here we demonstrate that microfluidic cell culture devices, known as Organs-on-a-Chips can be fab... more Here we demonstrate that microfluidic cell culture devices, known as Organs-on-a-Chips can be fabricated with multifunctional, real-time, sensing capabilities by integrating both multi-electrode arrays (MEAs) and electrodes for transepithelial electrical resistance (TEER) measurements into the chips during their fabrication. To prove proof-of-concept, simultaneous measurements of cellular electrical activity and tissue barrier function were carried out in a dual channel, endothelialized, heart-on-a-chip device containing human cardiomyocytes and a channel-separating porous membrane covered with a primary human endothelial cell monolayer. These studies confirmed that the TEER-MEA chip can be used to simultaneously detect dynamic alterations of vascular permeability and cardiac function in the same chip when challenged with the inflammatory stimulus tumor necrosis factor alpha (TNF-α) or the cardiac targeting drug isoproterenol. Thus, this Organ Chip with integrated sensing capability...
Intracellular recording of action potentials is an essential mean for studying disease mechanisms... more Intracellular recording of action potentials is an essential mean for studying disease mechanisms, and for electrophysiological studies, particularly in excitable cells as cardiomyocytes or neurons. Current strategies to obtain intracellular recordings include three-dimensional (3D) nanoelectrodes that can effectively penetrate the cell membrane and achieve high-quality intracellular recordings in a minimally invasive manner, or transient electroporation of the membrane that can yield temporary intracellular access. However, the former strategy requires a complicated and costly fabrication process, and the latter strategy suffers from high dependency on the method of application of electroporation, yielding inconsistent, suboptimal recordings. These factors hinder the high throughput use of these strategies in electrophysiological studies. In this work, we propose an advanced cell-based biosensing platform that relies on electroporation to produce consistent, high-quality intracellular recordings. The suggested universal system can be integrated with any electrode array, and it enables tunable electroporation with controllable pulse parameters, while the recorded potentials can be analyzed in real time to provide instantaneous feedback on the electroporation effectiveness. This integrated system enables the user to perform electroporation, record and assess the obtained signals in a facile manner, to ultimately achieve stable, reliable, intracellular recording. Moreover, the proposed platform relies on microelectrode arrays which are suited for large-scale production, and additional modules that are low-cost. Using this platform, we demonstrate the tuning of electroporation pulse width, pulse number, and amplitude, to achieve effective electroporation and high-quality intracellular recordings. This integrated platform has the potential to enable larger scale, repeatable, convenient, and low-cost electrophysiological studies.
Microphysiological systems and organs-on-chips promise to accelerate biomedical and pharmaceutica... more Microphysiological systems and organs-on-chips promise to accelerate biomedical and pharmaceutical research by providing accurate in vitro replicas of human tissue. Aside from addressing the physiological accuracy of the model tissues, there is a pressing need for improving the throughput of these platforms. To do so, scalable data acquisition strategies must be introduced. To this end, we here present an instrumented 24-well plate platform for higher-throughput studies of engineered human stem cell-derived cardiac muscle tissues that recapitulate the laminar structure of the native ventricle. In each well of the platform, an embedded flexible strain gauge provides continuous and non-invasive readout of the contractile stress and beat rate of an engineered cardiac tissue. The sensors are based on micro-cracked titanium-gold thin films, which ensure that the sensors are highly compliant and robust. We demonstrate the value of the platform for toxicology and drug-testing purposes by p...
In vitro disease models offer the ability to study specific systemic features in isolation to bet... more In vitro disease models offer the ability to study specific systemic features in isolation to bet-ter understand underlying mechanisms that lead to dysfunction. Here, we present a cardiac dysfunction model using angiotensin II (ANG II) to elicit pathological responses in a heart-on-a-chip platform that recapitulates native laminar cardiac tissue structure. Our platform, composed of arrays of muscular thin films (MTF), allows for functional comparisons of healthy and diseased tissues by tracking film deflections resulting from contracting tissues. To test our model, we measured gene expression profiles, morphological remodeling, cal-cium transients, and contractile stress generation in response to ANG II exposure and com-pared against previous experimental and clinical results. We found that ANG II induced pathological gene expression profiles including over-expression of natriuretic peptide B, Rho GTPase 1, and T-type calcium channels. ANG II exposure also increased proarrhyth-mic e...
Biomedical research has relied on animal studies and conventional cell cultures for decades. Rece... more Biomedical research has relied on animal studies and conventional cell cultures for decades. Recently, microphysiological systems (MPS), also known as organs-on-chips, that recapitulate the structure and function of native tissues in vitro, have emerged as a promising alternative. However, current MPS typically lack integrated sensors and their fabrication requires multi-step lithographic processes. Here, we introduce a facile route for fabricating anewclass of instrumented cardiacmicrophysiological devicesviamultimaterial three-dimensional (3D) printing. Specifically, we designed six functional inks, based on piezo-resistive, high-conductance, and biocompatible soft materials that enable integration of soft strain gauge sensors within micro-architectures that guide the self-assembly of physio-mimetic laminar cardiac tissues. We validated that these embedded sensors provide non-invasive, electronic readouts of tissue contractile stresses inside cell incubator environments. We furthe...
Extracellular vesicles (EVs) derived from various stem cell sources induce cardioprotective effec... more Extracellular vesicles (EVs) derived from various stem cell sources induce cardioprotective effects during ischemia-reperfusion injury (IRI). These have been attributed mainly to the antiapoptotic, proangiogenic, microRNA (miRNA) cargo within the stem cell–derived EVs. However, the mechanisms of EV-mediated endothelial signaling to cardiomyocytes, as well as their therapeutic potential toward ischemic myocardial injury, are not clear. EV content beyond miRNA that may contribute to cardioprotection has not been fully illuminated. This study characterized the protein cargo of human vascular endothelial EVs (EEVs) to identify lead cardioactive proteins and assessed the effect of EEVs on human laminar cardiac tissues (hlCTs) exposed to IRI. We mapped the protein content of human vascular EEVs and identified proteins that were previously associated with cellular metabolism, redox state, and calcium handling, among other processes. Analysis of the protein landscape of human cardiomyocytes...
Biomedical research has relied on animal studies and conventional cell cultures for decades. Rece... more Biomedical research has relied on animal studies and conventional cell cultures for decades. Recently, microphysiological systems (MPS), also known as organs-on-chips, that recapitulate the structure and function of native tissues in vitro, have emerged as a promising alternative. However, current MPS typically lack integrated sensors and their fabrication requires multi-step lithographic processes. Here, we introduce a facile route for fabricating a new class of instrumented cardiac microphysiological devices via multimaterial three-dimensional (3D) printing. Specifically, we designed six functional inks, based on piezo-resistive, high-conductance, and biocompatible soft materials that enable integration of soft strain gauge sensors within micro-architectures that guide the self-assembly of physio-mimetic laminar cardiac tissues. We validated that these embedded sensors provide non-invasive, electronic readouts of tissue contractile stresses inside cell incubator environments. We f...
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Papers by moran yadid