In tissues and organs, the extracellular matrix (ECM) helps maintain inter- and intracellular architectures that sustain the structure–function relationships defining physiological homeostasis. Combining fiber scaffolds and cells to form engineered tissues is a means of replicating these relationships. Engineered tissues' fiber scaffolds are designed to mimic the topology and chemical composition of the ECM network. Here, we asked how cells found in the heart compare in their propensity to align their cytoskeleton and self-organize in response to topological cues in fibrous scaffolds. We studied cardiomyocytes, valvular interstitial cells, and vascular endothelial cells as they adapted their inter- and intracellular architectures to the extracellular space. We used focused rotary jet spinning to manufacture aligned fibrous scaffolds to mimic the length scale and three-dimensional (3D) nature of the native ECM in the muscular, valvular, and vascular tissues of the heart. The representative cardiovascular cell types were seeded onto fiber scaffolds and infiltrated the fibrous network. We measured different cell types' propensity for cytoskeletal alignment in response to fiber scaffolds with differing levels of anisotropy. The results indicated that valvular interstitial cells on moderately anisotropic substrates have a higher propensity for cytoskeletal alignment than cardiomyocytes and vascular endothelial cells. However, all cell types displayed similar levels of alignment on more extreme (isotropic and highly anisotropic) fiber scaffold organizations. These data suggest that in the hierarchy of signals that dictate the spatiotemporal organization of a tissue, geometric cues within the ECM and cellular networks may homogenize behaviors across cell populations and demographics.

Combat helmets are designed to protect against ballistic threats and fragments of explosive devices. There are numerous types of helmet comfort foams available. However, pad systems have not been evaluated in combat helmets to understand to what extent they mitigate head accelerations. In this work, different pad systems are studied to analyze the ballistic performance of combat helmets using a Hybrid III dummy equipped with longitudinal accelerometers at the head and a neck simulator. The tests are conducted with 9 mm Full Metal Jacket (FMJ) projectiles according to the performance requirements III-A of the NIJ 0106.01 standard. This experimental methodology allows the evaluation of brain and neck injuries. The thicker bicomponent polyurethane foams and the honeycomb configuration provided the best results in terms of mitigating brain damage due to accelerations applying different criteria (PLA, WSTC, HIC). However, it was concluded that there is no cervical injury or cranial fracture risk for any of the cases studied.

Combat helmets are the primary system for protecting the head against ballistic impacts. Generally, combat helmets have been evaluated using a ballistic plasticine head surrogate based on international standards. More realistic human head models have recently been introduced to assess combat helmet performance considering biomechanical requirements. In this work, the Hybrid III dummy head and neck has been introduced to evaluate the performance of the combat helmet against the ballistic impact of live ammunition at different impact locations, considering two different thicknesses of the padding system. A numerical model including a helmet and a Hybrid III head and neck, is developed and validated with our experimental data. The results reveal the influence of the location, where the rear impact leads to the highest risk of brain damage. The effect of pad thickness is closely related to the energy absorbed by the helmet, the backface deformation (BFD), the contact force and the acceleration measured on the head.

Arrhythmogenic cardiomyopathy (ACM) is an inherited cardiac disorder that causes life-threatening arrhythmias and myocardial dysfunction. Pathogenic variants in Plakophilin-2 (PKP2), a desmosome component within specialized cardiac cell junctions, cause the majority of ACM cases. However, the molecular mechanisms by which PKP2 variants induce disease phenotypes remain unclear. Here we built bioengineered platforms using genetically modified human induced pluripotent stem cell-derived cardiomyocytes to model the early spatiotemporal process of cardiomyocyte junction assembly in vitro. Heterozygosity for truncating variant PKP2^R413X reduced Wnt/β-catenin signaling, impaired myofibrillogenesis, delayed mechanical coupling, and reduced calcium wave velocity in engineered tissues. These abnormalities were ameliorated by SB216763, which activated Wnt/β-catenin signaling, improved cytoskeletal organization, restored cell junction integrity in cell pairs, and improved calcium wave velocity in engineered tissues. Together, these findings highlight the therapeutic potential of modulating Wnt/β-catenin signaling in a human model of ACM.

Hydrogels are attractive materials for tissue engineering, but efforts to date have shown limited ability to produce the microstructural features necessary to promote cellular self-organization into hierarchical three-dimensional (3D) organ models. Here we develop a hydrogel ink containing prefabricated gelatin fibres to print 3D organ-level scaffolds that recapitulate the intra- and intercellular organization of the heart. The addition of prefabricated gelatin fibres to hydrogels enables the tailoring of the ink rheology, allowing for a controlled sol–gel transition to achieve precise printing of free-standing 3D structures without additional supporting materials. Shear-induced alignment of fibres during ink extrusion provides microscale geometric cues that promote the self-organization of cultured human cardiomyocytes into anisotropic muscular tissues in vitro. The resulting 3D-printed ventricle in vitro model exhibited biomimetic anisotropic electrophysiological and contractile properties.

Children worldwide suffer from heart valve disease and often require open heart surgeries for valve replacements. Unfortunately, current heart valve replacements do not grow alongside the child, necessitating repeat high-risk surgeries throughout the pediatric patient’s life. This work introduces FibraValves, heart valve replacements fabricated in minutes that comprise of fibers produced by focused rotary jet spinning. FibraValves are manufactured using biodegradable polymer fibers that allow for the patient’s cells to attach and remodel the implanted scaffold, eventually building a native valve that can grow and live with the child throughout their life. These valves were tested in vitro and deployed in acute in vivo studies to evaluate their ability to maintain unidirectional blood flow in the heart. Together, these results suggest the potential translation of FibraValves as future cardiac implants, eliminating the need for repeated valve replacements in children.

Light is recognized as an accurate and noninvasive tool for stimulating excitable cells. Here, we report on a non-genetic approach based on
organic molecular phototransducers that allows wiring- and electrode-free tissue modulation. As a proof of concept, we show photostimulation
of an in vitro cardiac microphysiological model mediated by an amphiphilic azobenzene compound that preferentially dwells in the cell membrane.
Exploiting this optical based stimulation technology could be a disruptive approach for highly resolved cardiac tissue stimulation.

Helical alignments within the heart’s musculature have been speculated to be important in achieving physiological pumping efficiencies. Testing this possibility is difficult, however, because it is challenging to reproduce the fine spatial features and complex structures of the heart’s musculature using current techniques. Here we report focused rotary jet spinning (FRJS), an additive manufacturing approach that enables rapid fabrication of micro/nanofiber scaffolds with programmable alignments in three-dimensional geometries. Seeding these scaffolds with cardiomyocytes enabled the biofabrication of tissue-engineered ventricles, with helically aligned models displaying more uniform deformations, greater apical shortening, and increased ejection fractions compared with circumferential alignments. The ability of FRJS to control fiber arrangements in three dimensions offers a streamlined approach to fabricating tissues and organs, with this work demonstrating how helical architectures contribute to cardiac performance.

Food waste and food safety motivate the need for improved food packaging solutions. However, current films/coatings addressing these issues are often limited by inefficient release dynamics that require large quantities of active ingredients. Here we developed antimicrobial pullulan fibre (APF)-based packaging that is biodegradable and capable of wrapping food substrates, increasing their longevity and enhancing their safety. APFs were spun using a high-throughput system, termed focused rotary jet spinning, with water as the only solvent, allowing the incorporation of naturally derived antimicrobial agents. Using avocados as a representative example, we demonstrate that APF-coated samples had their shelf life extended by inhibited proliferation of natural microflora, and lost less weight than uncoated control samples. This work offers a promising technique to produce scalable, low-cost and environmentally friendly biodegradable antimicrobial packaging systems.

An ensemble of in vitro cardiac tissue models has been developed over the past several decades to aid our understanding of complex cardiovascular disorders using a reductionist approach. These approaches often rely on recapitulating single or multiple clinically relevant end points in a dish indicative of the cardiac pathophysiology. The possibility to generate disease-relevant and patient-specific human induced pluripotent stem cells has further leveraged the utility of the cardiac models as screening tools at a large scale. To elucidate biological mechanisms in the cardiac models, it is critical to integrate physiological cues in form of biochemical, biophysical, and electromechanical stimuli to achieve desired tissue-like maturity for a robust phenotyping. Here, we review the latest advances in the directed stem cell differentiation approaches to derive a wide gamut of cardiovascular cell types, to allow customization in cardiac model systems, and to study diseased states in multiple cell types. We also highlight the recent progress in the development of several cardiovascular models, such as cardiac organoids, microtissues, engineered heart tissues, and microphysiological systems. We further expand our discussion on defining the context of use for the selection of currently available cardiac tissue models. Last, we discuss the limitations and challenges with the current state-of-the-art cardiac models and highlight future directions.

Engineered nanomaterials offer the benefit of having systematically tunable physicochemical characteristics (e.g., size, dimensionality, and surface chemistry) that highly dictate the biological activity of a material. Among the most promising engineered nanomaterials to date are graphene-family nanomaterials (GFNs), which are 2-D nanomaterials (2DNMs) with unique electrical and mechanical properties. Beyond engineering new nanomaterial properties, employing safety-by-design through considering the consequences of cell-material interactions is essential for exploring their applicability in the biomedical realm. In this study, we asked the effect of GFNs on the endothelial barrier function and cellular architecture of vascular endothelial cells. Using micropatterned cell pairs as a reductionist in vitro model of the endothelium, the progression of cytoskeletal reorganization as a function of GFN surface chemistry and time was quantitatively monitored. Here, we show that the surface oxidation of GFNs (graphene, reduced graphene oxide, partially reduced graphene oxide, and graphene oxide) differentially affect the endothelial barrier at multiple scales; from the biochemical pathways that influence the development of cellular protrusions to endothelial barrier integrity. More oxidized GFNs induce higher endothelial permeability and the increased formation of cytoplasmic protrusions such as filopodia. We found that these changes in cytoskeletal organization, along with barrier function, can be potentiated by the effect of GFNs on the Rho/Rho-associated kinase (ROCK) pathway. Specifically, GFNs with higher surface oxidation elicit stronger ROCK2 inhibitory behavior as compared to pristine graphene sheets. Overall, findings from these studies offer a new perspective towards systematically controlling the surface-dependent effects of GFNs on cytoskeletal organization via ROCK2 inhibition, providing insight for implementing safety-by-design principles in GFN manufacturing towards their targeted biomedical applications.

Biohybrid systems have been developed to better understand the design principles and coordination mechanisms of biological systems. We consider whether two functional regulatory features of the heart—mechanoelectrical signaling and automaticity—could be transferred to a synthetic analog of another fluid transport system: a swimming fish. By leveraging cardiac mechanoelectrical signaling, we recreated reciprocal contraction and relaxation in a muscular bilayer construct where each contraction occurs automatically as a response to the stretching of an antagonistic muscle pair. Further, to entrain this closed-loop actuation cycle, we engineered an electrically autonomous pacing node, which enhanced spontaneous contraction. The biohybrid fish equipped with intrinsic control strategies demonstrated self-sustained body–caudal fin swimming, highlighting the role of feedback mechanisms in muscular pumps such as the heart and muscles.

The unfolded states of fibronectin (FN) subsequently induce the formation of an extracellular matrix (ECM) fibrillar network, which is necessary to generate new substitutive tissues. Here, the authors demonstrate that negatively charged small unilamellar vesicles (SUVs) qualify as candidates for FN delivery due to their remarkable effects on the autonomous binding and unfolding of FN, which leads to increased tissue regeneration. In vitro experiments revealed that the FN-SUV complex remarkably increased the attachment, differentiation, and migration of fibroblasts. The potential utilization of this complex in vivo to treat inflammatory colon diseases is also described based on results obtained for ameliorated conditions in rats with ulcerative colitis (UC) that had been treated with the FN-SUV complex. Their findings provide a new ECM-delivery platform for ECM-based therapeutic applications and suggest that properly designed SUVs may be an unprecedented FN-delivery system that is highly effective in treating UC and inflammatory bowel diseases.

Duchenne muscular dystrophy (DMD) is a devastating genetic disease leading to degeneration of skeletal muscles and premature death. How dystrophin absence leads to muscle wasting remains unclear. Here, we describe an optimized protocol to differentiate human induced pluripotent stem cells (iPSC) to a late myogenic stage. This allows us to recapitulate classical DMD phenotypes (mislocalization of proteins of the dystrophin-associated glycoprotein complex, increased fusion, myofiber branching, force contraction defects, and calcium hyperactivation) in isogenic DMD-mutant iPSC lines in vitro. Treatment of the myogenic cultures with prednisolone (the standard of care for DMD) can dramatically rescue force contraction, fusion, and branching defects in DMD iPSC lines. This argues that prednisolone acts directly on myofibers, challenging the largely prevalent view that its beneficial effects are caused by antiinflammatory properties. Our work introduces a human in vitro model to study the onset of DMD pathology and test novel therapeutic approaches.

Diabetes is a disease of insulin insufficiency, requiring many to rely on exogenous insulin with constant monitoring to avoid a fatal outcome. Islet transplantation is a recent therapy that can provide insulin independence, but the procedure is still limited by both the availability of human islets and reliable tests to assess their function. While stem cell technologies are poised to fill the shortage of transplantable cells, better methods are still needed for predicting transplantation outcome. To ensure islet quality, we propose that the next generation of islet potency tests should be biomimetic systems that match glucose stimulation dynamics and cell microenvironmental preferences and rapidly assess conditional and continuous insulin secretion with minimal manual handing. Here, we review the current approaches for islet potency testing and outline technologies and methods that can be used to arrive at a more predictive potency test that tracks islet secretory capacity in a relevant context. With the development of potency tests that can report on islet secretion dynamics in a context relevant to their intended function, islet transplantation can expand into a more widely accessible and reliable treatment option for individuals with diabetes.

High-voltage electrical fields and low production rate limit electrospinning, the electrical charging of polymer liquids, as a means of nanofiber fabrication. Here, we show a facile method of fabrication of aligned three-dimensional nanofiber structures by utilizing high-speed, rotating polymer solution jets to extrude fibers. Termed rotary jet-spinning, fiber morphology, diameter, and web porosity can be controlled by varying nozzle geometry, rotation speed, and polymer solution properties. We demonstrate the utility of this technique for tissue engineering by building anisotropic arrays of biodegradable polymer fibers and seeding the constructs with neonatal rat ventricular cardiomyocytes. The myocytes used the aligned fibers to orient their contractile cytoskeleton and to self-organize into a beating, multicellular tissue that mimics the laminar, anisotropic architecture of the heart muscle. This technique may prove advantageous for building uniaxially aligned nanofiber structures for polymers which are not amenable to fabrication by electrospinning.

My second tour of duty in the Global War on Terror took me, an Army reservist, from my research and teaching at Harvard to the mountainous valleys of eastern Afghanistan. My job on this tour, working for the Center for Army Lessons Learned, was to try to understand how we were trying to rid the battlefield of improvised explosive devices, or IEDs, which were causing most of the Coalition casualties. A field-grade officer with no troops under my command, I roamed my area of operation attached to tactical units to collect data. One such group was Route Clearance Patrol 13 (RCP13), an amalgam of combat engineers, explosive specialists, and specialized teams, clearing roads of IEDs (Figure 1). On these missions, I sat strapped into its Buffalo, a motorized beast whose extraordinary size and robotic arm capabilities were designed to directly engage the explosive devices in lieu of dispatching someone to check it out themselves.

It is our pleasure to welcome you to the first issue of Biophysics Reviews (BPR), a new journal from AIP Publishing covering the diverse field of biophysics. The journal expands on the tradition of excellence set by Applied Physics Reviews (APR) by publishing high impact, cutting edge research and reviews that are valuable for both emerging and experienced researchers.

Extracellular vesicles, small membrane-bound particles released from cells, have been shown to have cardioprotective effects. Here, Yadid et al. analyzed the proteins contained in vesicles released from endothelial cells under normoxia and hypoxia and investigated cardioprotective effects on cardiac tissues in vitro. Using a human heart-on-chip composed of cardiomyocytes, the authors showed that endothelial cell–derived vesicles supported metabolic function, tissue contraction, and viability during ischemia-reperfusion injury. This study helps to elucidate the mechanism by which vesicles are cardioprotective in human tissue.

There is an urgent need to develop biodegradable and nontoxic materials from biopolymers and nature-derived antimicrobials to enhance food safety and quality. In this study, electrospinning was used as a one-step, scalable, green synthesis approach to engineer antimicrobial fibers from zein using nontoxic organic solvents and a cocktail of nature-derived antimicrobials which are all FDA-classified Generally Recognized as Safe (GRAS) for food use. Morphological and physicochemical properties of fibers, as well as the dissolution kinetics of antimicrobials were assessed along with their antimicrobial efficacy using state of the art analytical and microbiological methods. A cocktail of nature-derived antimicrobials was developed and included thyme oil, citric acid, and nisin. Its ability to inactivate a broad-spectrum of with food-related pathogens was demonstrated. Morphological characterization of the electrospun antimicrobial fibers revealed bead-free fibers with a small average diameter of 165 nm, whereas physicochemical characterization showed high surface area-to-volume ratio (specific surface area:21.91 m2/g) and presence of antimicrobial analytes in the fibers. The antimicrobials exhibited initial rapid release from the fibers in 2 h into various food simulants. Furthermore, the antimicrobial fibers effectively reduced E. coli and L. innocua populations by ∼5 logs for after 24 h and 1 h of exposure, respectively. More importantly, due to the small diameter and high surface area-to-volume ratio of the fibers, only miniscule quantities of fiber mass and antimicrobials per surface area (2.50 mg/cm2 of fibers) are needed for pathogen inactivation. The scalability of this fiber synthesis process was also demonstrated using a multineedle injector with production yield up to 1 g/h. This study shows the potential of using nature-derived biopolymers and antimicrobials to synthesize fibers for sustainable food packaging materials.