John F. Zimmerman

Free-Standing Kinked Silicon Nanowires for Probing Inter- and Intracellular Force Dynamics

Silicon nanowires (SiNWs) have emerged as a new class of materials with important applications in biology and medicine with current efforts having focused primarily on using substrate bound SiNW devices. However, developing devices capable of free-standing inter- and intracellular operation is an important next step in designing new synthetic cellular materials and tools for biophysical characterization. To demonstrate this, here we show that label free SiNWs can be internalized in multiple cell lines, forming robust cytoskeletal interfaces, and when kinked can serve as free-standing inter- and intracellular force probes capable of continuous extended (>1 h) force monitoring. Our results show that intercellular interactions exhibit ratcheting like behavior with force peaks of ∼69.6 pN/SiNW, while intracellular force peaks of ∼116.9 pN/SiNW were recorded during smooth muscle contraction. To accomplish this, we have introduced a simple single-capture dark-field/phase contrast optical imaging modality, scatter enhanced phase contrast (SEPC), which enables the simultaneous visualization of both cellular components and inorganic nanostructures. This approach demonstrates that rationally designed devices capable of substrate-independent operation are achievable, providing a simple and scalable method for continuous inter- and intracellular force dynamics studies.

Cellular uptake and dynamics of unlabeled freestanding silicon nanowires

Cells naturally “eat” nanowires, paving way for intracellular sensing devices and photoresponsive therapies. The ability to seamlessly merge electronic devices with biological systems at the cellular length scale is an exciting prospect for exploring new fundamental cell biology and in designing next-generation therapeutic devices. Semiconductor nanowires are well suited for achieving this goal because of their intrinsic size and wide range of possible configurations. However, current studies have focused primarily on delivering substrate-bound nanowire devices through mechanical abrasion or electroporation, with these bulkier substrates negating many of the inherent benefits of using nanoscale materials. To improve on this, an important next step is learning how to distribute these devices in a drug-like fashion, where cells can naturally uptake and incorporate these electronic components, allowing for truly noninvasive device integration. We show that silicon nanowires (SiNWs) can potentially be used as such a system, demonstrating that label-free SiNWs can be internalized in multiple cell lines (96% uptake rate), undergoing an active “burst-like” transport process. Our results show that, rather than through exogenous manipulation, SiNWs are internalized primarily through an endogenous phagocytosis pathway, allowing cellular integration of these materials. To study this behavior, we have developed a robust set of methodologies for quantitatively examining high–aspect ratio nanowire-cell interactions in a time-dependent manner on both single-cell and ensemble levels. This approach represents one of the first dynamic studies of semiconductor nanowire internalization and offers valuable insight into designing devices for biomolecule delivery, intracellular sensing, and photoresponsive therapies.

Nanoscale semiconductor devices as new biomaterials

Research on nanoscale semiconductor devices will elicit a novel understanding of biological systems. First, we discuss why it is necessary to build interfaces between cells and semiconductor nanoelectronics. Second, we describe some recent molecular biophysics studies with nanowire field effect transistor sensors. Third, we present the use of nanowire transistors as electrical recording devices that can be integrated into synthetic tissues and targeted intra- or extracellularly to study single cells. Lastly, we discuss future directions and challenges in further developing this area of research, which will advance biology and medicine.

Photoelectrochemical modulation of neuronal activity with free-standing coaxial silicon nanowires

Optical methods for modulating cellular behaviour are promising for both fundamental and clinical applications. However, most available methods are either mechanically invasive, require genetic manipulation of target cells or cannot provide subcellular specificity. Here, we address all these issues by showing optical neuromodulation with free-standing coaxial p-type/intrinsic/n-type silicon nanowires. We reveal the presence of atomic gold on the nanowire surfaces, likely due to gold diffusion during the material growth. To evaluate how surface gold impacts the photoelectrochemical properties of single nanowires, we used modified quartz pipettes from a patch clamp and recorded sustained cathodic photocurrents from single nanowires. We show that these currents can elicit action potentials in primary rat dorsal root ganglion neurons through a primarily atomic gold-enhanced photoelectrochemical process.The wireless and photoelectrochemical stimulation of primary rat dorsal root ganglion neurons is demonstrated by shining laser light onto coaxially doped silicon nanowires deposited on the neuronal membrane.

A tissue-engineered scale model of the heart ventricle

Laboratory studies of the heart use cell and tissue cultures to dissect heart function yet rely on animal models to measure pressure and volume dynamics. Here, we report tissue-engineered scale models of the human left ventricle, made of nanofibrous scaffolds that promote native-like anisotropic myocardial tissue genesis and chamber-level contractile function. Incorporating neonatal rat ventricular myocytes or cardiomyocytes derived from human induced pluripotent stem cells, the tissue-engineered ventricles have a diastolic chamber volume of ~500 µl (comparable to that of the native rat ventricle and approximately 1/250 the size of the human ventricle), and ejection fractions and contractile work 50–250 times smaller and 104–108 times smaller than the corresponding values for rodent and human ventricles, respectively. We also measured tissue coverage and alignment, calcium-transient propagation and pressure–volume loops in the presence or absence of test compounds. Moreover, we describe an instrumented bioreactor with ventricular-assist capabilities, and provide a proof-of-concept disease model of structural arrhythmia. The model ventricles can be evaluated with the same assays used in animal models and in clinical settings.Scale models of the human left ventricle made of tissue-engineered nanofibrous scaffolds and primary rat cardiomyocytes or human-stem-cell-derived cardiomyocytes enable the study of contractile function and the modelling of structural arrhythmia.

Mussel-inspired 3D fiber scaffolds for heart-on-a-chip toxicity studies of engineered nanomaterials

AbstractDue to the unique physicochemical properties exhibited by materials with nanoscale dimensions, there is currently a continuous increase in the number of engineered nanomaterials (ENMs) used in consumer goods. However, several reports associate ENM exposure to negative health outcomes such as cardiovascular diseases. Therefore, understanding the pathological consequences of ENM exposure represents an important challenge, requiring model systems that can provide mechanistic insights across different levels of ENM-based toxicity. To achieve this, we developed a mussel-inspired 3D microphysiological system (MPS) to measure cardiac contractility in the presence of ENMs. While multiple cardiac MPS have been reported as alternatives to in vivo testing, most systems only partially recapitulate the native extracellular matrix (ECM) structure. Here, we show how adhesive and aligned polydopamine (PDA)/polycaprolactone (PCL) nanofiber can be used to emulate the 3D native ECM environment of the myocardium. Such nanofiber scaffolds can support the formation of anisotropic and contractile muscular tissues. By integrating these fibers in a cardiac MPS, we assessed the effects of TiO2 and Ag nanoparticles on the contractile function of cardiac tissues. We found that these ENMs decrease the contractile function of cardiac tissues through structural damage to tissue architecture. Furthermore, the MPS with embedded sensors herein presents a way to non-invasively monitor the effects of ENM on cardiac tissue contractility at different time points. These results demonstrate the utility of our MPS as an analytical platform for understanding the functional impacts of ENMs while providing a biomimetic microenvironment to in vitro cardiac tissue samples. Graphical AbstractHeart-on-a-chip integrated with mussel-inspired fiber scaffolds for a high-throughput toxicological assessment of engineered nanomaterials

Nongenetic Optical Methods for Measuring and Modulating Neuronal Response

The ability to probe and modulate electrical signals sensitively at cellular length scales is a key challenge in the field of electrophysiology. Electrical signals play integral roles in regulating cellular behavior and in controlling biological function. From cardiac arrhythmias to neurodegenerative disorders, maladaptive phenotypes in electrophysiology can result in serious and potentially deadly medical conditions. Understanding how to monitor and to control these behaviors precisely and noninvasively represents an important step in developing next-generation therapeutic devices. As we develop a deeper understanding of neural network formation, electrophysiology has the potential to offer fundamental insights into the inner working of the brain. In this Perspective, we explore traditional methods for examining neural function, discuss recent genetic advances in electrophysiology, and then focus on the latest innovations in optical sensing and stimulation of action potentials in neurons. We emphasize nongenetic optical methods, as these provide high spatiotemporal resolution and can be achieved with minimal invasiveness.

Chapter 4:Nanowire Biosensors

Nanowire field-effect transistors (NWFETs) represent diverse and powerful nanostructures for achieving nanoscale electronic interfaces with biological systems. NWFETs exhibit exquisite sensitivity in chemical and biological detection and could form strongly coupled interfaces with cellular components. NWFETs also offer the hope of intervening with biological systems on a molecular scale for unprecedented studies of biophysical dynamics, in a nuanced way that has not been feasible up until this point.