Low-cost, versatile, and highly reproducible microfabrication pipeline to generate 3D-printed customised cell culture devices with complex designs.

Cell culture devices, such as microwells and microfluidic chips, are designed to increase the complexity of cell-based models while retaining control over culture conditions and have become indispensable platforms for biological systems modelling. From microtopography, microwells, plating devices, and microfluidic systems to larger constructs such as live imaging chamber slides, a wide variety of culture devices with different geometries have become indispensable in biology laboratories. However, while their application in biological projects is increasing exponentially, due to a combination of the techniques, equipment and tools required for their manufacture, and the expertise necessary, biological and biomedical labs tend more often to rely on already made devices. Indeed, commercially developed devices are available for a variety of applications but are often costly and, importantly, lack the potential for customisation by each individual lab. The last point is quite crucial, as often experiments in wet labs are adapted to whichever design is already available rather than designing and fabricating custom systems that perfectly fit the biological question. This combination of factors still restricts widespread application of microfabricated custom devices in most biological wet labs. Capitalising on recent advances in bioengineering and microfabrication aimed at solving these issues, and taking advantage of low-cost, high-resolution desktop resin 3D printers combined with PDMS soft lithography, we have developed an optimised a low-cost and highly reproducible microfabrication pipeline. This is thought specifically for biomedical and biological wet labs with not prior experience in the field, which will enable them to generate a wide variety of customisable devices for cell culture and tissue engineering in an easy, fast reproducible way for a fraction of the cost of conventional microfabrication or commercial alternatives. This protocol is designed specifically to be a resource for biological labs with limited expertise in those techniques and enables the manufacture of complex devices across the µm to cm scale. We provide a ready-to-go pipeline for the efficient treatment of resin-based 3D-printed constructs for PDMS curing, using a combination of polymerisation steps, washes, and surface treatments. Together with the extensive characterisation of the fabrication pipeline, we show the utilisation of this system to a variety of applications and use cases relevant to biological experiments, ranging from micro topographies for cell alignments to complex multipart hydrogel culturing systems. This methodology can be easily adopted by any wet lab, irrespective of prior expertise or resource availability and will enable the wide adoption of tailored microfabricated devices across many fields of biology.

A 3D-Printed Hybrid Nasal Cartilage with Functional Electronic Olfaction.

Advances in biomanufacturing techniques have opened the doors to recapitulate human sensory organs such as the nose and ear in vitro with adequate levels of functionality. Such advancements have enabled simultaneous targeting of two challenges in engineered sensory organs, especially the nose: i) mechanically robust reconstruction of the nasal cartilage with high precision and ii) replication of the nose functionality: odor perception. Hybrid nasal organs can be equipped with remarkable capabilities such as augmented olfactory perception. Herein, a proof-of-concept for an odor-perceptive nose-like hybrid, which is composed of a mechanically robust cartilage-like construct and a biocompatible biosensing platform, is proposed. Specifically, 3D cartilage-like tissue constructs are created by multi-material 3D bioprinting using mechanically tunable chondrocyte-laden bioinks. In addition, by optimizing the composition of stiff and soft bioinks in macro-scale printed constructs, the competence of this system in providing improved viability and recapitulation of chondrocyte cell behavior in mechanically robust 3D constructs is demonstrated. Furthermore, the engineered cartilage-like tissue construct is integrated with an electrochemical biosensing system to bring functional olfactory sensations toward multiple specific airway disease biomarkers, explosives, and toxins under biocompatible conditions. Proposed hybrid constructs can lay the groundwork for functional bionic interfaces and humanoid cyborgs.

Bacterial Nanobionics via 3D Printing.

Investigating the multidimensional integration between different microbiological kingdoms possesses potential toward engineering next-generation bionic architectures. Bacterial and fungal kingdom exhibits mutual symbiosis that can offer advanced functionalities to these bionic architectures. Moreover, functional nanomaterials can serve as probing agents for accessing newer information from microbial organisms due to their dimensional similarities. In this article, a bionic mushroom was created by intertwining cyanobacterial cells with graphene nanoribbons (GNRs) onto the umbrella-shaped pileus of mushroom for photosynthetic bioelectricity generation. These seamlessly merged GNRs function as agents for mediating extracellular electron transport from cyanobacteria resulting in photocurrent generation. Additionally, three-dimensional (3D) printing technique was used to assemble cyanobacterial cells in anisotropic, densely packed geometry resulting in adequate cell-population density for efficient collective behavior. These 3D printed cyanobacterial colonies resulted in comparatively higher photocurrent (almost 8-fold increase) than isotropically casted cyanobacteria of similar seeding density. An insight of the proposed integration between cyanobacteria and mushroom derives remarkable advantage that arises from symbiotic relationship, termed here as engineered bionic symbiosis. Existence of this engineered bionic symbiosis was confirmed by UV-visible spectroscopy and standard plate counting method. Taken together, the present study augments scientific understanding of multidimensional integration between the living biological microworld and functional abiotic nanomaterials to establish newer dimensionalities toward advancement of bacterial nanobionics.

Tailoring Thin-Film Piezoelectrics for Crash Sensing.

Crash sensing and its assessment play a pivotal role in autonomous vehicles for preventing fatal casualties. Existing crash sensors are severely bottlenecked by sluggish response time, rigid mechanical components, and space constraints. Miniaturized sensors embedded with custom-tailored nanomaterials upholds potential to overcome these limitations. In this article, piezoelectric Zinc-Oxide thin film as a crash sensing layer is integrated onto a flexible metal-alloy cantilever. Material characterization studies are conducted to confirm piezoelectric property of sputtered ZnO film. The piezoelectric d 31 coefficient value of ZnO film was 7.2 pm V-1 . The ZnO sensing element is firmly mounted on a scaled car model and used in a crash sensing experimental set-up. A comprehensive theoretical analysis for two different real scenarios (nearly elastic and nearly inelastic collision) of crash events followed by experimental study is discussed. The crash sensor's output exhibits a linear relationship with magnitude of impact forces experienced at crash events. The response time of ZnO crash sensor is 18.2 ms, and it exhibits a sensitivity of 28.7 mV N-1 . The developed crash sensor has potential to replace bulk material sensors owing to its faster response time, high sensitivity, and compactness as the demand for crash sensors in next-generation automobile industries is progressively growing.

Evaluation of transverse piezoelectric coefficient of ZnO thin films deposited on different flexible substrates: a comparative study on the vibration sensing performance.

We report on the systematic comparative study of highly c-axis oriented and crystalline piezoelectric ZnO thin films deposited on four different flexible substrates for vibration sensing application. The flexible substrates employed for present experimental study were namely a metal alloy (Phynox), metal (aluminum), polyimide (Kapton), and polyester (Mylar). ZnO thin films were deposited by an RF reactive magnetron sputtering technique. ZnO thin films of similar thicknesses of 700 ± 30 nm were deposited on four different flexible substrates to have proper comparative studies. The crystallinity, surface morphology, chemical composition, and roughness of ZnO thin films were evaluated by respective material characterization techniques. The transverse piezoelectric coefficient (d31) value for assessing the piezoelectric property of ZnO thin films on different flexible substrates was measured by a four-point bending method. ZnO thin films deposited on Phynox alloy substrate showed relatively better material characterization results and a higher piezoelectric d31 coefficient value as compared to ZnO films on metal and polymer substrates. In order to experimentally verify the above observations, vibration sensing studies were performed. As expected, the ZnO thin film deposited on Phynox alloy substrate showed better vibration sensing performance. It has generated the highest peak to peak output voltage amplitude of 256 mV as compared to that of aluminum (224 mV), Kapton (144 mV), and Mylar (46 mV). Therefore, metal alloy flexible substrate proves to be a more suitable, advantageous, and versatile choice for integrating ZnO thin films as compared to metal and polymer flexible substrates for vibration sensing applications. The present experimental study is extremely important and helpful for the selection of a suitable flexible substrate for various applications in the field of sensor and actuator technology.