Heart on a chip: Micro-nanofabrication and microfluidics steering the future of cardiac tissue engineering

The evolution of micro and nanofabrication approaches significantly spurred the advancements of cardiac tissue engineering over the last decades. Engineering in the micro and nanoscale allows for the rebuilding of heart tissues using cardiomyocytes. The breakthrough of human induced pluripotent stem cells expanded this field rendering the development of human tissues from adult cells possible, thus avoiding the ethical issues of the usage of embryonic stem cells but also creating patient-specific human engineered tissues. In the case of the heart, the combination of cardiomyocytes derived from human induced pluripotent stem cells and micro/nano engineering devices gave rise to new therapeutic approaches of cardiac diseases. In this review, we survey the micro and nanofabrication methods used for cardiac tissue engineering, ranging from clean room-based patterning (such as photolithography and plasma etching) to electrospinning and additive manufacturing. Subsequently, we report on the main approaches of microfluidics for cardiac culture systems, the so-called “Heart on a Chip”, and we assess their efficacy for future development of cardiac disease modeling and drug screening platforms

Coating particles using liquids and foams based on viscous formulations with industrial mixers: Batch operation

Particle or powder coating with viscous liquids has been essential in industry for surface modification purposes to induce and enhance specific functionalities. This paper evaluates the performance of using foams (of different bubble diameters) versus liquids as a means of coating powder beds based on viscous liquid formulations. Coating with viscous liquids present numerous industrial challenges and therefore preparing foam equivalents can render the liquid component weak enough (through pre shearing to form the foam), to allow it to break up and coat particles under the shear forces exerted in a mixing device. In this study, two shear mixers are used; the first type consists of paddles in different configurations attached to a single rotating shaft, whilst the second type is a commercial twin screw mixer (TSM). The quality of coating achieved on the bulk powder bed using liquids and foams (stained with a dye) is assessed by image analysis to determine the homogeneity of the color distribution. In addition, scanning electron microscopy provides a tool to further investigate the coating quality of individual particles from the bulk product. The results show that large bubble (centimeter size) foams are much more effective at distributing within the fluidized powder bed compared to the starting viscous liquid and small bubble foams (sub-millimeter size). Furthermore, there is a maximum ratio of foam to powder beyond which agglomeration occurs and is insufficient to fully coat the particles. Coating of individual particles is achieved in the case of the TSM, whereas SEM proves that the single shaft paddle mixer crushes the particles and subsequently granulates them together to form granules of a size comparable to the size of coated particles seen after coating with the TSM

The Quest for the Holy Grail Of 3D Printing: A Critical Review of Recycling in Polymer Powder Bed Fusion Additive Manufacturing

The benefits of additive manufacturing (AM) are widely recognised, boosting the AM method’s use in industry, while it is predicted AM will dominate the global manufacturing industry. Alas, 3D printing’s growth is hindered by its sustainability. AM methods generate vast amounts of residuals considered as waste, which are disposed of. Additionally, the energy consumed, the materials used, and numerous other factors render AM unsustainable. This paper aims to bring forward all documented solutions in the literature. The spotlight is on potential solutions for the Powder Bed Fusion (PBF) AM, focusing on Selective Laser Sintering (SLS), as these are candidates for mass manufacturing by industry. Solutions are evaluated critically, to identify research gaps regarding the recyclability of residual material. Only then can AM dominate the manufacturing industry, which is extremely important since this is a milestone for our transition into sustainable manufacturing. This transition itself is a complex bottleneck on our quest for becoming a sustainable civilisation. Unlike previous reviews that primarily concentrate on specific AM recycling materials, this paper explores the state of the art in AM recycling processes, incorporating the latest market data and projections. By offering a holistic and forward-looking perspective on the evolution and potential of AM, this review serves as a valuable resource for researchers and industry professionals alike

Aging Dynamics in Polymer Powder Bed Fusion Systems: The Case of Selective Laser Sintering

Additive manufacturing (AM) is an advanced production method for layer-by-layer fabrication, offering a paradigm shift in manufacturing. However, the sustainability of AM processes is poor, since suppliers recommend reusing 50 %-70 % of reprocessed powder, contributing to a significant increase in material disposal. To explore the possibility of fully reusing the polymeric material, we conduct a comprehensive characterisation of the powder particulates, in combination with analysis of the final prints. Utilizing optical and scanning electron microscopes, we statistically evaluate the size, morphology, and shape of the particles. Furthermore, tensile strength and deformation of printed bars is evaluated, showcasing the impact of aging on the print properties. The findings reveal that consecutive reuse of used powder significantly influences dimensional accuracy of the printed parts. We detect a 30.63 % relative value of shrinkage after six printing iterations, which corresponds to an absolute shrinkage increase by 0.98 %. This is significant considering the standard shrinkage for the material used is already 3.2 %. Additionally, parts that are printed with reused material exhibit a small increase in elongation at yield, as well as an unexpected rise in tensile strength. Significant agglomeration of small particles is observed in the aged powder, since there are particles of less than 10 µm, which are not found in the virgin powder. These results contribute to a better understanding of the issues related to the reusing of aged material, and offer invaluable insights for mitigating the environmental impact that is associated with material disposal in AM

Sample deposition onto cryo-EM grids: from sprays to jets and back

Despite the great strides made in the field of single-particle cryogenic electron microscopy (cryo-EM) in microscope design, direct electron detectors and new processing suites, the area of sample preparation is still far from ideal. Traditionally, sample preparation involves blotting, which has been used to achieve high resolution, particularly for well behaved samples such as apoferritin. However, this approach is flawed since the blotting process can have adverse effects on some proteins and protein complexes, and the long blot time increases exposure to the damaging air-water interface. To overcome these problems, new blotless approaches have been designed for the direct deposition of the sample on the grid. Here, different methods of producing droplets for sample deposition are compared. Using gas dynamic virtual nozzles, small and high-velocity droplets were deposited on cryo-EM grids, which spread sufficiently for high-resolution cryo-EM imaging. For those wishing to pursue a similar approach, an overview is given of the current use of spray technology for cryo-EM grid preparation and areas for enhancement are pointed out. It is further shown how the broad aspects of sprayer design and operation conditions can be utilized to improve grid quality reproducibly

Coating powder beds with liquids and foams based on viscous formulations using a twin screw mixer: A continuous process study

Coating with viscous formulations has been essential in numerous industries as it can be a means for providing functionalization, additional properties, as well as other benefits. However, there have been scarce studies that have investigated and proposed methodologies in literature. Continuous coating of powders with viscous liquids poses as a promising technology, which has been mentioned in some studies, but has not yet been thoroughly investigated. This paper employs the use of image processing and analysis, in combination with statistical analysis of particles to evaluate the effectiveness of foams and liquids as a means of coating powder beds. Two different sizes of twin screw mixers that are working in continuous operation are employed, and a new continuous foaming device is fabricated and used for the experiments of coating. The effect of materials and process parameters (as for example rotational speed, and flowrate) on the quality of coating are investigated. Image analysis is used to assess the coating quality. The results clearly showcase the potential of using twin screw mixers for coating purposes and not only for mixing. The hypothesis that using large bubble foams to improve the coating of viscous liquids on particles is proven correct, as they provide higher quality coatings compared to their equivalent liquids, when used in the twin screw mixer. Surprisingly, using a larger scale twin screw mixer, does not show a substantial effect on the mixing, regarding quality, however there is still a requirement for mix optimization for achieving scale-up of this process. These results provide a new pathway for coating powders with viscous formulations in industrial applications, requiring less energy and effort in this process, and can pave the way towards introducing more sustainable industrial methodologies for coating

A cryo-EM grid preparation device for time-resolved structural studies

Structural biology generally provides static snapshots of protein conformations that can provide information on the functional mechanisms of biological systems. Time-resolved structural biology provides a means to visualize, at near-atomic resolution, the dynamic conformational changes that macromolecules undergo as they function. X-ray free-electron-laser technology has provided a powerful tool to study enzyme mechanisms at atomic resolution, typically in the femtosecond to picosecond timeframe. Complementary to this, recent advances in the resolution obtainable by electron microscopy and the broad range of samples that can be studied make it ideally suited to time-resolved approaches in the microsecond to millisecond timeframe to study large loop and domain motions in biomolecules. Here we describe a cryo-EM grid preparation device that permits rapid mixing, voltage-assisted spraying and vitrification of samples. It is shown that the device produces grids of sufficient ice quality to enable data collection from single grids that results in a sub-4 Å reconstruction. Rapid mixing can be achieved by blot-and-spray or mix-and-spray approaches with a delay of ∼10 ms, providing greater temporal resolution than previously reported mix-and-spray approaches