Bioengineering, Volume 6, Issue 4 (December 2019) – 29 articles

Tequila agave bagasse (TAB) is the fibrous waste from the Tequila production process. It is generated in large amounts and its disposal is an environmental problem. Its use as a source of fermentable sugars for biotechnological processes is of interest; thus, it was investigated for the production of polyhydroxybutyrate (PHB) by the xylose-assimilating bacteria Burkholderia sacchari. First, it was chemically hydrolyzed, yielding 20.6 g·L⁻¹ of reducing sugars, with xylose and glucose as the main components (7:3 ratio). Next, the effect of hydrolysis by-products on B. sacchari growth was evaluated. Phenolic compounds showed the highest toxicity (> 60% of growth inhibition). Then, detoxification methods (resins, activated charcoal, laccases) were tested to remove the growth inhibitory compounds from the TAB hydrolysate (TABH). The highest removal percentage (92%) was achieved using activated charcoal (50 g·L⁻¹, pH 2, 4 h). Finally, detoxified TABH was used as the carbon source for the production of PHB in a two-step batch culture, reaching a biomass production of 11.3 g·L⁻¹ and a PHB accumulation of 24 g PHB g⁻¹ dry cell (after 122 h of culture). The polymer structure resulted in a homopolymer of 3-hydroxybutyric acid. It is concluded that the TAB could be hydrolyzed and valorized as a carbon source for producing PHB. Full article

Risk assessments (RAs) are frequently conducted to assess the potential effect of process parameters (PPs) on product quality attributes (e.g., a critical quality attribute (CQA)). To evaluate the PPs criticality the risk priority number (RPN) for each PP is often calculated. This number is generated by the multiplication of three factors: severity, occurrence, and detectability. This mathematical operation may result in some potential errors due to the multiplication of ordinal scaled values and the assumption that the factors contribute equally to the PPs criticality. To avoid these misinterpretations and to assess the out of specification (OOS) probability of the drug substance, we present a novel and straightforward mathematical algorithm. This algorithm quantitatively describes the PPs effect on each CQA assessed within the RA. The transcription of severity and occurrence to model effect sizes and parameters distribution are the key elements of the herein developed approach. This approach can be applied to any conventional RA within the biopharmaceutical industry. We demonstrate that severity and occurrence contribute differently to the PP criticality and compare these results with the RPN number. Detectability is used in a final step to precisely sort the contribution of each factor. To illustrate, we show the misinterpretation risk of the PP critically by using the conventional RPN approach. Full article

The realization of biomimetic microenvironments for cell biology applications such as organ-on-chip, in vitro drug screening, and tissue engineering is one of the most fascinating research areas in the field of bioengineering. The continuous evolution of additive manufacturing techniques provides the tools to engineer these architectures at different scales. Moreover, it is now possible to tailor their biomechanical and topological properties while taking inspiration from the characteristics of the extracellular matrix, the three-dimensional scaffold in which cells proliferate, migrate, and differentiate. In such context, there is therefore a continuous quest for synthetic and nature-derived composite materials that must hold biocompatible, biodegradable, bioactive features and also be compatible with the envisioned fabrication strategy. The structure of the current review is intended to provide to both micro-engineers and cell biologists a comparative overview of the characteristics, advantages, and drawbacks of the major 3D printing techniques, the most promising biomaterials candidates, and the trade-offs that must be considered in order to replicate the properties of natural microenvironments. Full article

Solanum nigrum L. is one of the major medicinal plants used to treat cancer. However, the functional mechanism of S. nigrum L. extract is still unknown in spite of numerous studies on its active components. In this study, we probed the potential anticancer mechanism of the aqueous extract of S. nigrum L. (AESN) towards human breast cancer cell line MCF7. At a concentration of 10 g/L, AESN caused 43% cytotoxicity, inhibited the migration, and suppressed the activities of hexokinase and pyruvate kinase by about 30% and 40%, respectively, towards the MCF7 cells. RT²-PCR analysis of a panel of 89 caner-related genes identified 13 upregulated and eight downregulated genes (>2-folds) in MCF7 cells upon AESN treatment. Gene ontology (GO) and functional disease ontology (FunDO) analyses show that the antitumor function of S. nigrum L. involves multiple genes and these genes are shared across other diseases or disorders. Full article

X-ray computed tomography (CT) is widely used in clinical practice. The involved ionizing X-ray radiation, however, could increase cancer risk. Hence, the reduction of the radiation dose has been an important topic in recent years. Few-view CT image reconstruction is one of the main ways to minimize radiation dose and potentially allow a stationary CT architecture. In this paper, we propose a deep encoder-decoder adversarial reconstruction (DEAR) network for 3D CT image reconstruction from few-view data. Since the artifacts caused by few-view reconstruction appear in 3D instead of 2D geometry, a 3D deep network has a great potential for improving the image quality in a data driven fashion. More specifically, our proposed DEAR-3D network aims at reconstructing 3D volume directly from clinical 3D spiral cone-beam image data. DEAR is validated on a publicly available abdominal CT dataset prepared and authorized by Mayo Clinic. Compared with other 2D deep learning methods, the proposed DEAR-3D network can utilize 3D information to produce promising reconstruction results. Full article

Cell–cell communication plays a fundamental role in mediating corneal wound healing following injury or infection. Depending on the severity of the wound, regeneration of the cornea and the propensity for scar development are influenced by the acute resolution of the pro-fibrotic response mediated by closure of the wound via cellular and tissue contraction. Damage of the corneal epithelium, basement membrane, and anterior stroma following a superficial keratectomy is known to lead to significant provisional matrix deposition, including secretion of fibronectin and thrombospondin-1, as well as development of a corneal scar. In addition, corneal wounding has previously been shown to promote release of extracellular vesicles from the corneal epithelium, which, in addition to soluble factors, may play a role in promoting tissue regeneration. In this study, we report the development and characterization of a co-culture system of human corneal epithelial cells and corneal stromal fibroblasts cultured for 4 weeks to allow extracellular matrix deposition and tissue maturation. The secretion of provisional matrix components, as well as small and large extracellular vesicles, was apparent within the constructs, suggesting cell–cell communication between epithelial and stromal cell populations. Laminin-1β was highly expressed by the corneal epithelial layer with the presence of notable patches of basement membrane identified by transmission electron microscopy. Interestingly, we identified expression of collagen type III, fibronectin, and thrombospondin-1 along the epithelial–stromal interface similar to observations seen in vivo following a keratectomy, as well as expression of the myofibroblast marker, α-smooth muscle actin, within the stroma. Our results suggest that this corneal epithelial–stromal model may be useful in the study of the biochemical phenomena that occur during corneal wound healing. Full article

This is a literature teaching resource review for biologically inspired microfluidics courses or exploring the diverse applications of microfluidics. The structure is around key papers and model organisms. While courses gradually change over time, a focus remains on understanding how microfluidics has developed as well as what it can and cannot do for researchers. As a primary starting point, we cover micro-fluid mechanics principles and microfabrication of devices. A variety of applications are discussed using model prokaryotic and eukaryotic organisms from the set of bacteria (Escherichia coli), trypanosomes (Trypanosoma brucei), yeast (Saccharomyces cerevisiae), slime molds (Physarum polycephalum), worms (Caenorhabditis elegans), flies (Drosophila melangoster), plants (Arabidopsis thaliana), and mouse immune cells (Mus musculus). Other engineering and biochemical methods discussed include biomimetics, organ on a chip, inkjet, droplet microfluidics, biotic games, and diagnostics. While we have not yet reached the end-all lab on a chip, microfluidics can still be used effectively for specific applications. Full article

The rapidly growing interest on polyhydroxyalkanoates (PHA) processing for biomedical purposes is justified by the unique combinations of characteristics of this class of polymers in terms of biocompatibility, biodegradability, processing properties, and mechanical behavior, as well as by their great potential for sustainable production. This article aims at overviewing the most exploited processing approaches employed in the biomedical area to fabricate devices and other medical products based on PHA for experimental and commercial applications. For this purpose, physical and processing properties of PHA are discussed in relationship to the requirements of conventionally-employed processing techniques (e.g., solvent casting and melt-spinning), as well as more advanced fabrication approaches (i.e., electrospinning and additive manufacturing). Key scientific investigations published in literature regarding different aspects involved in the processing of PHA homo- and copolymers, such as poly(3-hydroxybutyrate), poly(3-hydroxybutyrate-co-3-hydroxyvalerate), and poly(3-hydroxybutyrate-co-3-hydroxyhexanoate), are critically reviewed. Full article

Thermoresponsive hydrogels showing biocompatibility and degradability have been under intense investigation for biomedical applications, especially hydrogels composed of hydrophilic poly(ethylene glycol) (PEG) and hydrophobic poly(lactic acid-co-glycolic acid) (PLGA) as first-line materials. Even though various aspects such as gelation behavior, degradation behavior, drug-release behavior, and composition effect have been studied for 20 years since the first report of these hydrogels, there are still many outputs on parameters affecting their gelation, structure, and application. In this review, the current trends of research on linear block copolymers composed of PEG and PLGA during the last 5 years (2014–2019) are summarized. In detail, this review stresses newly found parameters affecting thermoresponsive gelation, findings from structural analysis by simulation, small-angle neutron scattering (SANS), etc., progress in biomedical applications including drug delivery systems and regeneration medicine, and nanocomposites composed of block copolymers with PEG and PLGA and nanomaterials (laponite). Full article

Chlorantraniliprole (CAP) is a widely used insecticide in many areas due to its excellent insecticidal ability and mammalian safety, however, the removal of CAP has not been extensively studied. In this study, a bacterial strain GW13, which is capable of co-metabolizing CAP, was isolated from a vegetable field soil. The strain was identified as Pseudomonas sp. based on its physico-biochemical characteristics and 16S rRNA gene analysis. The bacterial strain GW13 could degrade CAP through co-metabolism, and glucose was the best additional carbon resource. In the presence of 1.0 g/L glucose, GW13 could co-metabolize over 80% of 200 mg/L CAP in 24 h. The degradation rate increased after 6 h and slowed again after 10 h. The GW13 genome analysis revealed many genes associated with metabolism, showing the degradation mechanism of GW13 from the genomic perspective. The EAWAG-BBD (Swiss Federal Institute of Aquatic Science and Technology Biocatalysis/Biodegradation Database) prediction results showed that the main pathway for CAP degradation is amide hydrolysis, which is consistent with many genes associated with amidase in the GW13 genome. This study may facilitate research on CAP biodegradation mechanisms in the environment. Full article

Some strains of Pseudomonas corrugata (Pco) and P. mediterranea (Pme) efficiently synthesize medium-chain-length polyhydroxyalkanoates elastomers (mcl-PHA) and extracellular products on related and unrelated carbon sources. Yield and composition are dependent on the strain, carbon source, fermentation process, and any additives. Selected Pco strains produce amorphous and sticky mcl-PHA, whereas strains of Pme produce, on high grade and partially refined biodiesel glycerol, a distinctive filmable PHA, very different from the conventional microbial mcl-PHA, suitable for making blends with polylactide acid. However, the yields still need to be improved and production costs lowered. An integrated process has been developed to recover intracellular mcl-PHA and extracellular bioactive molecules. Transcriptional regulation studies during PHA production contribute to understanding the metabolic potential of Pco and Pme strains. Data available suggest that pha biosynthesis genes and their regulations will be helpful to develop new, integrated strategies for cost-effective production. Full article

Artificial heart valves, used to replace diseased human heart valves, are life-saving medical devices. Currently, at the device development stage, new artificial valves are primarily assessed through time-consuming and expensive benchtop tests or animal implantation studies. Computational stress analysis using the finite element (FE) method presents an attractive alternative to physical testing. However, FE computational analysis requires a complex process of numeric modeling and simulation, as well as in-depth engineering expertise. In this proof of concept study, our objective was to develop machine learning (ML) techniques that can estimate the stress and deformation of a transcatheter aortic valve (TAV) from a given set of TAV leaflet design parameters. Two deep neural networks were developed and compared: the autoencoder-based ML-models and the direct ML-models. The ML-models were evaluated through Monte Carlo cross validation. From the results, both proposed deep neural networks could accurately estimate the deformed geometry of the TAV leaflets and the associated stress distributions within a second, with the direct ML-models (ML-model-d) having slightly larger errors. In conclusion, although this is a proof-of-concept study, the proposed ML approaches have demonstrated great potential to serve as a fast and reliable tool for future TAV design. Full article

Microfluidic-based technology attracts great interest in cell biology and medicine, in virtue of the ability to better mimic the in vivo cell microenvironment compared to conventional macroscale cell culture platforms. Recent Organs-on-chip (OoC) models allow to reproduce in vitro tissue and organ-level functions of living organs and systems. These models have been applied for the study of specific functions of the female reproductive tract, which is composed of several organs interconnected through intricate endocrine pathways and communication mechanisms. To date, a disease and toxicology study of this system has been difficult to perform. Thus, there is a compelling need to develop innovative platforms for the generation of disease model and for performing drug toxicity/screening in vitro studies. This review is focused on the analysis of recently published OoC models that recreate pathological and physiological characteristics of the female reproductive organs and tissues. These models aim to be used to assess changes in metabolic activity of the specific cell types and the effect of exposure to hormonal treatment or chemical substances on some aspects of reproduction and fertility. We examined these models in terms of device specifications, operating procedures, accuracy for studying the biochemical and functional activity of living tissues and the paracrine signalling that occurs within the different tissues. These models represent a powerful tool for understanding important diseases and syndromes affecting women all around the world. Immediate adoption of these models will allow to clarify diseases, causes and adverse events occurring during pregnancy such as pre-eclampsia, infertility or preterm birth, endometriosis and infertility. Full article

The use of lignocellulosic plant biomass as an alternative to fossil feedstocks for chemistry, energy and materials often involves an intense dry comminution step, for which the energy consumed can vary significantly according to the process parameters, the particle size targeted, and the properties of the biomass. Here we studied the fine milling of maritime pine bark in an impact-mill configuration and in an attrition-mill configuration. The properties of the resulting powders (particle size distribution, particle shape, specific surface area, agglomeration level) obtained in each configuration were compared in relation to process energy consumption. Results evidenced that the agglomeration phenomena drive milling efficiency and limit the possibilities for reaching ultrafine particles. Interestingly, impact loading proved more effective at breaking down coarse particles but tended to generate high agglomeration levels, whereas attrition milling led to less agglomeration and thus to finer particles. Full article

Mesenchymal stem cells have been widely implicated in tumour development and metastases. Moving from the use of two-dimensional (2D) models to three-dimensional (3D) to investigate this relationship is critical to facilitate more applicable and relevant research on the tumour microenvironment. We investigated the effects of altering glucose concentration and the source of foetal bovine serum (FBS) on the growth of two breast cancer cell lines (T47D and MDA-MB-231) and human bone marrow-derived mesenchymal stem cells (hBM-MSCs) to determine successful conditions to enable their co-culture in 3D tumour spheroid models. Subsequently, these 3D multi-cellular tumour spheroids were used to investigate the effect of hBM-MSCs on breast cancer cell invasiveness. Findings presented herein show that serum source had a statistically significant effect on two thirds of the growth parameters measured across all three cell lines, whereas glucose only had a statistically significant effect on 6%. It was determined that the optimum growth media composition for the co-culture of 3D hBM-MSCs and breast cancer cell line spheroids was 1 g/L glucose DMEM supplemented with 10% FBS from source A. Subsequent results demonstrated that co-culture of hBM-MSCs and MDA-MB-231 cells dramatically reduced invasiveness of both cell lines (F_(1,4) = 71.465, p = 0.001) when embedded into a matrix comprising of growth-factor reduced base membrane extract (BME) and collagen. Full article

Several nonlinear and anisotropic constitutive models have been proposed to describe the biomechanical properties of soft tissues, and reliably estimating the unknown parameters in these models using experimental data is an important step towards developing predictive capabilities. However, the effect of parameter estimation technique on the resulting biomechanical parameters remains under-analyzed. Standard off-the-shelf techniques can produce unreliable results where the parameters are not uniquely identified and can vary with the initial guess. In this study, a thorough analysis of parameter estimation techniques on the resulting properties for four multi-parameter invariant-based constitutive models is presented. It was found that linear transformations have no effect on parameter estimation for the presented cases, and nonlinear transforms are necessary for any improvement. A distinct focus is put on the issue of non-convergence, and we propose simple modifications that not only improve the speed of convergence but also avoid convergence to a wrong solution. The proposed modifications are straightforward to implement and can avoid severe problems in the biomechanical analysis. The results also show that including the fiber angle as an unknown in the parameter estimation makes it extremely challenging, where almost all of the formulations and models fail to converge to the true solution. Therefore, until this issue is resolved, a non-mechanical—such as optical—technique for determining the fiber angle is required in conjunction with the planar biaxial test for a robust biomechanical analysis. Full article

Flow processes and enzyme immobilization have gained much attention over the past few years in the field of biocatalytic process design. Downstream processes and enzyme stability can be immensely simplified and improved. In this work, we report the utilization of polymer network-entrapped enzymes and their applicability in flow processes. We focused on the superabsorber-based immobilization of an alcohol dehydrogenase (ADH) from Lactobacillus brevis and its application for a reduction of acetophenone. The applicability of this immobilization technique for a biotransformation running in a packed bed reactor was then demonstrated. Towards this end, the immobilized system was intensively studied, first in a batch mode, leading to >90% conversion within 24 h under optimized conditions. A subsequent transfer of this method into a flow process was conducted, resulting in very high initial conversions of up to 67% in such a continuously running process. Full article

In this study, we estimated and validated the pressure distribution profile between the residuum and two types of prosthetic sockets for transfemoral amputees by utilizing a finite element analysis. Correct shaping of the socket for an appropriate load distribution is a critical process in the design of lower-limb prosthesis sockets. The pressure distribution profile provides an understanding of the relationship between the socket design and the level of subject comfortability. Estimating the pressure profile is important, as it helps improve the prosthesis through an evaluation of the socket design before it undergoes the fabrication process. This study focused on utilizing a magnetic resonance imaging (MRI)-based three-dimensional (3D) model inside a predetermined finite element simulation. The simulation was predetermined by mimicking the actual socket-fitting environment. The results showed that the potential MRI-based 3D model simulation could be used as an estimation tool for a pressure distribution profile due to the high correlation coefficient value (R² > 0.8) calculated when the pressure profiles were compared to the experiment data. The simulation also showed that the pressure distribution in the proximal area was higher (~30%) than in the distal area of the prosthetic socket for every subject. The results of this study will be of tremendous interest for fabricators through the use of a finite element model as an alternative method for the prefabrication and evaluation of prosthetic sockets. In future prosthetic socket fabrications, less intervention will be required in the development of a socket, and the participation of the subject in the socket-fitting session will not be necessary. The results suggest that this study will contribute to expanding the development of an overall prefabrication evaluation system to allow healthcare providers and engineers to simulate the fit and comfort of transfemoral prosthetics. Full article

Hyaluronan (HA), a major component of the extracellular matrix, plays a key role in cell proliferation, growth, survival, polarization and differentiation. We investigated the optimization of a HA hydrogel scaffold for culture of human oral mucosal epithelial cells (OMECs) for potential application in limbal stem cell therapy. The effect of the optimized scaffold on OMEC cell sheet morphology, cell metabolic activity and expression of genes associated with stemness, adherence and cell damage was studied. The results indicate that HA hydrogels crosslinked with polyethylene glycol diacrylate (PEGDA) failed to support OMEC attachment and growth. However, HA hydrogel scaffolds dried for three days and coated with 1 mg/mL collagen IV produced a full OMEC sheet. Cell morphology was comparable to control after three weeks culture, maintaining 76% metabolic activity. Of apoptosis-related genes, the pro-apoptotic markers CASP3 and BAX2 were upregulated and downregulated, respectively, compared to control whereas the anti-apoptotic marker BCL2 was downregulated. The expression level of stemness genes ΔNp63α and ABCG2 was significantly higher than control. Genes associated with improved scar-less wound healing (integrin-αV) and protection of the ocular surface (cadherin-1) had ~3-fold increased expression. These data suggest that our optimized HA-hydrogel scaffold could enhance culture of OMEC cell sheets for use in ocular reconstruction. Full article

Bioprinting techniques can be used for the in vitro fabrication of functional complex bio-structures. Thus, extensive research is being carried on the use of various techniques for the development of 3D cellular structures. This article focuses on direct writing techniques commonly used for the fabrication of cell structures. Three different types of bioprinting techniques are depicted: Laser-based bioprinting, ink-jet bioprinting and extrusion bioprinting. Further on, a special reference is made to the use of the bioprinting techniques for the fabrication of 2D and 3D liver model structures and liver on chip platforms. The field of liver tissue engineering has been rapidly developed, and a wide range of materials can be used for building novel functional liver structures. The focus on liver is due to its importance as one of the most critical organs on which to test new pharmaceuticals, as it is involved in many metabolic and detoxification processes, and the toxicity of the liver is often the cause of drug rejection. Full article

Purpose: To evaluate the variation in the optical power achieved following soft contact lens eye fitting for spherical and cylindrical lenses with differing hydrogel material properties. Methods: Uniaxial tensile tests were performed on four hydrogel materials 77% water-content (w-c) hydrogel, 74% w-c blue silicone hydrogel, 74% w-c clear silicone hydrogel, and 64% w-c clear hydrogel (shortly referred to as H77p0, SiH74p5-blue, SiH74p5-clear, and H64p0-clear), under loading conditions that would be expected in vivo. Finite element models of the cornea and contact lens interaction were generated using spherical and cylindrical lenses with powers varying from −10 to +20 D; overall diameters of either 13.5, 14.0, or 14.5 mm; and with material properties matching those determined through experimental testing. Results: The moduli of elasticity for each of the tested hydrogel materials were 0.195 ± 0.027 MPa, 0.277 ± 0.019 MPa, 0.279 ± 0.01 MPa, and 0.457 ± 0.013 MPa for H77p0, SiH74p5-blue, SiH74p5-clear, and H64p0 respectively. The calculated values of effective power change (EPC) showed strong negative correlations with lens power. This was particularly apparent in the higher end of the lens power spectrum (over +5 D), where each of the materials demonstrated a highly linear reduction in EPC with increased lens power. Conclusions: Soft contact lenses composed of a stiffer hydrogel are far more resilient to changes in EPC across the lower end of the lens power spectrum (−10 to +5 D). Beyond this range, the material choice does not have a significant effect on the EPC. Full article

The rheology of high-cell density (HCD) cultures is an important parameter for its impact on mixing and sparging, process scale-up, and downstream unit operations in bioprocess development. In this work, time-dependent rheological properties of HCD Pseudomonas putida LS46 cultures were monitored for microbial polyhydroxyalkanoate (PHA) production. As the cell density of the fed-batch cultivation increased (0 to 25 g·L⁻¹ cell dry mass, CDM), the apparent viscosity increased nearly nine-fold throughout the fed-batch process. The medium behaved as a nearly Newtonian fluid at lower cell densities, and became increasingly shear-thinning as the cell density increased. However, shear-thickening behavior was observed at shearing rates of approximately 75 rad·s⁻¹ or higher, and its onset increased with viscosity of the sample. The supernatant, which contained up to 9 g·L⁻¹ soluble organic material, contributed more to the observed viscosity effect than did the presence of cells. Owing to this behavior, the oxygen transfer performance of the bioreactor, for otherwise constant operating conditions, was reduced by 50% over the cultivation time. This study has shown that the dynamic rheology of HCD cultures is an important engineering parameter that may impact the final outcome in PHA cultivations. Understanding and anticipating this behavior and its biochemical origins could be important for improving overall productivity, yield, process scalability, and the efficacy of downstream processing unit operations. Full article

The environmental impacts and high long-term costs of poor waste disposal have pushed the industry to realize the potential of turning this problem into an economic and sustainable initiative. Anaerobic digestion and the production of biogas can provide an efficient means of meeting several objectives concerning energy, environmental, and waste management policy. Biogas contains methane (60%) and carbon dioxide (40%) as its principal constituent. Excluding methane, other gasses contained in biogas are considered as contaminants. Removal of these impurities, especially carbon dioxide, will increase the biogas quality for further use. Integrating biological processes into the bio-refinery that effectively consume carbon dioxide will become increasingly important. Such process integration could significantly improve the sustainability of the overall bio-refinery process. The biogas upgrading by utilization of carbon dioxide rather than removal of it is a suitable strategy in this direction. The present work is a critical review that summarizes state-of-the-art technologies for biogas upgrading with particular attention to the emerging biological methanation processes. It also discusses the future perspectives for overcoming the challenges associated with upgradation. While biogas offers a good substitution for fossil fuels, it still not a perfect solution for global greenhouse gas emissions and further research still needs to be conducted. Full article

The liver is a key organ that can communicate with many other districts of the human body. In the last few decades, much interest has focused on the interaction between the liver and the gut microbiota, with their reciprocal influence on biosynthesis pathways and the integrity the intestinal epithelial barrier. Dysbiosis or liver disorders lead to0 epithelial barrier dysfunction, altering membrane permeability to toxins. Clinical and experimental evidence shows that the permeability hence the delivery of neurotoxins such as LPS, ammonia and salsolinol contribute to neurological disorders. These findings suggested multi-organ communication between the gut microbiota, the liver and the brain. With a view to in vitro modeling this liver-based multi-organ communication, we describe the latest advanced liver-on-a-chip devices and discuss the need for new organ-on-a-chip platforms for in vitro modeling the in vivo multi-organ connection pathways in physiological and pathological situations. Full article

Purpose: To investigate the impact of tissue derived biological particles on enzyme-mediated weakened corneas. Methods: Rabbit corneas were treated with enzymes to create an ex vivo ectatic model that simulated representative characteristics of keratoconus (KC). Porcine cornea, cartilage, and lymph node tissues were processed to remove most cellular components and cryomilled into microparticles. The KC corneas were cultured in medium containing the tissue-derived biological particles (TDP) overnight. The mechanical, thermal, ultrastructural changes, and gene expressions of corneal stromal cells were characterized to evaluate the effects of the TDP treatment. Results: The enzyme treatment significantly reduced corneal mechanics and thermal stability, and also disrupted the extracellular matrix ultrastructure. After culturing with TDP medium, the Young’s modulus of the modeled KC corneas increased by ~50%, comparable to normal cornea controls. Similarly, the thermal denaturation temperature of the corneas was restored. These findings also corresponded to a significant increase in collagen fibril density after TDP treatment. Furthermore, corneas cultured in TDP medium significantly downregulated expression of the pro-inflammatory gene Tnfα, and restored the expression of the key keratocyte markers Aldh, keratocan, and biglycan. Conclusions: Tissue-derived biological particles reinforce mechanical and thermal properties of corneal tissue in an ex vivo model of KC. Through this study, we demonstrate and characterize the previously unexplored impact of tissue-derived biological scaffolds on corneal biomechanics, thermal stability, and gene expression, presenting a potential new therapy for ocular disease. Full article

High cell density (HCD) fed-batch cultures are widely perceived as a requisite for high-productivity polyhydroxyalkanoate (PHA) cultivation processes. In this work, a reactive pulse feed strategy (based on real-time CO₂ or dissolved oxygen (DO) measurements as feedback variables) was used to control an oxygen-limited fed-batch process for improved productivity of medium chain length (mcl-) PHAs synthesized by Pseudomonas putida LS46. Despite the onset of oxygen limitation half-way through the process (14 h post inoculation), 28.8 ± 3.9 g L⁻¹ total biomass (with PHA content up to 61 ± 8% cell dry mass) was reliably achieved within 27 h using octanoic acid as the carbon source in a bench-scale (7 L) bioreactor operated under atmospheric conditions. This resulted in a final volumetric productivity of 0.66 ± 0.14 g L⁻¹ h⁻¹. Delivering carbon to the bioreactor as a continuous drip feed process (a proactive feeding strategy compared to pulse feeding) made little difference on the final volumetric productivity of 0.60 ± 0.04 g L⁻¹ h⁻¹. However, the drip feed strategy favored production of non-PHA residual biomass during the growth phase, while pulse feeding favored a higher rate of mcl-PHA synthesis and yield during the storage phase. Overall, it was shown that the inherent O₂-limitation brought about by HCD cultures can be used as a simple and effective control strategy for mcl-PHA synthesis from fatty acids. Furthermore, the pulse feed strategy appears to be a relatively easy and reliable method for rapid optimization of fed-batch processes, particularly when using toxic substrates like octanoic acid. Full article

Wilson Disease is a rare autosomal recessive liver disorder in humans. Although its clinical presentation and age of onset are highly variable, hallmarks include signs of liver disease, neurological features and so-called Kayser-Fleischer rings in the eyes of the patient. Hepatic copper accumulation leads to liver disease and eventually to liver cirrhosis. Treatment options include life-long copper chelation therapy and/or decrease in copper intake. Eventually liver transplantations are indicated. Although clinical outcome of liver transplantations is favorable, the lack of suitable donor livers hampers large numbers of transplantations. As an alternative, cell therapies with hepatocytes or liver stem cells are currently under investigation. Stem cell biology in relation to pets is in its infancy. Due to the specific population structure of dogs, canine copper toxicosis is frequently encountered in various dog breeds. Since the histology and clinical presentation resemble Wilson Disease, we combined genetics, gene-editing, and matrices-based stem cell cultures to develop a translational preclinical transplantation model for inherited copper toxicosis in dogs. Here we describe the roadmap followed, starting from the discovery of a causative copper toxicosis mutation in a specific dog breed and culminating in transplantation of genetically-engineered autologous liver stem cells. Full article

Biomass-derived coal or “biocoal” produced using a torrefaction process presents a carbon-neutral option of coal for power generation. While torrefaction delivers a carbon content and hydrophobicity comparable to coal, it lowers its density and creates material handling, storage, and transportation challenges. Densification into briquettes would help mitigate these challenges. However, the torrefied biomass is difficult to densify and may require the use of binders, which are expensive and can be incompatible with respect to material and emissions. A cost-effective approach to utilize lignin in-situ of the biomass to promote binding during densification was demonstrated using a pilot-scale briquetter unit during this study. Lignin, a cross-linked polymer, tends to break down and lose its binding ability under high-temperature conditions of torrefaction. In this paper, we investigated the use of a lightly torrefied material as a binder―LTM (biomass torrefied in the transition region of non-reactive and reactive temperature ranges of torrefaction). When mixed with torrefied biomass and densified together under suitable moisture and temperature conditions, the lignin is shown to mobilize and provide binding to the briquettes. The results showed that briquettes produced using LTM as binder and 10% to 11% moisture provided in-situ binding, improved density and durability, and produced hydrophobic briquettes. Full article