Tunable Fungal Monofilaments from Food Waste for Textile Applications.

A fungal biorefinery is presented to valorize food waste to fungal monofilaments with tunable properties for different textile applications. Rhizopus delemar is successfully grown on bread waste and the fibrous cell wall is isolated. A spinnable hydrogel is produced from cell wall by protonation of amino groups of chitosan followed by homogenization and concentration. Fungal hydrogel is wet spun to form fungal monofilaments which underwent post-treatments to tune the properties. The highest tensile strength of untreated monofilaments is 65 MPa (and 4% elongation at break). The overall highest tensile strength of 140.9 MPa, is achieved by water post-treatment. Moreover, post-treatment with 3% glycerol resulted in the highest elongation % at break, i.e., 14%. The uniformity of the monofilaments also increased after the post-treatments. The obtained monofilaments are compared with commercial fibers using Ashby's plots and potential applications are discussed. The wet spun monofilaments are located in the category of natural fibers in Ashby's plots. After water and glycerol treatments, the properties shifted toward metals and elastomers, respectively. The compatibility of the monofilaments with human skin cells is supported by a biocompatibility assay. These findings demonstrate fungal monofilaments with tunable properties fitting a wide range of sustainable textiles applications.

Development of an Open-Source Injection Mold Monitoring System.

In the highly competitive injection molding industry, the ability to effectively collect information from various sensors installed in molds and machines is of the utmost relevance, enabling the development of data-based Industry 4.0 algorithms. In this work, an alternative to commercially available monitoring systems used in the industry was developed and tested in the scope of the TOOLING 4G project. The novelty of this system is its affordability, simplicity, real-time data acquisition and display in an intuitive Graphical User Interface (GUI), while being open-source firmware and software-based. These characteristics, and their combinations have been present in previous works, but, to the authors' knowledge, not all of them simultaneously. The system used an Arduino microcontroller-based data acquisition module that can be connected to any computer via a USB port. Software was developed, including a GUI, prepared to receive data from both the Arduino module and a second module. In the current state of development, data corresponding to a maximum of six sensors can be visualized, at a rate of 10 Hz, and recorded for later usage. These capabilities were verified under real-world conditions for monitoring an injection mold with the objective of creating the basis of a platform to deploy predictive maintenance. Mold temperature, cavity pressure, 3-axis acceleration, and extraction force data showed the system can successfully monitor the mold and allowed the clear distinction between normal and abnormal operating patterns.

Turning food waste to antibacterial and biocompatible fungal chitin/chitosan monofilaments.

Here, cell wall of a zygomycete fungus, Rhizopus delemar, grown on bread waste was wet spun into monofilaments. Using the whole cell wall material omits the common chitosan isolation and purification steps and leads to higher material utilization. The fungal cell wall contained 36.9% and 19.7% chitosan and chitin, respectively. Solid state NMR of the fungal cell wall material confirmed the presence of chitosan, chitin, and other carbohydrates. Hydrogels were prepared by ultrafine grinding of the cell wall, followed by addition of lactic acid to protonate the amino groups of chitosan, and subsequently wet spun into monofilaments. The monofilament inhibited the growth of Bacillus megaterium (Gram+ bacterium) and Escherichia coli (Gram- bacterium) significantly (92.2% and 99.7% respectively). Cytotoxicity was evaluated using an in vitro assay with human dermal fibroblasts, indicating no toxic inducement from exposure of the monofilaments. The antimicrobial and biocompatible fungal monofilaments, open new avenues for sustainable biomedical textiles from abundant food waste.

Multiscale Sensing of Bone-Implant Loosening for Multifunctional Smart Bone Implants: Using Capacitive Technologies for Precision Controllability.

The world population growth and average life expectancy rise have increased the number of people suffering from non-communicable diseases, namely osteoarthritis, a disorder that causes a significant increase in the years lived with disability. Many people who suffer from osteoarthritis undergo replacement surgery. Despite the relatively high success rate, around 10% of patients require revision surgeries, mostly because existing implant technologies lack sensing devices capable of monitoring the bone-implant interface. Among the several monitoring methodologies already proposed as substitutes for traditional imaging methods, cosurface capacitive sensing systems hold the potential to monitor the bone-implant fixation states, a mandatory capability for long-term implant survival. A multifaceted study is offered here, which covers research on the following points: (1) the ability of a cosurface capacitor network to effectively monitor bone loosening in extended peri-implant regions and according to different stimulation frequencies; (2) the ability of these capacitive architectures to provide effective sensing in interfaces with hydroxyapatite-based layers; (3) the ability to control the operation of cosurface capacitive networks using extracorporeal informatic systems. In vitro tests were performed using a web-based network sensor composed of striped and interdigitated capacitive sensors. Hydroxyapatite-based layers have a minor effect on determining the fixation states; the effective operation of a sensor network-based solution communicating through a web server hosted on Raspberry Pi was shown. Previous studies highlight the inability of current bone-implant fixation monitoring methods to significantly reduce the number of revision surgeries, as well as promising results of capacitive sensing systems to monitor micro-scale and macro-scale bone-interface states. In this study, we found that extracorporeal informatic systems enable continuous patient monitoring using cosurface capacitive networks with or without hydroxyapatite-based layers. Findings presented here represent significant advancements toward the design of future multifunctional smart implants.

Capacitive interdigitated system of high osteoinductive/conductive performance for personalized acting-sensing implants.

Replacement orthopedic surgeries are among the most common surgeries worldwide, but clinically used passive implants cannot prevent failure rates and inherent revision arthroplasties. Optimized non-instrumented implants, resorting to preclinically tested bioactive coatings, improve initial osseointegration but lack long-term personalized actuation on the bone-implant interface. Novel bioelectronic devices comprising biophysical stimulators and sensing systems are thus emerging, aiming for long-term control of peri-implant bone growth through biointerface monitoring. These acting-sensing dual systems require high frequency (HF) operations able to stimulate osteoinduction/osteoconduction, including matrix maturation and mineralization. A sensing-compatible capacitive stimulator of thin interdigitated electrodes and delivering an electrical 60 kHz HF stimulation, 30 min/day, is here shown to promote osteoconduction in pre-osteoblasts and osteoinduction in human adipose-derived mesenchymal stem cells (hASCs). HF stimulation through this capacitive interdigitated system had significant effects on osteoblasts' collagen-I synthesis, matrix, and mineral deposition. A proteomic analysis of microvesicles released from electrically-stimulated osteoblasts revealed regulation of osteodifferentiation and mineralization-related proteins (e.g. Tgfb3, Ttyh3, Itih1, Aldh1a1). Proteomics data are available via ProteomeXchange with the identifier PXD028551. Further, under HF stimulation, hASCs exhibited higher osteogenic commitment and enhanced hydroxyapatite deposition. These promising osteoinductive/conductive capacitive stimulators will integrate novel bioelectronic implants able to monitor the bone-implant interface and deliver personalized stimulation to peri-implant tissues.

Starch and protein recovery from brewer's spent grain using hydrothermal pretreatment and their conversion to edible filamentous fungi - A brewery biorefinery concept.

This study aimed at recovering a highly concentrated starch and protein stream from the brewer's spent grain (BSG). The effect of pretreatment temperature and retention time on the solubilization of starch and protein; and the generation of fermentation inhibitors were studied. Then, the application of recovered streams for fungal cultivation was evaluated using different edible fungi Aspergillus oryzae, Neurospora intermedia, and Rhizopus delemar. The hydrothermal pretreatment resulted in the highest solubilized starch concentration, 43 g/L, corresponding to 83% solubilization of initial BSG starch content. The highest protein concentration was 27 g/L (48% solubilization of initial BSG protein content). Cultivation with Neurospora intermedia on the recovered streams from the two best pretreatment conditions, 140 ℃ for 4 h and 180 ℃ for 30 min, resulted in pure fungal biomass with the highest protein content 59.62% and 50.42% w/w, respectively. Finally, a brewery biorefinery was proposed for the valorization of BSG.

Production of polyhydroxyalkanoates (PHAs) by Bacillus megaterium using food waste acidogenic fermentation-derived volatile fatty acids.

High production costs still hamper fast expansion of commercial production of polyhydroxyalkanoates (PHAs). This problem is greatly related to the cultivation medium which accounts for up to 50% of the whole process costs. The aim of this research work was to evaluate the potential of using volatile fatty acids (VFAs), derived from acidogenic fermentation of food waste, as inexpensive carbon sources for the production of PHAs through bacterial cultivation. Bacillus megaterium could assimilate glucose, acetic acid, butyric acid, and caproic acid as single carbon sources in synthetic medium with maximum PHAs production yields of 9-11%, on a cell dry weight basis. Single carbon sources were then replaced by a mixture of synthetic VFAs and by a VFAs-rich stream from the acidogenic fermentation of food waste. After 72 h of cultivation, the VFAs were almost fully consumed by the bacterium in both media and PHAs production yields of 9-10%, on cell dry weight basis, were obtained. The usage of VFAs mixture was found to be beneficial for the bacterial growth that tackled the inhibition of propionic acid, iso-butyric acid, and valeric acid when these volatile fatty acids were used as single carbon sources. The extracted PHAs were revealed to be polyhydroxybutyrate (PHB) by characterization methods of Fourier-transform infrared spectroscopy (FTIR) and differential scanning calorimetry (DSC). The obtained results proved the possibility of using VFAs from acidogenic fermentation of food waste as a cheap substrate to reduce the cost of PHAs production.

Towards an effective sensing technology to monitor micro-scale interface loosening of bioelectronic implants.

Instrumented implants are being developed with a radically innovative design to significantly reduce revision surgeries. Although bone replacements are among the most prevalent surgeries performed worldwide, implant failure rate usually surpasses 10%. High sophisticated multifunctional bioelectronic implants are being researched to incorporate cosurface capacitive architectures with ability to deliver personalized electric stimuli to peri-implant target tissues. However, the ability of these architectures to detect bone-implant interface states has never been explored. Moreover, although more than forty technologies were already proposed to detect implant loosening, none is able to ensure effective monitoring of the bone-implant debonding, mainly during the early stages of loosening. This work shows, for the first time, that cosurface capacitive sensors are a promising technology to provide an effective monitoring of bone-implant interfaces during the daily living of patients. Indeed, in vitro experimental tests and simulation with computational models highlight that both striped and circular capacitive architectures are able to detect micro-scale and macro-scale interface bonding, debonding or loosening, mainly when bonding is weakening or loosening is occurring. The proposed cosurface technologies hold potential to implement highly effective and personalized sensing systems such that the performance of multifunctional bioelectronic implants can be strongly improved. Findings were reported open a new research line on sensing technologies for bioelectronic implants, which may conduct to great impacts in the coming years.

Production of L-carnitine-enriched edible filamentous fungal biomass through submerged cultivation.

The edible filamentous fungi are hot candidate for future supply of functional food and feed with e.g. protein, essential amino acids, and compounds with immunostimulant activity. L-carnitine that plays a crucial role in energy metabolism represents a functional compound normally produced by Zygomycetes filamentous fungus Rhizopus oligosporus in solid-state fermentation. The present study provides the first insights on production of L-carnitine-enriched edible fungal biomass through submerged cultivation of several Ascomycetes and Zygomycetes including Aspergillus oryzae, Neurospora intermedia, Rhizopus oryzae, and Rhizopus oligosporus. A. oryzae with 3 mg L-carnitine yield per gram of fungal biomass, indicates great potential on production of this bioactive compound which is remarkably higher than the other tested fungi in this work and also previous studies. In addition to fungal strain, other factors such as cultivation time and presence of yeast extract were found to play a role. Further studies on submerged growth optimization of A. oryzae in both high-quality recipes and in medium based on low-value substrates are proposed in order to clarify its potential for production of L-carnitine-enriched fungal biomass.

Bioprocessing strategies to increase the protein fraction of Rhizopus oryzae biomass using fish industry sidestreams.

The sidestreams produced during fish processing end in a separation tank where the resulting fractions follow biogas production or wastewater treatment. These streams can alternatively be used for production of protein-rich fungal biomass for e.g. fish feed applications, a product in increasing demand. These streams and upper streams originated during fish processing were used in this study for production of biomass using the edible filamentous fungus Rhizopus oryzae. The COD of the streams varied between 11 and 54 kg/m3 and, after fungal conversion of organic matter into protein-rich biomass and separation, a reduction of 34-69% was achieved. The stream origin had an effect on the final production and composition of the fungal biomass: 480 kg of biomass containing 33% protein per ton of COD were produced after cultivation in the separation tank streams, while 220 kg of biomass containing 62% protein per ton of COD were produced in upper sidestreams with lower amounts of suspended solids. Changing the initial pH (6.1-6.5) to 5.0 had a negative influence on the amount of biomass produced while medium supplementation had no influence. Thus, fish processing sidestreams can be diverted from biogas production and wastewater treatment to the production of protein-rich biomass for feed applications.

Improving the economy of lignocellulose-based biorefineries with organosolv pretreatment.

Lignocellulose-based processes for production of value-added products still face bottlenecks to attain feasibility. The key might lie on the biorefining of all lignocellulose main polymers, that is, cellulose, hemicellulose and lignin. Lignin, considered an impediment in the access of cellulose and normally considered for energy recovery purposes, can give a higher contribution towards profitability of lignocellulosic biorefineries. Organosolv pretreatment allows selective fractionation of lignocellulose into separate cellulose-, hemicellulose- and lignin-rich streams. Ethanol organosolv and wood substrates dominated the research studies, while a wide range of substrates need definition on the most suitable organosolv pretreatment systems. Techno-economic and environmental analyses of organosolv-based processes as well as proper valorization strategies of the hemicellulose-rich fraction are still scarce. In view of dominance of ethanol organosolv with high delignification yields and high-purity of the recovered cellulose-rich fractions, close R & D collaboration with 1st generation ethanol plants might boost commercialization.

Recycling strategies for polyhydroxyalkanoate-based waste materials: An overview.

The plastics market is dominated by fossil-based polymers, but their gradual replacement by bioplastics (e.g., polyhydroxyalkanoates) is occurring. However, recycling strategies need to be developed to truly unveil the impact of bioplastics on waste accumulation. This review provides a state of the art of recycling strategies investigated for polyhydroxyalkanoate-based polymers and proposes future research avenues. Research on mechanical and chemical recycling is dominated by the use of extrusion and pyrolysis, respectively, while that on biodegradation of polyhydroxyalkanoates is related to soil and aquatic samples, and to anaerobic digestion towards biogas production. Research gaps exist in the relationships between polymer composition and ease of use of all recycling strategies investigated. This is of utmost importance since it will influence the need for separation at the source. Therefore, research emphasis needs to be given to the area to follow the continuous improvement of the process economics towards widespread commercial production of polyhydroxyalkanoates.

Altering the Course of Technologies to Monitor Loosening States of Endoprosthetic Implants.

Musculoskeletal disorders are becoming an ever-growing societal burden and, as a result, millions of bone replacements surgeries are performed per year worldwide. Despite total joint replacements being recognized among the most successful surgeries of the last century, implant failure rates exceeding 10% are still reported. These numbers highlight the necessity of technologies to provide an accurate monitoring of the bone-implant interface state. This study provides a detailed review of the most relevant methodologies and technologies already proposed to monitor the loosening states of endoprosthetic implants, as well as their performance and experimental validation. A total of forty-two papers describing both intracorporeal and extracorporeal technologies for cemented or cementless fixation were thoroughly analyzed. Thirty-eight technologies were identified, which are categorized into five methodologies: vibrometric, acoustic, bioelectric impedance, magnetic induction, and strain. Research efforts were mainly focused on vibrometric and acoustic technologies. Differently, approaches based on bioelectric impedance, magnetic induction and strain have been less explored. Although most technologies are noninvasive and are able to monitor different loosening stages of endoprosthetic implants, they are not able to provide effective monitoring during daily living of patients.

Mycoprotein: environmental impact and health aspects.

The term mycoprotein refers to the protein-rich food made of filamentous fungal biomass that can be consumed as an alternative to meat. In this paper, the impact caused by the substitution of animal-origin meat in the human diet for mycoprotein on the health and the environment is reviewed. Presently, mycoprotein can be found in the supermarkets of developed countries in several forms (e.g. sausages and patties). Expansion to other markets depends on the reduction of the costs. Although scarce, the results of life cycle analyses of mycoprotein agree that this meat substitute causes an environmental impact similar to chicken and pork. In this context, the use of inexpensive agro-industrial residues as substrate for mycoprotein production has been investigated. This strategy is believed to reduce the costs involved in the fungal cultivation and lower the environmental impact of both the mycoprotein and the food industry. Moreover, several positive effects in health have been associated with the substitution of meat for mycoprotein, including improvements in blood cholesterol concentration and glycemic response. Mycoprotein has found a place in the market, but questions regarding the consumer's experience on the sensory and health aspects are still being investigated.

Post-treatment of Fungal Biomass to Enhance Pigment Production.

A new post-treatment method of fungal biomass after fermentation is revealed. The post-treatment strategy was utilized to produce pigments as an additional valuable metabolite. Post-treatment included incubation at 95% relative humidity where the effects of harvesting time, light, and temperature were studied. Pigment-producing edible filamentous fungus Neurospora intermedia cultivated on ethanol plant residuals produced 4 g/L ethanol and 5 g/L fungal biomass. Harvesting the pale biomass after 48 h submerged cultivation compared to 24 h or 72 h increased pigmentation in the post-treatment step with 35% and 48%, respectively. The highest pigment content produced, 1.4 mg/g dry fungal biomass, was obtained from washed biomass treated in light at 35 °C whereof the major impact on pigmentation was from washed biomass. Moreover, post-treated biomass contained 50% (w/w) crude protein. The post-treatment strategy successfully adds pigments to pre-obtained biomass. The pigmented fungal biomass can be considered for animal feed applications for domestic animals.

Capacitive technologies for highly controlled and personalized electrical stimulation by implantable biomedical systems.

Cosurface electrode architectures are able to deliver personalized electric stimuli to target tissues. As such, this technology holds potential for a variety of innovative biomedical devices. However, to date, no detailed analyses have been conducted to evaluate the impact of stimulator architecture and geometry on stimuli features. This work characterizes, for the first time, the electric stimuli delivered to bone cellular tissues during in vitro experiments, when using three capacitive architectures: stripped, interdigitated and circular patterns. Computational models are presented that predict the influence of cell confluence, cosurface architecture, electrodes geometry, gap size between electrodes and power excitation on the stimuli delivered to cellular layers. The results demonstrate that these stimulators are able to deliver osteoconductive stimuli. Significant differences in stimuli distributions were observed for different stimulator designs and different external excitations. The thickness specification was found to be of utmost importance. In vitro experiments using an osteoblastic cell line highlight that cosurface stimulation at a low frequency can enhance osteoconductive responses, with some electrode-specific differences being found. A major feature of this type of work is that it enables future detailed analyses of stimuli distribution throughout more complex biological structures, such as tissues and organs, towards sophisticated biodevice personalization.

Changes in carbon footprint when integrating production of filamentous fungi in 1st generation ethanol plants.

Integrating the cultivation of edible filamentous fungi in the thin stillage from ethanol production is presently being considered. This integration can increase the ethanol yield while simultaneously producing a new value-added protein-rich biomass that can be used for animal feed. This study uses life cycle assessment to determine the change in greenhouse gas (GHG) emissions when integrating the cultivation of filamentous fungi in ethanol production. The result shows that the integration performs better than the current scenario when the fungal biomass is used as cattle feed for system expansion and when energy allocation is used. It performs worse if the biomass is used as fish feed. Hence, integrating the cultivation of filamentous fungi in 1st generation ethanol plants combined with proper use of the fungi can lead to a reduction of GHG emissions which, considering the number of existing ethanol plants, can have a significant global impact.

Filamentous ascomycetes fungi as a source of natural pigments.

Filamentous fungi, including the ascomycetes Monascus, Fusarium, Penicillium and Neurospora, are being explored as novel sources of natural pigments with biological functionality for food, feed and cosmetic applications. Such edible fungi can be used in biorefineries for the production of ethanol, animal feed and pigments from waste sources. The present review gathers insights on fungal pigment production covering biosynthetic pathways and stimulatory factors (oxidative stress, light, pH, nitrogen and carbon sources, temperature, co-factors, surfactants, oxygen, tricarboxylic acid intermediates and morphology) in addition to pigment extraction, analysis and identification methods. Pigmentation is commonly regarded as the output of secondary protective mechanisms against oxidative stress and light. Although several studies have examined pigmentation in Monascus spp., research gaps exist in the investigation of interactions among factors as well as process development on larger scales under submerged and solid-state fermentation. Currently, research on pigmentation in Neurospora spp. is at its infancy, but the increasing interest for biorefineries shows potential for booming research in this area.

Mild-temperature dilute acid pretreatment for integration of first and second generation ethanol processes.

The use of hot-water (100°C) from the 1st generation ethanol plants for mild-temperature lignocellulose pretreatment can possibly cut down the operational (energy) cost of 2nd generation ethanol process, in an integrated model. Dilute-sulfuric and -phosphoric acid pretreatment at 100°C was carried out for wheat bran and whole-stillage fibers. Pretreatment time and acid type influenced the release of sugars from wheat bran, while acid-concentration was found significant for whole-stillage fibers. Pretreatment led up-to 300% improvement in the glucose yield compared to only-enzymatically treated substrates. The pretreated substrates were 191-344% and 115-300% richer in lignin and glucan, respectively. Fermentation using Neurospora intermedia, showed 81% and 91% ethanol yields from wheat bran and stillage-fibers, respectively. Sawdust proved to be a highly recalcitrant substrate for mild-temperature pretreatment with only 22% glucose yield. Both wheat bran and whole-stillage are potential substrates for pretreatment using waste heat from the 1st generation process for 2nd generation ethanol.