Conventional treatment for esophageal defects involves surgical removal of the defect area and implant conduit tissues. There exist morbidities and mortalities associated with the treatment including fistula and leakage leading to compromise in quality of life. The aim of this study was to optimize a method for complete decellularization of rat esophagus and to solubilize the decellularized extracellular matrix (dECM) proteins to evaluate in vitro properties for scaffold fabrication. For decellularization, rat esophagi were decellularized using 3-[(3-cholamidopropyl) dimethylammonio]-1-propanesulfonate (CHAPS) and sodium dodecyl sulfate (SDS) buffers for 6 h and overnight, respectively. Post decellularization, the tissue was characterized for DNA, glycosaminoglycans, and elastin quantification; H&E and Masson's trichrome staining; scanning electron microscopy; and SDS-PAGE to evaluate the quantity and quality of the obtained dECM. DNA quantification and histological analysis revealed complete decellularization, while the retention of sGAGs and elastin showed the presence of extracellular proteins in the tissue. The SEM analysis revealed proper orientation of the extracellular matrix and significant proteins were retained in the dECM, which will enhance the regenerative potential. The decellularized tissues were biocompatible, exhibited no toxicity and were also soluble, which can be adapted for scaffold fabrication.
Spent mushroom substrate (SMS), the main by-product of mushroom cultivation, is a source of sugars that can be released by saccharification. This work aimed at investigating the enzymatic saccharification of the polysaccharides of the SMS of shiitake (Lentinula edodes) and oyster mushroom (Pleurotus ostreatus) and exploring the lignin extraction from the saccharification residues. First, analytical enzymatic saccharification (AES) with a cellulase cocktail and an experimental hemicellulase-rich preparation was applied. AES revealed higher digestibility of both polysaccharides for shiitake SMS than for oyster mushroom SMS. Using the cellulase cocktail, shiitake SMS resulted in a digestibility above 80% and 70% (w/w) for cellulose and xylan, respectively, while the maximum values for oyster mushroom SMS were 52% and 32% (w/w). The experimental enzyme preparation resulted in lower cellulose digestibility and higher xylan digestibility. Still, the saccharification trend between the two SMS types remained unchanged. To enhance the enzymatic saccharification of oyster mushroom SMS, hydrothermal treatment was applied. The treatment improved the enzymatic digestibility of cellulose by up to 84%. A validation experiment at larger scale showed that hydrothermally treated oyster mushroom SMS had a comparable overall conversion with non-treated shiitake SMS. Following a biorefinery strategy, lignin was extracted from the residues of the preparative enzymatic saccharification using the green solvent γ-valerolactone under different temperatures and holding times. The extracted product contained 98.8% lignin and did not contain cellulose or xylan. The results of this study provide the grounds for biorefinery processes enabling recovery of bioactive compounds, fermentable sugars, and high-quality lignin from SMS.
Anaerobic digestion (AD) is a sustainable technology that converts organic waste into renewable energy while reducing greenhouse gas emissions. Recent studies suggest that adding CO₂ to the AD process can improve methane production through different mechanisms. This review examines four key ways CO₂ supplementation can enhance methane yield: (1) direct conversion of CO₂ into acetate by homoacetogens, (2) direct methanation of CO₂ by hydrogenotrophic methanogens, (3) improved breakdown of organic material due to higher enzyme activity, and (4) better digester conditions through pH regulation and reduced ammonia toxicity. By analyzing microbial interactions and process improvements, this paper highlights knowledge gaps and the need for further research to optimize CO₂ addition in different operational settings. These findings are expected to contribute to the development of cost-effective and efficient AD systems that support energy recovery and environmental sustainability.
Seventeen halotolerant bacteria were isolated from the Muthupettai mangroves, Tamil Nadu, India, with eight exhibiting protease production. The most potent isolate, VITGS4, identified as Streptomyces sp. via polyphasic taxonomy, yielded 470 U mL-1. Response surface methodology (RSM) optimized protease production by Box-Behnken Design (BBD) using casein (5.5% w/v), pH 7.5, and 9.5 days incubation, achieving 282 U mL-1. The recovered protease was partially purified through acetone precipitation (50% acetone), followed by dialysis, and its purity was estimated through HPLC (high pressure liquid chromatography). Enzyme kinetics revealed a Km of 0.347 µM, a Vo of 0.464 µM min-1, a Vmax of 3.167 µM min-1, and a Kcat of 0.0002 min-1. The enzyme was identified as a halo-thermo-alkaline serine protease, optimally active at pH 8 and 45°C, with activity significantly inhibited by Pb2+ and Hg2+ and enhanced by Zn2+ (95%). Notably, PMSF strongly inhibited protease activity, indicating a serine protease. This protease was successfully employed to recover 726 mg of silver slurry (537 µg mL-1 silver) from X-ray films. Scanning electron microscopy (SEM) and energy-dispersive X-ray spectroscopy (EDX) confirmed the presence of silver (2.2% in the analyzed region), while zeta potential (-26.35 mV) and hydrodynamic diameter (89.94 nm) analyses indicated stable silver nanoparticles. These results demonstrate the potential of marine actinobacteria-derived proteases for efficient silver recovery, offering promising applications in therapeutic and industrial fields.
Fiberbanks are organic-rich sediment deposits in aquatic environments, primarily formed through historical pulp and paper mill activities. These deposits consist of wood-derived fibrous materials and are contaminated with potentially toxic elements (PTEs) such as vanadium, chromium, cobalt, nickel, copper, zinc, arsenic, cadmium, and lead. The leaching of these contaminants into surrounding waters poses significant environmental and health risks, impacting aquatic ecosystems and potentially entering the food chain. Effective remediation of fiberbanks is crucial, particularly in Sweden and other regions with extensive wood-pulping industries. This study aims to evaluate the bioaccumulation capacities of 26 native Swedish white-rot fungi (WRF) species for the remediation of PTEs in fiberbank material. Fiberbank samples were collected from Sundsvall's Bay in the Baltic Sea, while the fungal species were isolated from boreal forests in Västernorrland, Sweden. The fungi were cultured on Hagem agar medium with sterilized fiberbank material as the substrate. After two months, fungal biomass was analyzed for PTE uptake using inductively coupled plasma-mass spectrometry (ICP-MS). The results revealed significant variability (p < 0.001) in PTE uptake among fungal species. Phlebia tremellosa consistently demonstrated the highest bioconcentration factors for analyzed elements, with values for V (0.39), Cr (0.10), Co (1.81), Cu (1.54), Pb (1.65), Ni (1.28), As (0.83), Zn (3.61), and Cd (5.56). Other species, including Laetiporus sulphureus (0.09-4.78), Hymenochaete tabacina (0.08-4.52), and Diplomitoporus crustulinus (0.08-4.48), also exhibited significant bioremediation potential. These findings highlight the potential of native WRF species for PTEs remediation in fiberbanks and provide a foundation for mycoremediation strategies in contaminated environments.
Osteoarthritis is a prevalent degenerative joint disease characterized by cartilage degradation, synovial inflammation, and subchondral bone alterations, leading to chronic pain and joint dysfunction. Conventional treatments provide symptomatic relief but fail to halt disease progression. Recent advancements in biomaterials, molecular signaling modulation, and gene-editing technologies offer promising therapeutic strategies. This review explores key molecular pathways implicated in osteoarthritis, including fibroblast growth factor, phosphoinositide 3-kinase/Akt, and bone morphogenetic protein signaling, highlighting their roles in chondrocyte survival, extracellular matrix remodeling, and inflammation. Biomaterial-based interventions such as hydrogels, nanoparticles, and chitosan-based scaffolds have demonstrated potential in enhancing cartilage regeneration and targeted drug delivery. Furthermore, CRISPR/Cas9 gene editing holds promise in modifying osteoarthritis-related genes to restore cartilage integrity. The integration of regenerative biomaterials with precision medicine and molecular therapies represents a novel approach for mitigating osteoarthritis progression. Future research should focus on optimizing biomaterial properties, refining gene-editing efficiency, and developing personalized therapeutic strategies. The convergence of bioengineering and molecular science offers new hope for improving joint function and patient quality of life in osteoarthritis management.
Taylor & Francis journal Bioengineered has been targeted by paper mills. Our goal is to identify problematic articles published in Bioengineered during the period 2010 to 2024. Dimensions was used to search for articles that contained the terms 'mouse' OR 'mice' OR 'rat' OR 'rats' in title or abstract, published in Bioengineered between January 1st 2010 to December 31st 2024. All articles were assessed by eye and by using software to detect inappropriate image duplication and manipulation. An article was classified as problematic if it contained inappropriate image duplication or manipulation or had been previously retracted. Problematic articles were reported on PubPeer by the authors if they had not been reported previously. All included articles were assessed for post-publication editorial decisions. We have excluded all articles published in 2024 from further analysis, as these were all retraction notices. We assessed the remaining 878 articles, of which 226 (25.7%) were identified as problematic, of which 35 had been previously retracted. One retracted article was later de-retracted. One article received a correction. None of the included articles received an expression of concern or the Taylor & Francis 'under investigation' pop-up. Taylor & Francis' lack of visible editorial action has left the scientific community vulnerable to reading and citing hundreds of problematic articles published in Bioengineered. To uphold scientific integrity, Taylor & Francis should use the findings of this study as a starting point to systematically identify all compromised articles in Bioengineered and take appropriate editorial action.
Substituting waste-derived Volatile Fatty Acids (VFAs) with their conventionally applied fossil-derived counterparts in a spectrum of industrial applications necessitates its proper fractionation into individual acids. This study explored a multi-stage batch adsorption approach for fractionating acidogenic fermentation VFAs effluents from food waste (FW) and chicken manure (CKM) using Diaion HP-20 and activated charcoal. Initial screening at different washing conditions and pH (3.5 and 6.5) revealed the unwashed granular-activated charcoal (GAC-Unwashed) and milli-Q water-washed Diaion (DI-MQ Washed) as the most promising candidates for VFA fractionation of a synthetic VFA mixture at 4 gL-1. At pH 3.5 (97%). While at pH 6.5 (>pKa), GAC-Unwashed selectively targeted VA (79%) and CA (100%). Fractionating VFAs from FW and CKM were conducted in a two-stage adsorption process with optimal results being achieved using GAC-Unwashed at FW initial pH (5.3) and DI-MQ Washed at pH below CKM pKa (3.5), respectively. The first adsorption stage primarily adsorbed higher molecular weight (MW) VFAs (FW:99.1% CA, CKM:72.9% butyric acid (BA)) with a minor quantity of lower ones (FW:56.5% BA, CKM:29.3% propionic acid (PA)), leaving AA intact. Subsequent stages aimed to isolate AA by adsorbing the remaining low MW VFA (FW:58.9% BA, CKM:27.8% PA, 70% BA) other than AA, indicating effluent fractionation while preserving and purifying AA. Applied selective multi-stage adsorption approach offers a promising method to broaden waste-derived VFA applications.
Irritable bowel syndrome (IBS) is a common chronic gastrointestinal disorder, with diarrhea-predominant IBS (IBS-D) as the most frequent subtype. The implication of gut microbiota in the disease's etiology is not fully understood. In vitro gut systems can offer a great alternative to in vivo assays in preclinical studies, but no model reproducing IBS-related dysbiotic microbiota has been developed. Thanks to a large literature review, a new Mucosal ARtifical COLon (M-ARCOL) adapted to IBS-D physicochemical and nutritional conditions was set-up. To validate the model and further exploit its potential in a mechanistic study, in vitro fermentations were performed using bioreactors inoculated with stools from healthy individuals (n = 4) or IBS-D patients (n = 4), when the M-ARCOL was set-up under healthy or IBS-D conditions. Setting IBS-D parameters in M-ARCOL inoculated with IBS-D stools maintained the key microbial features associated to the disease in vivo, validating the new system. In particular, compared to the healthy control, the IBS-D model was characterized by a decreased bacterial diversity, together with a lower abundance of Rikenellaceae and Prevotellaceae, but a higher level of Proteobacteria and Akkermansiaceae. Of interest, applying IBS-D parameters to healthy stools was not sufficient to trigger IBS-D dysbiosis and applying healthy parameters to IBS-D stools was not enough to restore microbial balance. This validated IBS-D colonic model can be used as a robust in vitro platform for studies focusing on gut microbes in the absence of the host, as well as for testing food and microbiota-related interventions aimed at personalized restoration of gut microbiota eubiosis.
Anaerobic fermentation (AF) processes are sensitive to temperature fluctuations, which can influence the microbial activity and overall metabolic performances. Anaerobic reactors can face unforeseen temperature control failures, leading to instabilities in the process. The present study investigated the effect of two short-term temperature perturbations (down to 20°C and 15°C) on AF of food wastes (FWs). While 20°C did not exhibit a negative impact on AF performance maintaining the bioconversion yields over 40%, the reactor subjected to 15°C presented an acidogenic limitation, which decreased the bioconversion yields (36.4 ± 1.8%). As a result, 2.2 ± 0.5 g/L of succinic acid was accumulated in the reactor, being identified as a temperature failure indicator. Once the conditions were reestablished (operation temperature of 25ºC), the metabolic redundancies identified in the reactors allowed the AFs recovery to initial fermentation yields. 20°C was further tested as operational temperature resulting in stable bioconversion yield similar to the Control Reactor (43.2 ± 0.3%). These results showed the feasibility of conducting AF under low temperatures, indicating the potential of this technology to increase the cost-effectiveness of AF at psychrophilic conditions.
Polyhydroxyalkanoates (PHA) are bioplastics produced by few bacteria as intracellular lipid inclusions under excess carbon source and nutrient-deprived conditions. These polymers are biodegradable and resemble petroleum-based plastics. The rising environmental concerns have increased the demand for PHA, but the low yield in wild-type bacterial strains limits large-scale production. An improvement in the PHA production can be achieved by genetically engineering the wild-type bacterial strains by removing competitive pathways that divert the metabolites away from PHA biosynthesis, cloning strong promotors to overexpress the genes involved in PHA biosynthesis and constructing non-native metabolic pathways that feed the metabolites for PHA production. The desired monomers in the PHA polymers were obtained by elimination of genes involved in PHA biosynthetic pathway. The chain length degradation specific-gene deletion of β-oxidation pathway resulted in the accumulation of PHA monomers having high carbon chain length. A controlled accumulation of monomers in the PHA polymer was achieved by constructing novel pathways in the bacteria and deleting native genes of competitive pathways from the genome of non-PHA producers. The present review attempts to showcase the novel genetic modification approaches conducted so far to enhance the PHA production with a special focus on metabolic pathway gene deletion in various bacteria.
Gene editing is emerging as a powerful tool for introducing novel functionalities in mushrooms. While CRISPR/Cas9-induced double-strand breaks (DSBs) typically rely on non-homologous end joining (NHEJ) for gene disruption, precise insertion of heterologous DNA in mushrooms is less explored. Here, we evaluated the efficacy of inserting donor DNAs (8-1008 bp) with or without homologous arms at Cas9-gRNA RNP-induced DSBs. Co-transformation of donor DNAs with RNP targeting the pyrG gene in Ganoderma lucidum yielded 184 transformants without homologous arms and 781 with 300-bp homologous arms (HR_donor DNAs). Restriction analysis and sequencing identified 122 hR_donor DNA transformants with complete donor DNA sequences, achieving 15.6% HDR efficiency (122/781), contrasting with 8 instances via NHEJ from the 184 transformants. These findings highlight the viability of HDR for precise genomic editing in mushrooms, enabling targeted modifications to enhance functionalities.
This article presents new data on the integrated use of colloidal solutions of nanoparticles and low-intensity laser radiation on the biosynthetic activity of the medicinal mushroom Inonotus obliquus in vitro. Traditional mycological methods, colloidal solutions of biogenic metals, and unique photobiological methods have also been used. It was found that colloidal solutions of nanoparticles of all metals used increased the growth characteristics of I. obliquus (55-60%), while irradiation of the fungal inoculum with laser light in a medium with nanoparticles reduced the growth activity of I. obliquus mycelia by 12.3-35.4%. Silver nanoparticles (AgNPs) in a nutrient medium suppressed the biosynthesis of extracellular polysaccharides, whereas laser irradiation in the same medium increased the synthesis of intracellular polysaccharides by 9.7 times. Magnesium nanoparticles (MgNPs) and iron nanoparticles (FeNPs) inhibited the synthesis of intracellular polysaccharides in the mycelial mass of I. obliquus. At the same time, laser irradiation of the inoculum with MgNPs, on the contrary, induced a sharp increase in the amount of polysaccharides in the culture liquid (20 times). Treatment of the inoculum in a medium with nanoparticles with a laser caused an intensification of the synthesis of flavonoids in the mycelial mass and an increase in the synthesis of melanin pigments (25-140%). The results obtained suggest the possibility of the complex use of colloidal solutions of Fe, Ag, and Mg nanoparticles and low-intensity laser radiation as environmentally friendly factors for regulating biosynthetic activity in the biotechnology of cultivating the valuable medicinal mushroom I. obliquus.
Invasive fungal infections (IFIs) are responsible for elevated rates of morbidity and mortality, causing around of 1.5 million deaths annually worldwide. One of the main causative agents of IFIs is Candida albicans, and non-albicans Candida species have emerged as a spreading global public health concernment. Furthermore, COVID-19 has contributed to a boost in the incidence of IFIs, such as mucormycosis, in which Rhizopus oryzae is the most prevalent causative agent. The effector host immune response against IFIs depends on the activity of T cells, which are susceptible to the regulatory effects triggered by fungal virulence factors. The fungal cell wall plays a crucial role as a virulence factor, and its remodeling compromises the development of a specific T-cell response. The redirection of Jurkat T cells to target Candida spp. by recognizing targets expressed on the fungal cell wall can be facilitated using chimeric antigen receptor (CAR) technology. This study generated an M-CAR that contains an scFv with specificity to α-1,6 mannose backbone of fungal mannan, and the expression of M-CAR on the surface of modified Jurkat cells triggered a strong activation against Candida albicans (hyphae form), Candida tropicalis (hyphae form), Candida parapsilosis (pseudohyphal form), and Candida glabrata (yeast form). Moreover, M-CAR Jurkat cells recognized Rhizopus oryzae spores, which induced high expression of cell activation markers. Thus, a novel Mannan-specific CAR enabled strong signal transduction in modified Jurkat cells in the presence of Candida spp. or R. oryzae.
In the dynamic realm of healthcare, the convergence of engineering and biomedical sciences has emerged as a pivotal frontier. In this review we go into specific areas of innovation, including medical imaging and diagnosis, developments in biomedical sensors, and drug delivery systems. Wearable biosensors, non-wearable biosensors, and biochips, which include gene chips, protein chips, and cell chips, are all included in the scope of the topic that pertains to biomedical sensors. Extensive research is conducted on drug delivery systems, spanning topics such as the integration of computer modeling, the optimization of drug formulations, and the design of delivery devices. Furthermore, the paper investigates intelligent drug delivery methods, which encompass stimuli-responsive systems such as temperature, redox, pH, light, enzyme, and magnetic responsive systems. In addition to that, the review goes into topics such as tissue engineering, regenerative medicine, biomedical robotics, automation, biomechanics, and the utilization of green biomaterials. The purpose of this analysis is to provide insights that will enhance continuing research and development efforts in engineering-driven biomedical breakthroughs, ultimately contributing to the improvement of healthcare. These insights will be provided by addressing difficulties and highlighting future prospects.
Oleuropein (OP) is an appreciated compound present not only in fruits but also in leaves of olive trees, which can be transformed into hydroxytyrosol (HT), a substance with high antioxidant activity. In this work, the transformation of an agricultural residue containing OP (olive leaves or wastewater from mills) to the high added value compound HT is accomplished through different enzymatic strategies. Different enzymes were used, immobilized on various supports by diverse binding forces: beta-glucosidase encapsulated in siliceous material, esterases and lipases immobilized on hydrophobic supports (octyl-functionalized amorphous silica and periodic mesoporous organosilica), and esterase immobilized on amine-functionalized ordered mesoporous silica. All these biocatalysts were tested for oleuropein hydrolysis through two different reaction approaches: a) split of glucosidic bond catalyzed by beta-glucosidase (β-glu), followed by hydrolysis of the aglycon and further ester hydrolysis. 5 mg·mL-1 of β-glu fully hydrolyzed 5 mM OP at pH 7 and 50°C in 7 days, and further enzymatic hydrolysis of the aglycon yielded near to 0.5 mM HT in the best conditions tested. b) via direct hydrolysis of the ester bond to produce hydroxytyrosol in a one-step reaction using esterases or lipases. The latter reaction pathway catalyzed by lipase from Penicillium camemberti immobilized on octyl-silica (4 mg·mL-1) at 35°C and pH 6 directly produced 6.8 mM HT (1 mg·mL-1), transforming in 12 days near to 30% of the initial 25 mM OP from a commercial olive leaves extract.
Fiberbanks refer to a type of fibrous sediment originated by the forestry and wood pulping industry in Sweden. These anthropogenic sediments are significantly contaminated with potentially toxic elements, and a diverse array of organic pollutants. Additionally, these sediments are of environmental concern due to their potential role in greenhouse gas emissions. Given the environmental risks posed by these sediments, the development of effective remediation strategies is of critical importance. However, no specialized methods have been established yet for the cleanup of this specific type of contaminated sediments. To identify effective fungal species for the mycoremediation of the fiberbank substrate, we performed a detailed screening experiment. In this research, we primarily aimed at assessing both the growth capacity and the proficiency in degrading organic pollutants of 26 native white-rot fungi (WRF) species. These species were sourced from natural forest environments in northern Sweden. The experimental setup involved evaluating the WRF on plates containing fiberbank material with a central Hagem-agar disc to closely monitor the interaction of these species with fiberbank substrates. Among the fungi tested, Laetiporus sulphureus exhibited the highest growth area percentage at 72%, followed by Hymenochaete tabacina at 68% and Diplomitoporus crustulinus at 67%. For the removal of 2-3 ring polycyclic aromatic hydrocarbons (PAHs), Phellinus punctatus led with 68%, with Cystostereum muraii at 57% and Diplomitoporus crustulinus at 49%. Regarding the removal percentage of 4-6 ring PAHs, Diplomitoporus crustulinus showed the highest efficiency at 44%, followed by Phlebia tremellosa at 40% and Phlebiopsis gigantea at 28%.
Enzymatic hydrolysis of organic waste is gaining relevance as a complementary technology to conventional biological treatments. Moreover, biorefineries are emerging as a sustainable scenario to integrate waste valorization and high-value bioproducts production. However, their application on municipal solid waste is still limited. This study systematically evaluates the techno-economic feasibility of the conversion of the organic fraction of municipal solid waste (OFMSW) into high-value bioproducts through enzymatic hydrolysis. Two key variables are examined: (a) the source of the enzymes: commercial or on-site produced using OFMSW, and (b) the treatment of the solid hydrolyzate fraction: solid-state fermentation (SSF) for the production of biopesticides or anaerobic digestion for the production of energy. As a result, four different biorefinery scenarios are generated and compared in terms of profitability. Results showed that the most profitable scenario was to produce enzymes on-site and valorize the solid fraction via SSF, with an internal rate of return of 13%. This scenario led to higher profit margins (74%) and a reduced payback time (6 years), in contrast with commercial enzymes that led to an unprofitable biorefinery. Also, the simultaneous production of higher-value bioproducts and energy reduced the economic dependence of OFMSW treatment on policy instruments while remaining energetically self-sufficient. The profitability of the biorefinery scenarios evaluated was heavily dependent on the enzyme price and the efficiency of the anaerobic digestion process, highlighting the importance of cost-efficient enzyme production alternatives and high-quality OFMSW. This paper contributes to understanding the potential role of enzymes in future OFMSW biorefineries and offers economical insights on different configurations.
