Bioengineered最新文献

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.

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.

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.

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.

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.

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.

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.