Biotechnology Journal最新文献

Skin wound repair, a highly integrated and overlapping complex process, is susceptible to infection, excessive inflammation, high oxidative stress, which can prolong the wound healing time or even lead to chronic wound. Meanwhile, single-layer hydrogels are unavoidably dehydrated usually due to water evaporation, which is not conducive to wound healing. Therefore, a bilayer hydrogel dressing composed of gelatin (G) and sodium alginate (A) loaded with ciprofloxacin (C) and curcumin (Cur) was manufactured by extrusion 3D bioprinting technology in this study, which is denoted as GACCur. The experimental results showed that the bilayer hydrogel loaded with 50 µg/mL of Cur (GACCur50) had excellent antibacterial properties and high antioxidative activity with a 54.35 ± 0.48% DPPH scavenging ratio. In addition, this GACCur50 bilayer hydrogel exhibited the capacity to mitigate the polarization of inflammatory cells toward pro-inflammatory states, observably reducing the expression of pertinent pro-inflammatory cytokines. Furthermore, the bilayer hydrogel dressing also showed good biocompatibility as assessed by live-dead fluorescent staining, scanning electron microscopy (SEM), and CCK-8 test. Therefore, the GACCur50 bilayer hydrogel dressing is promising for wound healing.

In this work, double-layer heterogeneous CA scaffolds were designed for alveolar bone defects. The outer layer featured high hardness and slow degradation, and large pores and rapid degradation characterized the inner layer. The CA scaffold morphology was akin to bone defects, and its direct implantation reduced the operation time. A higher concentration of CA resulted in smaller pores and slower degradation. CA can promote the formation of mineralized nodules and the expression of genes related to mineralization without inducing cytotoxic effects. It also promoted the expression of cellular inflammatory factors, potentially through the TLR4 pathway. In vivo studies confirmed that CA did not promote the aggregation of inflammatory cells or the expression of inflammatory factors. In conclusion, the scaffold's characteristics of high surface hardness and slow degradation were beneficial for surface osteogenesis and maintaining the defect's shape and osteogenic space. Conversely, rapid internal degradation favors the formation of bone tissue.

Rheumatoid arthritis (RA) is a chronic autoimmune disease that affects synovial joints, leading to joint destruction, impaired physical function, and reduced quality of life. However, no accurate method for assessing RA risk currently exists. Given the critical role of early detection and intervention in RA management, further comprehensive risk assessments are essential. Mitochondrial oxidative stress (MOS) is a key factor in the initiation and progression of RA. The bidirectional interaction between RA and MOS supports the feasibility of MOS-based risk stratification for RA. Using public databases, we first applied the weighted gene co-expression network analysis (WGCNA) model to identify key genes involved in RA among MOS-related genes. Differential expression patterns of MOS-related genes were then analyzed using various machine learning algorithms to identify potential biomarkers. A nomogram model was established using CDKN1A, GADD45B, and MAFF genes to predict RA risk, followed by an evaluation of its reliability and stability. Additionally, we analyzed MOS-associated molecular subtypes and immune infiltration characteristics. Our findings highlight the significant role of MOS in RA development and underscore the clinical value of personalized treatment strategies.

This study conceptualizes the design of a small-scale system (250 mL–1 L) for the autotrophic cultivation of hydrogen-oxidizing bacteria, such as the representative strain Cupriavidus necator. The research aimed to systematically investigate the impact of bottle volume and gas composition, particularly oxygen concentration, on the growth and performance of C. necator during autotrophic cultivations. To this end, customized, pressure-tight, baffled glass bottles of various sizes (250, 500, and 1000 mL) and gas mixtures with varying oxygen concentrations (4%, 8%, and 12% v/v) were tested. Growth was monitored by measuring optical density. The maximum specific growth rate (µ_max), the biomass production rate (BPR), the volumetric gas–liquid mass transfer coefficient (k_La), and the oxygen transfer rate were calculated. Among the various combinations, the 1000-mL bottles demonstrated the highest µ_max (0.13 h⁻¹) and the second-highest BPR (0.074 g L⁻¹ h⁻¹) at an oxygen concentration of 8%, without the need to refill the headspace. The proposed small-scale system offers a swift and replicable method for concurrently investigating multiple autotrophic cultivations. In this regard, increasing the size of the bottle flask proved to be an efficient strategy to minimize the periodicity for gas refilling. Due to the inhibitory effect of oxygen, changing the liquid–gas volume ratio in hydrogen-driven shake flask cultivation had so far strongly influenced the growth rate. Our results provide a solid foundation for the scaling and optimization of small-scale cultivation of chemolithotrophic bacteria and will facilitate future parallelization and, hence, optimization of metabolic aspects.

Extracellular vesicles (EVs) are nanosized vesicles released by cells, containing molecular cargo such as proteins and nucleic acids. EVs offer promising avenues for the detection of biomarkers of disease and are excellent candidates for drug delivery and therapeutics. Although EVs can be obtained from biological fluids, it is challenging to obtain intact EVs from complex fluids and there is no universally accepted standard method of isolating EVs. When affinity chromatography-based isolation is used to isolate EVs from complex biofluids, there exist multiple ways to elute intact EVs from capture. This review aims to identify effective EV elution methods for preserving EV integrity and bioactivity after capture on aptamer-functionalized substrates, addressing the requirements of various downstream applications. We hypothesize that when used for elution, different materials and techniques influence the characteristics of EVs, such as their molecular content and bioactivity. The elution reagent and technique must be selected for the intended application for isolated EVs. However, currently, there is no agreement on the optimal elution method for EVs. This literature review aims to evaluate the different methods used to elute intact EVs from capture with regards to the downstream applications of isolated EVs. Based on the results of our analysis of recent literatures, the two elution reagents that are optimal for general purposes of the eluted intact EVs are deoxyribonuclease I and complementary oligonucleotides, as they both preserve EV characteristics that are required for molecular analysis and bioactivity, such as maintained morphology and protein profiles.

Triple-negative breast cancer (TNBC) is a clinically aggressive subtype of breast cancer that remains an unmet medical need. Because TNBC cells do not express the most common markers of breast cancers, there is an active search for novel molecular targets in triple-negative tumors. Additionally, this subtype of breast cancer presents strong immunogenic characteristics which have been encouraging the development of immunotherapeutic approaches against the disease. In this context, nucleolin arises as a promising target for immunotherapy against TNBC. Our group has previously developed an anti-nucleolin VHH-Fc antibody capable of eliciting antibody-dependent cellular cytotoxicity (ADCC). Moreover, we constructed and characterized an antibody library, that was screened against nucleolin-overexpressing cells, originating an enriched anti-nucleolin antibody pool. In this work, a strategy to select individual clones from the pool was designed, combining NGS data with 3D modeling. Two antibodies demonstrated a significant 4.4- and 6.1-fold increase in binding to nucleolin-overexpressing and TNBC cells, and an improvement in affinity to the sub-micromolar range (0.19 µM and 83.69 nM). Additionally, an increment in 4.6- and 3.1-fold in ADCC activity against respective cell lines was observed for the M2 antibody clone. Herein, the affinity maturation strategy developed was validated and corroborated a positive, but not proportional, correlation between antibody binding, affinity, and ADCC.

Osteochondral damage, caused by trauma, tumors, or degenerative diseases, presents a major challenge due to the limited self-repair capacity of the tissue. Traditional treatments often result in significant trauma and unpredictable outcomes. Recent advances in bone/cartilage tissue engineering, particularly in scaffold materials and fabrication technologies, offer promising solutions for osteochondral regeneration. This review highlights the selection and design of scaffolds using natural and synthetic materials such as collagen, chitosan (Cs), and polylactic acid (PLA), alongside inorganic components like bioactive glass and nano-hydroxyapatite (nHAp). Key fabrication techniques—freeze-drying, electrospinning, and 3D printing—have improved scaffold porosity and mechanical properties. Special focus is placed on the design of multiphasic scaffolds that mimic natural tissue structures, promoting cell adhesion and differentiation and supporting the regeneration of cartilage and subchondral bone. In addition, the current obstacles and future directions for regenerating damaged osteochondral tissues will be discussed.

Due to their superior physicochemical features, chitosan thermosensitive hydrogels are multipurpose platforms that are frequently used in the biomedical industry. Many investigations have been conducted recently to modify their pore dimensions, expansion, biodegradability, stimulus-reaction characteristics, and other characteristics in order to better tailor them to the complex craniofacial tissues. They have been the focus of various studies that have attempted to load biological cargos for therapeutic and regenerative uses in the oro-facial tissues. This article assesses the utility and advancements of chitosan thermosensitive hydrogel-based pharmaceutical delivery devices for the management of craniofacial disorders such as dental caries, endodontic diseases, periodontitis, temporomandibular disorders, mucosal diseases, cancer, and so forth.