Frontiers in Bioengineering and Biotechnology最新文献

The valorization of tobacco stems, a major agricultural by-product with high lignin content, is hindered by the recalcitrance of plant cell walls. Conventional approaches using single microbes or enzymes suffer from inefficiency, instability, and poor performance on lignin-rich substrates. To address this, we proposed a "microbe-enzyme synergistic" strategy by combining Bacillus megaterium (a multifunctional degrader) with its own extracellular enzymes. Three treatments were designed: microbe-only (T_b), enzyme-only (T_e), and microbe-enzyme combination (T_b&e), with sterile water as the control (CK). Cell wall component contents, enzyme activities, and microbial community structure were analyzed after 7 days of fermentation. The T_b&e treatment achieved the most substantial degradation: 64.93% for cellulose, 57.89% for lignin, and 37.20% for pectin, significantly outperforming T_b and T_e. It also exhibited the highest activities of cellulase, laccase, and pectinase. High-throughput sequencing revealed that T_b&e specifically enriched Bacillus megaterium (94.97% of the community) while suppressing non-functional competitors. Our findings demonstrate that the synergy creates a positive feedback loop: initial enzymatic hydrolysis facilitates microbial colonization, which in turn boosts sustained enzyme production. This study elucidates a mechanistic model for efficient biodegradation and provides an innovative strategy for the high-value utilization of high-lignin agricultural wastes.

Objective: Lumbar disc herniation (LDH) has demonstrated a rising prevalence in contemporary clinical practice, significantly compromising patients' daily lives and potentially necessitating surgical intervention. While annulus fibrosus suture (AFS) techniques are increasingly incorporated into surgical protocols, current evidence remains inconclusive regarding their definitive clinical advantages.

Methods: We performed a comprehensive search of eight databases from inception to September 2025 to identify published articles on AFS for LDH. Outcome measures encompassed operative time, incision length, blood loss, length of stay (LOS), visual analog scale (VAS) score, Japanese Orthopedic Association (JOA) score, Oswestry disability index (ODI) score, disc height, recurrence, and complication. The quality of the studies was analyzed using the RoB-2 and ROBINS-I tools. Statistical analyses were performed using RevMan 5.4 software and Stata 17 software. The GRADE approach was used to evaluate the certainty of evidence for outcomes.

Results: A total of 58 studies encompassing 5,765 patients diagnosed with LDH. The control group had shorter operative time (MD = 4.85, 95% CI: 2.79 to 6.92, P < 0.00001) compared to the AFS group. There were no differences in terms of incision length, blood loss, LOS, JOA score, ODI score, and complication between the techniques. AFS group demonstrated a significant benefit over control group in terms of VAS score (MD = -0.24, 95% CI: -0.33 to -0.14, P < 0.00001), disc height (SMD = 1.36, 95% CI: 0.73 to 2.00, P < 0.0001), and recurrence (RR = 0.34, 95% CI: 0.27 to 0.42, P < 0.00001). The results of subgroup analysis showed that different study types and different follow-up times were a source of heterogeneity. The quality of evidence for outcome measures ranges from very low to moderate.

Conclusion: Current evidence suggests that AFS therapy may be advantageous in improving clinical symptoms and may reduce postoperative recurrence. Due to limited data and low quality of evidence, additional large-scale, multicenter trials are needed to verify and strengthen these findings.

Sequence verification of plasmids is a fundamental process in synthetic biology. For plasmid sequence verification using next-generation sequencing (NGS) library preparation, Tn5 transposase is widely used. Streamlined sequencing workflow for laboratory-scale applications is important; however, recombinant Tn5 production in-house can be laborious. In this study, we demonstrated that the addition of a large soluble tag was not essential for purification and that the fusion of a His10 tag and protein A was sufficient to yield active Tn5 transposase in adequate amounts. In addition, we evaluated exonuclease-based genomic DNA digestion for plasmid sequencing from an E. coli lysate and the data analysis pipeline of sequences derived from the Illumina iSeq100 platform for de novo assembly, reference mapping, and annotation. This study proposes a simple workflow of an in-house easy-to-purify Tn5-based plasmid sequencing platform using a compact benchtop sequencer (AistSeq).

Introduction: Catheter-related infections and biofouling remain critical challenges in clinical practice due to limited surface functionalities and rapid bacterial biofilm formation.

Methods: We developed a universal bottom-up strategy to fabricate hierarchical inverse opal hydrogel coatings on medical catheters via dopamine-mediated substrate activation, dual-layer colloidal assembly, and polymer infiltration, followed by oil infusion to enable adaptive wettability and low-friction liquid mobility.

Results: The coatings exhibited stress-responsive wetting transitions, structural-color-enabled visual monitoring of degradation, and tunable droplet adhesion by modulating pore geometry. In vitro tests showed 98.9% antibacterial efficiency against E. coli, together with excellent hemocompatibility, cytocompatibility, and in vivo biosafety.

Discussion: By integrating passive antifouling, controlled drug release, and real-time structural feedback in a single interface, this platform provides a robust route toward infection-resistant and intelligent catheter devices.

Introduction: Synthetic polymers are widely used in scaffold fabrication but are generally bioinert and hydrophobic, limiting cell adhesion and interaction. Hyaluronic acid (HA) is a ubiquitously expressed polycarbohydrate and a key extracellular matrix component that regulates tissue hydration, cell adhesion, motility, and regeneration. Incorporating HA into synthetic materials presents a promising strategy to enhance hydrophilicity and support biological functions, such as cell adhesion and osteogenic differentiation. This study investigated the effects of HA coating on the physicochemical characteristics and osteogenic potential of poly(L-lactide-co-1,3-trimethylene carbonate) (PLATMC) scaffolds.

Methods: PLATMC scaffolds were coated with HA via immersion. HA stability during sterilization and culture was assessed alongside release kinetics. BMSC responses and osteogenic differentiation were evaluated in vitro and in vivo over six months. Scaffold wettability was analyzed to determine changes in surface hydrophilicity following HA coating.

Results: HA coating improved scaffold wettability in a concentration-dependent manner. HA release was characterized by burst kinetics, becoming undetectable after a few days under in vitro conditions, indicating that HA-driven effects are expected to be strongest during early cell-material interactions. In vitro, human BMSC showed RHAMM upregulation across all HA groups and CD44 upregulation in the 0.5% HA group after 24 hours. Rat BMSC exhibited increased osteocalcin expression, suggesting enhanced osteoinductive activity, corroborated by a trend toward osteopontin upregulation in human BMSC. In vivo, μCT analysis revealed higher tissue density surrounding HA-coated scaffolds at 8 weeks compared to 6 months in a rat subcutaneous implantation model.

Conclusion: HA coating improved scaffold hydrophilicity and promoted early cell adhesion and osteogenic signaling. The findings indicate that HA-coated PLATMC scaffolds support early cellular engagement and osteoconductivity, while long-term outcomes are likely governed by intrinsic scaffold properties. These results highlight the potential of HA-coated PLATMC scaffolds for biofabrication in dentistry, particularly in oral and maxillofacial bone regeneration.

This comprehensive review examines cutting-edge developments in hydrogel technology for thyroid disease management, encompassing both diagnostic and therapeutic applications. As a promising polymeric biomaterial with a three-dimensional (3D) network structure, hydrogels demonstrate exceptional potential in thyroid disease due to their unique combination of properties: (1) remarkable biocompatibility, (2) precisely tunable physicochemical characteristics, and (3) controlled drug release capabilities. Our analysis systematically evaluates hydrogel applications across the spectrum of thyroid disorders, including (i) diagnostic approaches for thyroid nodules, (ii) therapeutic interventions for endocrine dysfunction (hyperthyroidism, hypothyroidism, and post-thyroidectomy hypoparathyroidism), (iii) innovative treatments for thyroid neoplasms, and (iv) hemostasis, wound healing, repair of thyroid cartilage and laryngeal nerve injuries following thyroid surgery. It focuses on analyzing their advantages and challenges in drug delivery, minimally invasive therapy, tissue engineering, and postoperative care. Finally, future development directions for hydrogels in the field of thyroid disease are discussed.

Objective: To compare the clinical outcomes of locking plate versus cancellous screw fixation in structural autologous bone grafting for medial tibial bone defects during total knee arthroplasty.

Methods: A retrospective analysis was conducted on 66 patients with medial tibial bone defects who underwent TKA between January 2024 and December 2024. Among them, 34 patients received locking plate fixation (LP group), and 32 patients received cancellous screw fixation (CS group). Postoperative outcomes, including the hip-knee-ankle (HKA), Knee Society Score (KSS) and postoperative complications were recorded and compared to evaluate graft integration and functional recovery.

Results: All patients successfully completed the surgery. At the final follow-up, both groups showed significant improvement in HKA angle, KSS Knee Score, and KSS Function Score compared to preoperative values (P < 0.05). However, no statistically significant differences were observed between the two groups at any time point in terms of KSS Knee and Function Scores (P > 0.05). No postoperative complications occurred in either group during the follow-up period.

Conclusion: Compared with conventional cancellous screw fixation, the use of locking plate combined with structural autologous bone grafting provides similarly favorable clinical outcomes. Furthermore, locking plate offer broader intraoperative applicability and may serve as an effective internal fixation strategy for managing medial tibial bone defects during TKA.

Objective: Prototype foamy virus (PFV) is an attractive gene delivery platform due to its large cargo capacity and favorable safety profile; however, the structural basis of its interaction with heparan sulfate (HS), a critical attachment factor for viral entry, remains undefined. The objective of this study was to identify the structural determinants of HS recognition within PFV Env and to evaluate whether rational, structure-guided engineering could enhance viral entry and gene transfer efficiency.

Methods: We applied a structure-guided engineering strategy combining in silico structural modeling, molecular docking, and systematic mutagenesis of the PFV Env receptor-binding domain (RBD), targeted residue substitutions, and combinatorial mutations spanning the upper domain (UD) and lower domain (LD) were generated and evaluated using quantitative cell-based transduction assays. In addition, Tet-On-inducible Env-expressing stable producer cell lines were established to provide a reproducible platform for functional validation.

Results: Alanine substitutions at R298, R440, and E446 in the UD abolished infectivity, confirming their essential roles in HS-mediated attachment. In contrast, selective substitutions at adjacent positions, Q296R and G403F in the UD, and E232N, I330F, and I334F in the LD, enhanced transduction efficiency by up to 1.32-fold relative to the wild type. Combinatorial variants integrating beneficial UD and LD mutations exhibited synergistic effects, achieving a transduction efficiency of 68.9%, corresponding to a 1.55-fold increase over the wild type (44.4%). Interspecies domain replacement with simian foamy virus Env reduced infectivity, underscoring the context-specific nature of PFV-HS interactions. In the inducible stable cell system, the LD var6 mutant achieved 8.6% transduction compared to 4.4% for the wild type, representing up to a 1.95-fold increase.

Discussion: These findings define the structural determinants of HS recognition in PFV Env and demonstrate that residue-level, structure-guided engineering can enhance PFV transduction efficiency. This study provides experimentally validated insight into PFV Env-HS interactions and establishes a rational framework for further optimization of PFV-based gene delivery technologies.