Frontiers in Bioengineering and Biotechnology最新文献

The keto acids of isoleucine and leucine are bioavailable precursors of their branched-chain amino acids in Chinese hamster ovary (CHO) cells, which are used to produce biotherapeutics at industrial scale. In this study, the potential of branched-chain keto acids to improve product yield was evaluated in fed-batch and simulated steady-state perfusion. In fed-batch, combined or individual replacement of isoleucine and leucine at equimolar concentrations with their keto acids moderately increased (+6%) or maintained the cell-specific productivity qP, but this positive impact was counteracted by a reduction in cell growth up to -11%. Higher concentrations of keto acids substantially reduced cell growth (-42%) and qP (-25%). ¹³C-metabolic flux analysis during the growth phase of the fed-batch revealed that this detrimental effect may be associated with impaired glycolysis and TCA cycle activity, along with altered fluxes in anaplerotic reactions, ultimately leading to decreased ATP (-20%) and NADPH (-29%) generation. In steady-state perfusion, keto acid supplementation improved IgG yield up to 100% through (I) reduced bleed rates as a result of lower cell growth and (II) enhanced qP. Untargeted metabolite profiling demonstrated altered levels of various compounds, suggesting pathways that may be associated with the observed improvements. Overall, the findings of this study highlight the potential of novel media components, such as keto isoleucine and keto leucine, to improve yields and efficiency in biopharmaceutical production, thereby contributing to increased sustainability and lower manufacturing costs.

Chronic diabetic wounds exhibit persistent oxidative stress, prolonged inflammation, impaired angiogenesis, and a disrupted bioelectric microenvironment that hinders re-epithelialization. Here, we develop an injectable Prussian Blue nanofiber-PVA hydrogel (PBM.PVA gel) with electroconductive and immunomodulatory features for accelerated diabetic wound repair. Electrospun PBM nanofibers were uniformly embedded within a physically cross linked PVA matrix, producing a flexible and adhesive composite with stable conductivity. In vitro, PBM. PVA gel showed excellent cytocompatibility, reduced pro-inflammatory cytokines (IL-6, TNF-α, CD86), and enhanced pro-regenerative markers (CD206, CD31). In streptozotocin-induced diabetic mice, the hydrogel significantly accelerated wound closure, reduced inflammatory infiltration, and promoted collagen deposition with increased CD31-positive staining. While PBM has been reported to possess redox-regulatory potential, ROS levels and endogenous wound electrical fields were not directly quantified in this study; therefore, mechanistic interpretations are described as plausible and require further validation. Together, PBM. PVA gel provides a multifunctional dressing that supports a favorable wound microenvironment and improves healing outcomes in diabetic wounds.

Cold atmospheric plasma (CAP) is a non-thermal plasma generated near room temperature that has broad medical applications in the medical field, including antitumor, antimicrobial, and anti-inflammatory effects, promotion of tissue regeneration, and enhancement of transdermal and mucosal drug delivery. However, there is currently a lack of standardization regarding the indications for CAP and its application parameters, resulting in varying degrees of histological penetration depths reported in different studies. Therefore, to further promote the safe and effective clinical application of CAP, the histological levels at which CAP can be applied must be clearly defined. Here, we review the depth of tissue penetration achieved by CAP under various conditions and analyze the key factors influencing penetration depth, using this knowledge to propose how these factors should be adjusted for different application requirements to achieve safer and more precise therapies.

Platelet-rich plasma (PRP) therapy, an autologous biologic rich in growth factors, has emerged as a promising modality in regenerative medicine, with expanding relevance in nursing-led care. PRP promotes tissue regeneration, modulates inflammation, and enhances functional recovery in several conditions, including chronic wounds, musculoskeletal disorders, and aesthetic applications. Its minimally invasive nature and patient-specific approach align closely with holistic nursing models. This review synthesizes current evidence on the biological mechanisms underlying PRP activity, including platelet composition, growth factor signaling, and immunomodulation, and critically evaluates clinical outcomes relevant to nursing practice. Particular attention is given to nursing roles throughout the PRP treatment continuum, from patient selection and preparation to monitoring and long-term follow-up. Despite its clinical promise, PRP adoption in nursing practice faces barriers such as formulation variability, lack of standardized protocols, and limited nurse-led guidelines. We propose strategic directions for integrating PRP into evidence-based nursing frameworks, emphasizing digital tools, individualized care pathways, and interdisciplinary collaboration.

Plant-derived extracellular vesicles (PDEVs) and plant-derived nanoparticles (PDNPs) are emerging plant-based nanomaterials with growing relevance in biotechnology, agriculture, and health. Although often grouped together, they arise from distinct origins: PDEVs are actively secreted vesicles with selective cargo loading, whereas PDNPs form during tissue disruption and reflect the lipid-metabolite composition of plant biomass. This review summarizes recent progress in distinguishing these systems, including advances in biogenesis, isolation, biomarkers, and functional characterization. We highlight mechanistic insights into PDEV-mediated cross-kingdom RNA communication in plant immunity and the strong translational potential of PDNPs in oral drug delivery, immunomodulation, and microbiome regulation. Remaining challenges include standardization, scalable purification, and deeper mechanistic clarity. By clarifying their differences and complementary strengths, this review outlines a foundation for developing reliable plant-derived nanovesicle technologies.

Introduction: Adequate bone volume and contour are essential for successful implant placement. This study evaluated the accuracy of a novel self-positioning three-dimensional printed individualized titanium mesh (3D-PITM) in guided bone regeneration (GBR).

Methods: Ten identical maxillary phantoms with standardized defects were divided into an experimental self-positioning 3D-PITM group and a conventional 3D-PITM group. Pre- and postoperative CBCT scans were obtained for 3D reconstruction and superimposition. Deviations in augmented contours, screw placement, volumetric accuracy, and 2D cross-sectional augmentation were analyzed.

Results: The self-positioning group showed significantly reduced deviation in augmentation contours (0.82 ± 0.07 mm vs. 1.02 ± 0.13 mm, P = 0.003), improved screw placement accuracy (0.10 ± 0.13 mm vs. 0.65 ± 0.32 mm, P = 0.026), and lower volumetric discrepancies. Two-dimensional evaluation confirmed greater vertical and horizontal accuracy in bone augmentation (P = 0.021, P = 0.018).

Conclusion: The self-positioning 3D-PITM achieved more accurate installation and predictable bone augmentation in vitro, suggesting potential clinical advantages for implant-supported rehabilitation.

Purpose: Anastomotic leakage and impaired healing remain major complications in gastrointestinal (GI) surgery. Small intestinal submucosa (SIS), a biological scaffold, has shown regenerative potential but its mechanisms in GI anastomotic healing remain unclear. This study aimed to investigate the effects of an SIS bio-patch on intestinal anastomotic healing, focusing on immune modulation, microbiota reshaping, and metabolic changes.

Methods: C57BL/6 mice underwent GI anastomosis with or without SIS bio-patch implantation. Five days post-operation, tissues were collected for histology, immunofluorescence, flow cytometry, 16S and ITS sequencing, and untargeted metabolomics. Immune cell composition, barrier protein expression, microbiota composition, and metabolic signatures were analyzed.

Results: SIS bio-patch significantly reduced inflammation and enhanced mucosal barrier integrity, as evidenced by reduced TNF-α and IL-6 and increased ZO-1 and occludin expression. SIS increased IL-22⁺ILC3s (type 3 innate lymphoid cells) and decreased the Th17/Treg ratio without altering macrophage polarization. Microbiota analysis showed increased abundance of Bifidobacterium and Alloprevotella, correlating positively with IL-22⁺ILC3s. Fungal sequencing revealed higher Fungi gen. Incertae sedis levels, associated with beneficial immune profiles. Metabolomics showed elevated amino acids and biotin metabolism in SIS-treated tissues, which may support epithelial regeneration.

Conclusion: SIS bio-patch promotes anastomotic healing by enhancing IL-22⁺ILC3-mediated repair, rebalancing adaptive immunity, reshaping microbial communities, and upregulating pro-regenerative metabolic pathways. These findings support the use of SIS as an immunomodulatory biomaterial for gastrointestinal repair.

Background: Squat training enhances athletic performance but poses knee injury risks when the technique is poor.

Objective: Develop a resistance-type wearable knee exoskeleton to cultivate a hip-dominant, knee-safe squat pattern.

Methods: Fifteen healthy men performed squats with either the exoskeleton or with barbells at three matched loads. Three-dimensional motion, ground-reaction force, and electromyography data were processed in OpenSim and MATLAB to quantify joint kinematics, power share, and muscle contribution.

Results: The exoskeleton significantly reduced average angular velocity at the hip, knee, and ankle (p < 0.05), increasing hip power contribution by 20%-40% while decreasing knee contribution by 20%-30%, confirming a hip-driven pattern. However, knee and ankle ranges of motion decreased by 7°-9°, and vastus medialis activation dropped by ∼50% (p < 0.05).

Conclusion: The device effectively standardizes squat mechanics and off-loads the knee, yet individualized tuning and auxiliary mobility work are recommended to optimize training transfer and preserve functional range of motion.

Background: Endothelial dysfunction is increasingly recognized as an early and central event in the onset of cardiovascular and neurodegenerative diseases. Ginkgo biloba extracts are known for their vascular-protective properties, including enhancement of endothelial function, antioxidant and anti-inflammatory activity, nitric oxide preservation, and modulation of platelet aggregation. Plant-derived nanovesicles (PDNVs) are emerging as versatile bioactive carriers with demonstrated anti-inflammatory, anticancer, antimicrobial, regenerative, and microbiota-modulating effects. However, their vascular-protective potential remains underexplored. This study aimed to assess the effects of PDNVs isolated from Ginkgo biloba seeds on endothelial responses under inflammatory stress.

Methods: PDNVs were isolated from Ginkgo biloba seed homogenate using differential ultracentrifugation followed by density gradient ultracentrifugation with linear and non-linear iodixanol gradients. Nanoparticle tracking analysis (NTA) and cryo-transmission electron microscopy (cryo-TEM) characterized vesicle size, concentration, and morphology. Untargeted mass spectrometry profiled the protein content of distinct PDNV fractions. Functional assays were conducted on human umbilical vein endothelial cells (HUVECs) exposed to lipopolysaccharide (LPS)-induced inflammatory stress.

Results: Ginkgo PDNV isolates comprise a heterogeneous population of nanometer sized particles, including vesicles with single and double layers. Proteomics revealed seed storage proteins (legumin and ginnacin) and membrane-associated ATPases, HSP90, catalase, phosphoenolpyruvate carboxylase (PEPC), and eEF1A. PDNVs were non-toxic up to 50 μg/mL; at 100 μg/mL, they enhanced mitochondrial activity but triggered early apoptosis and necrosis. PDNVs did not increase ROS production, even in the presence of H₂O₂. At 1 μg/mL, they significantly suppressed LPS-induced expression of IL-1β, TNF-α, IL-6, and IL-8 (mRNA and protein; p ≤ 0.05 to p ≤ 0.001). PDNVs preserved endothelial integrity by downregulating VCAM-1 and upregulating occludin, maintained eNOS expression (p ≤ 0.01), and attenuated COX-1, COX-2, and prostacyclin synthase (PGIS) induction. Thrombotic markers (TXB₂, vWF, and PAI-1) remained unaffected.

Conclusion: Ginkgo seed-derived PDNVs exhibit vascular-protective and anti-inflammatory properties, supporting their potential as safe, multifunctional agents for endothelial modulation. Further studies are warranted to explore their therapeutic applications in vascular biology.

Background: Aortoesophageal fistula (AEF) is a relatively rare and life-threatening condition, and the optimal surgical treatment for secondary AEF following thoracic endovascular aortic repair (post-TEVAR AEF) remains controversial. This study aimed to summarize the clinical efficacy of aortic arch debranching combined with extra-anatomic bypass for the treatment of post-TEVAR AEF.

Methods: The clinical data of 16 patients who underwent surgical treatment for post-TEVAR AEF at our institution from 30 June 2019 to 30 June 2024 were retrospectively reviewed. Aortic arch debranching and extra-anatomic aortic bypass under general anesthesia were performed for most patients. Empirical antibiotics were administered for 6-8 weeks. The acute and long-term outcomes were summarized.

Results: Stent-related infection leading to AEF occurred at a median interval of 30 months after the initial TEVAR surgery. All patients presented with recurrent fever preoperatively; blood bacterial cultures were positive in nine patients (56.25%) and negative in seven patients (43.75%). The median operative time was 460.5 (433.5, 543.5) minutes, and the median intensive care unit stay was 7 (5.25, 31.75) days. No intraoperative mortality was observed in this cohort. During the follow-up period, three patients developed recurrent AEF accompanied by severe infection. Four patients died postoperatively, including one who died of thoracic aortic rupture and hemorrhage within 3 months postoperatively, and three others died of multiple organ failure at 4-10 weeks after surgery. The remaining 12 patients achieved favorable postoperative recovery without the need for prolonged antibiotic therapy.

Conclusion: Aortic arch debranching and extra-anatomic bypass from the ascending aorta to the proximal abdominal aorta yields favorable acute and long-term outcomes for patients with post-TEVAR AEF.