Frontiers in Bioengineering and Biotechnology最新文献

Introduction: Prolonged air leaks (PAL) are considered to be one of the leading causes of postoperative complications following lung surgery. There are currently no clinically relevant methods for efficiently and systematically evaluating the underlying causes of PAL. Here, we introduce a new intuitive, physiologically-representative system for ex vivo negative pressure ventilation of lungs, equipped with PAL-oriented features.

Methods: Reproducibility and system capabilities were assessed using a lung simulation model capable of controlling the effective area of the defect, then validated with ex vivo specimens.

Results: Our system enables dynamic control of ventilation using either pressure (PCV) or volume (VCV) targets, with respective standard deviations of ±0.08 cm H2O and ±2.1 mL with moderate air leaks (<1,000 mL/min) and respective standard deviations of ±0.18 cm H2O and ±11 mL with severe air leaks (>1,000 mL/min). Additionally, leak quantification features proved comparable to that of the Thopaz+ (Medela Healthcare, Baar, Switzerland), a standard commercial digital thoracic drainage system, offering sufficient resolution to differentiate among clinically relevant air leaks. In the lower leak ranges (<400 mL/min) across all methods of evaluations, there were no significant differences between measured leak rates. For higher leak ranges, although there remained no significant differences between the Ex-PALM methods evaluated, the Thopaz + proved to systematically report lower leak rates values (Thopaz+ 420.0 ± 10.0 mL/min vs. PCV-derived 449.0 ± 19.9 mL/min, p < 0.05) and (Thopaz+ 1,200.0 + 0.0 mL/min vs. PCV-derived 1,239.7 ± 21.1 mL/min, p < 0.001). Unlike current systems, coughing was predictably replicated using peak pressure targets ranging from 100 to 300 cm H2O with a standard deviation of ±1.30 cm H2O from target. Our system allows extraction of biomechanical parameters at every breath, with theoretically expected pressures matching experimental measurements with a goodness fit value (R2) above 0.95 for the vast majority of breaths.

Discussion: The Ex vivo Pulmonary Air Leak Model (Ex-PALM) provides a preclinical PAL testing platform with high translational potential and applications in studying biomechanical mechanisms of PAL and developing intraoperative mitigation strategies.

With the synergistic advancement of micro/nanotechnology and intelligent control systems, medical micromachines are emerging as promising alternatives to conventional diagnostic and therapeutic methods, offering enhanced operational precision and minimal invasiveness for precision medicine applications. However, most existing micromachines rely on artificial synthetic materials, which involve complex micro-nano fabrication and raise biosafety concerns regarding immunogenicity and limited long-term therapeutic efficacy in deep tissues. The integration of natural biological cells with programmable optical tweezer has opened new avenues to overcome these limitations, enabling precise behavioral regulation and in situ assembly of cell-based micromachines. This review systematically outlines the design strategies underlying five categories of light force-powered cellular micromachines, including chemotactic bacteria, photosynthetic microalgae, red blood cells (RBCs), immune cells and subcellular structures, and highlights their pioneering applications in targeted drug delivery, minimally invasive surgery and desired immunotherapy. Meanwhile, it also addresses key challenges such as limited tissue penetration depth, phototoxicity management and operation intelligence, while suggesting future directions like adaptive optics-driven swarm control, optomechanobiological coupling and bioprinting-integrated systems. Additionally, the convergence of photonic technology, synthetic biology and artificial intelligence is expected to advance these micromachines into next-generation biomedical platforms for health supervision and disease therapy in vivo.

[This corrects the article DOI: 10.3389/fbioe.2022.945149.].

Gene and genome editing therapies are increasingly connected with nanomaterials, which protect and transport fragile nucleic acids and CRISPR/Cas systems through biological barriers safely and accurately. This review discusses how different nanocarriers, including lipid-based, polymeric, inorganic, and vesicle-derived systems, can improve delivery efficiency, cell targeting, endosomal escape, and intracellular movement for gene and genome editing. It summarizes findings from early clinical and preclinical studies, comparing several carrier types such as ionizable lipid nanoparticles, polymeric nanoparticles, micelles, gold and silica nanostructures, and engineered extracellular vesicles. The review also explains how specific design factors, such as surface ligands, charge modification, PEGylation, and stimulus-responsive behaviors, influence biodistribution, and improve on-target efficiency while lowering immune responses and off-target effects. Ethical and regulatory concerns for in vivo editing are highlighted, along with current methods used to study nano-bio interactions. Among these carriers, ionizable lipid nanoparticles show the most advanced performance for delivering nucleic acids and CRISPR systems. However, new polymer-based and exosome-inspired carriers are progressing rapidly for repeated and targeted applications. Hybrid and responsive systems may also enable better spatial and temporal control of editing. Future research should focus on stronger in vivo potency testing, improved biocompatibility evaluation, and standardized manufacturing to ensure clinical safety and reliability.

Glioblastoma GBM: Glioblastoma multiforme (GBM) remains one of the most lethal and treatment-resistant brain cancers, driven in part by the complexity of its tumour microenvironment (TME). While organ-on-chip (OoC) platforms offer more physiologically relevant models than traditional 2D or static 3D systems, their design remains largely empirical, lacking predictive control over flow conditions, biochemical gradients, and mechanical cues. Computational Fluid Dynamics (CFD) has emerged as a powerful tool to enhance the design, precision, and biological fidelity of OoC platforms. This comprehensive review highlights current limitations in replicating GBM's biological complexity and technical constraints in device fabrication and maintenance, mapping them to specific CFD strategies. It synthesises current strategies into a structured workflow for integrating CFD into the design, optimisation, and validation of microfluidic tumour models-bridging engineering precision with biological complexity. In addition, validation frameworks reported in the literature are highlighted and mapped onto GBM-on-chip applications have been recommended, drawing on widely recognised international standards for engineering validation and regulatory modelling practices. Finally, this review positions CFD as a core component of GBM-on-chip development and explores how its integration with AI-based optimisation can advance the creation of more predictive, scalable, and biologically relevant in vitro tumour models.

Background: Iron overload-related osteoporosis has garnered significant attention, yet its pathological mechanisms remain unclear. Traditional two-dimensional (2D) culture systems often fail to recapitulate the extracellular matrix (ECM) microenvironment, leading to discrepancies between in vitro and in vivo findings.

Methods: We developed a three-dimensional (3D) culture system using methacrylated gelatin (GelMA) microspheres to culture preosteoblastic cells, simulate the bone microenvironment under iron overload conditions, and systematically examine changes in cellular morphology, viability, function, and gene expression.

Results: Iron overload impaired cell viability, induced oxidative stress, and inhibited osteogenesis in both 2D and 3D cultures. However, cells in 3D exhibited enhanced resilience, including reduced ROS levels, higher viability, preserved cytoskeletal integrity, and less apoptosis and G1-phase arrest. Compared to 2D, 3D-cultured cells showed downregulated expression of ITGA1 and ITGB1, decreased adhesion function, and promoted proliferation. Transcriptomics further revealed activation of NF-κB signaling and DNA replication pathways in 3D, while key pathways such as Hippo, focal adhesion, and Wnt were suppressed.

Discussion: The GelMA microsphere-based 3D system provides a physiologically relevant model for studying iron overload. These findings offer not only mechanistic insights but also suggest potential microenvironment-targeted therapeutic strategies for iron overload-associated osteoporosis.

Introduction: Urinary catheterization frequently causes urinary tract infections and patient discomfort. While hydrogel coatings combining antibacterial and hydrophilic properties offer a potential solution, challenges such as uncontrolled antimicrobial release and poor coating adhesion limit their clinical utility. This study aimed to develop a novel dual-function hydrogel coating with controlled antibacterial activity and sustained lubrication for infection-resistant urinary catheters.

Methods: A bilayer PL@SAMT/Mg coating consisting of an inner polydopamine layer loaded with the antimicrobial peptide LL-37 and an outer pH-responsive MgO@AAm/SA/TA hydrogel was fabricated via surface modification and UV crosslinking. The coating was applied to catheters via surface modification followed by UV-induced crosslinking. It was characterized using SEM, EDS, FTIR, rheometry, and friction tests. Its antibacterial efficacy was evaluated against Staphylococcus aureus and Escherichia coli at different pH levels. Cytocompatibility was assessed using CCK-8, live/dead staining, and ELISA assays with L929, SV-HUC-1, and RAW264.7 cells. In vivo biocompatibility and antibacterial performance were investigated using a rat subcutaneous implantation model.

Results: The PL@SAMT/Mg coating exhibited a uniform, adherent bilayer structure with stable mechanical properties. It delivered excellent hydration lubrication and demonstrated pH-responsive swelling behavior. The release of LL-37 was sustained, while MgO release was significantly accelerated under alkaline conditions mimicking infection. The coating showed strong, pH-enhanced antibacterial activity against both S. aureus and E. coli. In vitro assays confirmed excellent cytocompatibility, anti-inflammatory effects, and anti-adhesion properties. In vivo, the coating minimized inflammation and significantly reduced bacterial colonization compared to uncoated catheters.

Discussion: The PL@SAMT/Mg coating successfully integrates intelligent antibacterial function with sustained lubrication. The PDA layer enables long-term preventive release of AMPs, while the pH-responsive hydrogel layer provides on-demand MgO release during infection. This synergistic, controlled-release strategy mitigates biotoxicity and resistance risks. The coating addresses key limitations of existing technologies through robust adhesion, effective antibacterial action, and biocompatibility, offering a promising approach to improve catheter performance and patient comfort.

Understanding pediatric phonation requires models that capture the biomechanical properties and dynamic airflow interactions of vocal folds. While synthetic vocal fold models have advanced the study of airflow-structure interactions in phonation, they cannot incorporate biologically relevant components such as hydrogels or human-derived cells. We developed a hybrid vocal fold phonatory platform that integrates a natural hydrogel with a silicone-based synthetic framework to address this limitation, enabling biomechanical fidelity and biological relevance. We adapted and downscaled a human vocal fold model (EPI) to replicate the dimensions of infant vocal folds. Using silicone elastomers and gelatin-silicone composites, we fabricated infant-scale replicas that mimic native tissue. Our results demonstrate that the material properties and geometrical scaling significantly affect vibratory behavior and acoustic output. Size reduction aligns with pediatric anatomical dimensions and minimizes the cell volume required for future biologically active models. This platform offers a scalable and bio-integrative approach for studying pediatric phonation, with potential applications in voice biomechanics, developmental vocal fold pathology, and tissue engineering.

Efficient delivery of biological material to the central nervous system remains a key limitation of conventional gene therapies. Recently, we developed a novel strategy based on a secretable and cell-penetrating TATk-CDKL5 fused protein which enhances the brain biodistribution and the therapeutic efficiency of the gene therapy approach in a mouse model of CDKL5 Deficiency Disorder (CDD). Here, to compare the efficacy of the TATk-CDKL5 gene therapy with a conventional approach in correcting the CDKL5 Deficiency Disorder pathological phenotype, we employed cortical organoids generated from CDD patient-derived iPSCs as a human model of CDD. We found greater therapeutic efficacy of the recombinant TATk-CDKL5 protein compared to the CDKL5 protein alone in improving or ameliorating defects caused by the absence of CDKL5, such as abnormal hyperexcitability evaluated with microelectrode arrays (MEA). Interestingly, CDD cortical organoids exhibited reduced cell proliferation and increased neuronal cell death compared to control cortical organoids; defects that were only restored by the expression of the recombinant TATk-CDKL5 protein. Based on the results from phenotypic and functional readouts, these findings suggest that gene therapy using a cross-correction approach offers superior efficiency in treating CDD.

The meniscus plays a crucial role in knee joint function, yet its limited intrinsic regenerative capacity, particularly within the avascular region, makes meniscal injuries a major therapeutic challenge. Meniscus progenitor cells (MPCs) have shown potential to promote meniscal regeneration, but their efficacy may be compromised in the inflammatory microenvironment. Joint lavage, a simple and clinically applied procedure that reduces intra-articular inflammation, may enhance the outcomes of cell-based therapies. Here, we investigated the therapeutic efficacy of combining joint lavage with MPC transplantation in rat partial meniscectomy models and further examined the effects of interleukin-1β (IL-1β), a key inflammatory mediator, on MPC function in vitro. Lavage significantly reduced intra-articular levels of multiple inflammatory cytokines, including IL-1β, which otherwise impaired MPC migration, proliferation, chondrogenic differentiation, and gene expression. In vivo, combined lavage and MPC therapy promoted more complete meniscal regeneration, enhanced type II collagen deposition, preserved cartilage integrity, and improved biomechanical properties compared with either treatment alone. These findings demonstrate that preconditioning the joint microenvironment through lavage markedly augments the regenerative potential of MPCs, providing a simple, low-cost, and clinically feasible strategy to improve meniscus and cartilage repair under inflammatory conditions.