Frontiers in Bioengineering and Biotechnology最新文献

Atherosclerotic cardiovascular disease (ACD) is the leading cause of death worldwide. The gold standard of treatment is the implantation of a permanent stent implant that is often associated with complications such as thrombus formation, vascular neointimal response, and stent fracture, which altogether decrease the long-term safety and efficacy of the stent. Biodegradable metallic materials have become an attractive alternative because of the ability to facilitate a more physiological healing response while the metal degrades. Recently, Molybdenum (Mo) has been considered as a potential candidate due to its excellent mechanical and medical imaging properties. Moreover, the biomedical research studies performed to date have shown minimal adverse effects in vitro and in vivo. However, there are still concerns of toxicity at high doses, and the impact of the biochemical mechanisms of Mo on material performance especially in pathophysiological environments are yet to be explored. Mo is an essential co factor for enzymes such as xanthine oxidoreductase (XOR) that plays a critical role in vascular homeostasis and ACD progression. Herein, this review will focus on the biochemistry of Mo, its physiological and pathological effects with an emphasis on cardiovascular disease as well as the recent studies on Mo for cardiovascular applications and its advantages over other biodegradable metals. The limitations of Mo research studies will also be discussed and concluded with an outlook to move this revolutionary metallic biomaterial from the bench to the bedside.

Introduction: 3D-printed scaffolds have emerged as an alternative for addressing the current limitations encountered in bone reconstruction. This study aimed to systematically review the feasibility of using 3D bio-printed scaffolds as a material for bone grafting in animal models, focusing on femoral and tibial defects. The primary objective of this study was to evaluate the efficacy, safety, and overall impact of these scaffolds on bone regeneration.

Methods: Electronic databases were searched using specific search terms from January 2013 to October 2023, and 37 relevant studies were finally included and reviewed. We documented the type of scaffold generated using the 3D printed techniques, detailing its characterization and rheological properties including porosity, compressive strength, shrinkage, elastic modulus, and other relevant factors. Before incorporating them into the meta-analysis, an additional inclusion criterion was applied where the regenerated bone area (BA), bone volume (BV), bone volume per total volume (BV/TV), trabecular thickness (Tb. Th.), trabecular number (Tb. N.), and trabecular separation (Tb. S.) were collected and analyzed statistically.

Results: 3D bio-printed ceramic-based composite scaffolds exhibited the highest capacity for bone tissue regeneration (BTR) regarding BV/TV of femoral and tibial defects of animal models. The ideal structure of the printed scaffolds displayed optimal results with a total porosity >50% with a pore size ranging between 300- and 400 µM. Moreover, integrating additional features and engineered macro-channels within these scaffolds notably enhanced BTR capacity, especially observed at extended time points.

Discussion: In conclusion, 3D-printed composite scaffolds have shown promise as an alternative for addressing bone defects.

Individuals with transfemoral lower limb amputations walk with adapted gait. These kinetic and kinematic compensatory strategies will manifest as differences in muscle recruitment patterns. It is important to characterize these differences to understand the reduced endurance, reduced functionality, and progression of co-morbidities in this population. This study aims to characterize muscle recruitment during gait of highly functional individuals with traumatic transfemoral amputations donning state-of-the-art prosthetics compared to able-bodied controls. Inverse dynamic and static optimisation methods of musculoskeletal modelling were used to quantify muscle forces of the residual and intact limb over a gait cycle for 11 individuals with traumatic transfemoral amputation and for 11 able-bodied controls. Estimates of peak muscle activation and impulse were calculated to assess contraction intensity and energy expenditure. The generalized estimation equation method was used to compare the maximum values of force, peak activation, and impulse of the major muscles. The force exhibited by the residual limb's iliacus, psoas major, adductor longus, tensor fasciae latae and pectineus is significantly higher than the forces in these muscles of the intact contralateral limb group and the able-bodied control group (p < 0.001). These muscles appear to be recruited for their flexor moment arm, indicative of the increased demand due to the loss of the plantar flexors. The major hip extensors are recruited to a lesser degree in the residual limb group compared to the intact limb group (p < 0.001). The plantar flexors of the intact limb appear to compensate for the amputated limb with significantly higher forces compared to the able-bodied controls (p = 0.01). Significant differences found in impulse and peak activation consisted of higher values for the limbs (residual and/or intact) of individuals with transfemoral lower limb amputations compared to the able-bodied controls, demonstrating an elevated cost of gait. This study highlights asymmetry in hip muscle recruitment between the residual and the intact limb of individuals with transfemoral lower limb amputations. Overall elevated impulse and peak activation in the limbs of individuals with transfemoral amputation, compared to able-bodied controls, may manifest in the reduced walking endurance of this population. This demand should be minimised in rehabilitation protocols.

The simultaneous administration of antibacterial treatment and acceleration of tissue regeneration are crucial for the effective healing of infected wounds. In this work, we developed a facile hydrogel (PCC hydrogel) through coordination and hydrogen interactions by polymerizing acrylamide monomers in the presence of carboxymethyl chitosan nanoparticles and copper ions. The prepared PCC hydrogel demonstrated effective bacterial capture from wound exudation and exhibited a potent bactericidal activity against methicillin-resistant Staphylococcus aureus (MRSA) and Pseudomonas aeruginosa. Furthermore, slow release of copper ions from the hydrogel facilitated wound healing by promoting cell migration, collagen deposition and angiogenesis. Additionally, the PCC hydrogel possessed excellent biocompatibility and hemostatic properties. The practical effectiveness of PCC hydrogel in addressing bacterial infections and facilitating wound healing was verified using a mouse model of MRSA-induced wound infections. Overall, this work presents a simple yet efficient multifunctional hydrogel platform that integrates antibacterial activity, promotion of wound healing, and hemostasis for managing bacteria-associated wounds.

Mesenchymal stem cells (MSCs) are naturally-derived regenerative materials that exhibit significant potential in regenerative medicine. Previous studies have demonstrated that MSCs-based therapy can improve heart function in ischemia-injured hearts, offering an exciting therapeutic intervention for myocardial ischemic infarction, a leading cause of worldwide mortality and disability. However, the efficacy of MSCs-based therapies is significantly disturbed by the myocardial microenvironment, which undergoes substantial changes following ischemic injury. After the ischemic injury, blood vessels become obstructed and damaged, and cardiomyocytes experience ischemic conditions. This activates the hypoxia-induced factor 1 (HIF-1) pathway, leading to the rapid production of several cytokines and chemokines, including vascular endothelial growth factor (VEGF) and stromal-derived factor 1 (SDF-1), which are crucial for angiogenesis, cell migration, and tissue repair, but it is not sustainable. MSCs respond to these cytokines and chemokines by homing to the injured site and participating in myocardial regeneration. However, the deteriorated microenvironment in the injured myocardium poses challenges for cell survival, interacting with MSCs, and constraining their homing, retention, and migration capabilities, thereby limiting their regenerative potential. This review discusses how the deteriorated microenvironment negatively affects the ability of MSCs to promote myocardial regeneration. Recent studies have shown that optimizing the microenvironment through the promotion of angiogenesis can significantly enhance the efficacy of MSCs in treating myocardial infarction. This approach harnesses the full therapeutic potential of MSCs-based therapies for ischemic heart disease.

CRISPR/Cas9 genome editing is a rapidly advancing technology that has the potential to accelerate research and development in a variety of fields. However, manual genome editing processes suffer from limitations in scalability, efficiency, and standardization. The implementation of automated systems for genome editing addresses these challenges, allowing researchers to cover the increasing need and perform large-scale studies for disease modeling, drug development, and personalized medicine. In this study, we developed an automated CRISPR/Cas9-based genome editing process on the StemCellFactory platform. We implemented a 4D-Nucleofector with a 96-well shuttle device into the StemCellFactory, optimized several parameters for single cell culturing and established an automated workflow for CRISPR/Cas9-based genome editing. When validated with a variety of genetic backgrounds and target genes, the automated workflow showed genome editing efficiencies similar to manual methods, with indel rates of up to 98%. Monoclonal colony growth was achieved and monitored using the StemCellFactory-integrated CellCelector, which allowed the exclusion of colonies derived from multiple cells or growing too close to neighbouring colonies. In summary, we demonstrate the successful establishment of an automated CRISPR/Cas9-based genome editing process on the StemCellFactory platform. The development of such a standardized and scalable automated CRISPR/Cas9 system represents an exciting new tool in genome editing, enhancing our ability to address a wide range of scientific questions in disease modeling, drug development and personalized medicine.

Introduction: Early diagnosis of oral squamous cell carcinoma can greatly improve treatment success rate and patient survival. Although Optical Coherence Tomography (OCT) based Angiography (OCTA) is a promising in vivo technique in oral imaging, there is a need for objective assessment of oral microvasculature.

Methods: This study aimed to demonstrate a comprehensive methodology of quantitative assessing OCTA intraoral scanning results to provide measurable, reproducible data and to avoid subjective visual interpretations. Data were collected from 37 healthy subjects in total across four intraoral sites-buccal mucosa (n = 32), labial mucosa (n = 24), floor of the mouth (n = 13), and hard palate (n = 8)-using a non-invasive swept-source OCT system. Four quantitative metrics-vessel area density, vessel skeleton density, vessel diameter index, and a newly proposed weighted Tortuosity Index-were used to assess OCTA images in oral applications.

Results: The quadruple quantitative assessment's repeatability was evaluated to be reliable. Analysis of a benign ulcer case revealed differences in these metrics compared to healthy cases.

Discussion/conclusion: In conclusion, we demonstrated a comprehensive method to quantify microvasculature in the oral cavity, showing considerable promise for early diagnosis and clinical management of oral diseases.

D-Phenyllactic acid (D-PLA) is a potent antimicrobial typically synthesized through chemical methods. However, due to the complexity and large pollution of these reactions, a simpler and more eco-friendly approach was needed. In this study, a strain for D-PLA biosynthesis was constructed, but the efficiency was restricted by the activity of D-lactate dehydrogenase (DLDH). To address this issue, a DLDH mutant library was constructed and the Surface-Enhanced Raman Spectroscopy (SERS) was employed for the precise quantification of D-PLA at the single-cell level. The TB24 mutant exhibited a significant improvement in D-PLA productivity and a 23.03-fold increase in enzymatic activity, which was attributed to the enhanced hydrogen bonding and increased hydrophobicity within the substrate-binding pocket. By implementing multi-level optimization strategies, including the co-expression of glycerol dehydrogenase (GlyDH) with DLDH, chassis cell replacement, and RBS engineering, a significant increase in D-PLA yields was achieved, reaching 128.4 g/L. This study underscores the effectiveness of SERS-based microdroplet high-throughput screening (HTS) in identifying superior mutant enzymes and offers a strategy for large-scale D-PLA biotransformation.

The perpetual release of natural/synthetic pollutants into the environment poses major risks to ecological balance and human health. Amongst these, contaminants of emerging concern (CECs) are characterized by their recent introduction/detection in various niches, thereby causing significant hazards and necessitating their removal. Pharmaceuticals, plasticizers, cyanotoxins and emerging pesticides are major groups of CECs that are highly toxic and found to occur in various compartments of the biosphere. The sources of these compounds can be multipartite including industrial discharge, improper disposal, excretion of unmetabolized residues, eutrophication etc., while their fate and persistence are determined by factors such as physico-chemical properties, environmental conditions, biodegradability and hydrological factors. The resultant exposure of these compounds to microbiota has imposed a selection pressure and resulted in evolution of metabolic pathways for their biotransformation and/or utilization as sole source of carbon and energy. Such microbial degradation phenotype can be exploited to clean-up CECs from the environment, offering a cost-effective and eco-friendly alternative to abiotic methods of removal, thereby mitigating their toxicity. However, efficient bioprocess development for bioremediation strategies requires extensive understanding of individual components such as pathway gene clusters, proteins/enzymes, metabolites and associated regulatory mechanisms. "Omics" and "Meta-omics" techniques aid in providing crucial insights into the complex interactions and functions of these components as well as microbial community, enabling more effective and targeted bioremediation. Aside from natural isolates, metabolic engineering approaches employ the application of genetic engineering to enhance metabolic diversity and degradation rates. The integration of omics data will further aid in developing systemic-level bioremediation and metabolic engineering strategies, thereby optimising the clean-up process. This review describes bacterial catabolic pathways, genetics, and application of omics and metabolic engineering for bioremediation of four major groups of CECs: pharmaceuticals, plasticizers, cyanotoxins, and emerging pesticides.

Articular cartilage defects often involve damage to both the cartilage and subchondral bone, requiring a scaffold that can meet the unique needs of each tissue type and establish an effective barrier between the bone and cartilage. In this study, we used 3D printing technology to fabricate a tri-phasic scaffold composed of PLA/PCL-PLGA/Mg(OH)₂, which includes a cartilage layer, an osteochondral interface, and a bone layer. The scaffold was filled with Velvet antler polypeptides (VAP), and its characterization was assessed using compression testing, XRD, FTIR, SEM, fluorescence microscopy, and EDS. In vitro investigation demonstrated that the scaffold not only supported osteogenesis but also promoted chondrogenic differentiation of fibrocartilage stem cells (FCSCs). n vivo experiments showed that the tri-phasic PLA/PCL-PLGA/Mg(OH)₂-VAP scaffold together with FCSC, when transplanted to animal models, increased the recovery of osteochondral defects. Those results demonstrate the promising future of illustrated tri-phasic PLA/PCL-PLGA/Mg(OH)₂-VAP scaffold loaded with FCSCs as a new bone and cartilage tissue engineering approach for osteochondral defects treatment.