Frontiers in Bioengineering and Biotechnology最新文献

Our previous article entitled "Proteomics and its applications in breast cancer", proposed a Breast Cancer Continuum Concept (BCCC), including a Breast Cancer Cell Continuum Concept as well as a Breast Cancer Proteomic Continuum Concept. Breast cancer-on-chip (BCoC), breast cancer liquid biopsy-on-chip (BCLBoC), and breast cancer metastasis-on-chip (BCMoC) models successfully recapitulate and reproduce in vitro the principal mechanisms and events involved in BCCC. Thus, BCoC, BCLBoC, and BCMoC platforms allow for multiple cell lines co-cultivation to reproduce BC hallmark features, recapitulating cell proliferation, cell-to-cell communication, BC cell-stromal crosstalk and stromal activation, effects of local microenvironmental conditions on BC progression, invasion/epithelial-mesenchymal transition (EMT)/migration, intravasation, dissemination through blood and lymphatic circulation, extravasation, distant tissues colonization, and immune escape of cancer cells. Moreover, tumor-on-chip platforms are used for studying the efficacy and toxicity of chemotherapeutic drugs/nano-drugs or nutraceuticals. Therefore, the aim of this review is to summarize and analyse the main bio-medical roles of on-chip platforms that can be used as powerful tools to study the metastatic cascade in BC. As future direction, integration of tumor-on-chip platforms and proteomics-based specific approaches can offer important cues about molecular profile of the metastatic cascade, alowing for novel biomarker discovery. Novel microfluidics-based platforms integrating specific proteomic landscape of human milk, urine, and saliva could be useful for early and non-invasive BC detection. Also, risk-on-chip models may improve BC risk assessment and prevention based on the identification of biomarkers of risk. Moreover, multi-organ-on-chip systems integrating patient-derived BC cells and patient-derived scaffolds have a great potential to study BC at integrative level, due to the systemic nature of BC, for personalized and precision medicine. We also emphasized the strengths and weaknesses of BCoC and BCMoC platforms.

Introduction: Decellularized uterine extracellular matrix has emerged as a pivotal focus in the realm of biomaterials, offering a promising source in uterine tissue regeneration, research on disease diagnosis and treatments, and ultimately uterine transplantation. In this study, we examined various protocols for decellularizing porcine uterine tissues, aimed to unravel the intricate dynamics of DNA removal, bioactive molecules preservation, and microstructural alterations.

Methods: Porcine uterine tissues were treated with 6 different, yet rigorously selected and designed, protocols with sodium dodecyl sulfate (SDS), Triton^® X-100, peracetic acid + ethanol, and DNase I. After decellularization, we examined DNA quantification, histological staining (H&E and DAPI), glycosaminoglycans (GAG) assay, scanning electron microscopy (SEM), Fourier-transform infrared spectroscopy (FT-IR), X-ray photoelectron spectroscopy (XPS), and Thermogravimetric Analysis (TGA).

Results: A comparative analysis among all 6 protocols was conducted with the results demonstrating that all protocols achieved decellularization; while 0.1% SDS + 1% Triton^® X-100, coupled with agitation, demonstrated the highest efficiency in DNA removal. Also, it was found that DNase I played a key role in enhancing the efficiency of the decellularization process by underscoring its significance in digesting cellular contents and eliminating cell debris by 99.79% (19.63 ± 3.92 ng/mg dry weight).

Conclusions: Our findings enhance the nuanced understanding of DNA removal, GAG preservation, microstructural alteration, and protein decomposition in decellularized uterine extracellular matrix, while highlighting the importance of decellularization protocols designed for intended applications. This study along with our findings represents meaningful progress for advancing the field of uterine transplantation and related tissue engineering/regenerative medicine.

Introduction: Woven bone, a heterogeneous and temporary tissue in bone regeneration, is remodeled by osteoblastic and osteoclastic activity and shaped by mechanical stress to restore healthy tissue properties. Characterizing this tissue at different length scales is crucial for developing micromechanical models that optimize mechanical parameters, thereby controlling regeneration and preventing non-unions.

Methods: This study examines the temporal evolution of the mechanical properties of bone distraction callus using nanoindentation, ash analysis, micro-CT for trabecular microarchitecture, and Raman spectroscopy for mineral quality. It also establishes single- and two-parameter power laws based on experimental data to predict tissue-level and bulk mechanical properties.

Results: At the macro-scale, the tissue exhibited a considerable increase in bone fraction, controlled by the widening of trabeculae. The Raman mineral-to-matrix ratios increased to cortical levels during regeneration, but the local elastic modulus remained lower. During healing, the tissue underwent changes in ash fraction and in the percentages of Calcium and Phosphorus. Six statistically significant power laws were identified based on the ash fraction, bone fraction, and chemical and Raman parameters.

Discussion: The microarchitecture of woven bone plays a more significant role than its chemical composition in determining the apparent elastic modulus of the tissue. Raman parameters were demonstrated to provide more significant power laws correlations with the micro-scale elastic modulus than mineral content from ash analysis.

Cell and gene therapy (CGT) is a field of therapeutic medicine that aims to treat, prevent, and cure diseases using engineered cells (stem cells, immune cells, and differentiated adult or fetal cells), vectors [Adeno Associated Virus (AAV), Adeno Virus (AV), Herpes Simplex Virus (HSV), Baculo Virus (BV), Lenti Virus (LV), Retro Virus (RV), etc.], and other carriers [non-viral vectors, virus-like particles (VLP), Lipid Nano-Particles (LNP), etc.]. Among viral CGT vectors, adeno-associated viruses and lentiviruses (AAV and LV) are the most widely applied vector platforms. The presence of non-functional (empty or non-infectious) vectors that carry null or partial genes in the final drug product is classified as an impurity by the FDA. These impurities impair dosage accuracy and induce non-specific immunogenicity and variability in drug efficacy. These non-functional viral vectors in the drug product need to be elucidated following International Conference on Harmonization (ICH) guidelines for clinical manufacturing of the final drug product. This article showcases an ion-exchange chromatography (IEX) high-resolution method supporting ICH guidelines using commercially available AAV8 filled and empty capsids as reference standards. Our method successfully separated empty to full capsids with a resolution of 15 and sustained a linearity greater than 0.98 even under a wide range of empty or full viral particle concentrations (E+9 to E+13 vp/mL), which is an upgrade to other IEX capsid separation methods. The medium-throughput capacity and shorter sample processing time improve testing efficiency and save costs while delivering quality as value. The discussed method is a reliable and reproducible platform to precisely evaluate the presence of non-functional viral particles in AAV8 samples. Aligned with other orthogonal results, the method is a powerful tool to improve the quality of rAAV analytics.

Object: In recent years, dielectrophoresis has become widely recognized as a highly suitable method for creating good tools for particle separation, with significant successes achieved in a variety of areas.

Method: Expanding upon this, we adopted a semiconductor CMOS process, instead of a MEMS process, which allowed for the following: 1) wire insulation to mitigate Joule heat and prevent thermal fluctuation interference with the dielectrophoretic force; 2) isolation of harmful materials from biological samples, making the chip biocompatible; and, 3) the ability to employ nano-electrodes capable of generating a stronger electric field than conventional electrodes, thus allowing chip capture at lower voltages. Additionally, our chip is scalable, enabling multiplied throughput based on sample processing requirements.

Results and dissusion: These features make our chip more widely applicable and suitable for capturing bacteria and sperm. In this study, we focused on optimizing the parameters of dielectrophoresis and employed 3-D protruding TiN nano-electrode arrays to facilitate the capture of Escherichia coli and boar sperms. The experimental data demonstrates that the capture efficiency of this chip for E. coli was approximately 79.25% ± 2.66%, and the highest capture efficiency for sperms was approximately 39.2% ± 3.9%.

This comprehensive review examines the latest developments in improving the blood compatibility of hemoperfusion adsorbents. By leveraging advanced coating and modification techniques, including albumin-collodion, cellulose, hydrogel, and heparin coatings, notable enhancements in blood compatibility have been achieved across diverse adsorbent types, such as carbon-based, resin-based, and polysaccharide-based materials. Despite promising laboratory results, the intricate manufacturing processes and elevated costs present significant challenges for broad clinical application. Therefore, future endeavors should focus on cost-benefit analysis, large-scale production strategies, in-depth exploration of blood-material interactions, and innovative technologies to propel the development of safer and more effective blood purification therapies.

Synthetic biology is an interdisciplinary field that brings together engineering and biology concepts alongside the arts and social sciences to develop solutions to pressing problems in our world. The education of students entering this field has relied on a diverse set of pedagogical methods to accomplish this goal. One non-profit group, iGEM-the International Genetically Engineered Machine competition, has been a driver of students' awareness of synthetic biology for the last 20 years giving many young researchers their first experience in the field of synthetic biology. Dissemination of synthetic biology concepts by iGEM has occurred through several programs including a webinar series started during the 2020 COVID pandemic. The iGEM webinar series successfully engaged students by taking inspiration from synthetic biology programs in Europe, North America, and Asia that had themselves evolved alongside iGEM. The webinar designers modeled the content after their experiences in iGEM as well as their academic courses, pedagogy, and mentoring experiences. This series has produced globally accessible pedagogy for both technical synthetic biology knowledge and the communication skills necessary to build and communicate synthetic biology projects. The hope is that this series functions as a lasting blueprint that can be used by future educators in synthetic biology and other disciplines to reduce barriers that students face when attempting to enter cutting edge fields.

Extracellular vesicles (EVs) play an important role in normal life activities and disease treatment. In recent years, there have been abundant relevant studies focusing on EVs for cancer therapy and showing good performance on tumor inhibition. To enhance the effectiveness of EVs, EV analogs have been developed. This review summarizes the classification, origin, production, purification, modification, drug loading and cancer treatment applications of EVs and their analogs. Also, the characteristics of technologies involved are analyzed, which provides the basis for the development and application of biogenic vesicle-based drug delivery platform for cancer therapy. Meanwhile, challenges in translating these vesicles into clinic, such as limited sources, lack of production standards, and insufficient targeting and effectiveness are discussed. With ongoing exploration and clinical studies, EV-based drugs will make great contributions to cancer therapy.

Background: Mandibular defects pose significant challenges in reconstructive surgery, and scaffold materials are increasingly recognized for their potential to address these challenges. Among various scaffold materials, Beta-tricalcium phosphate (β-TCP) is noted for its exceptional osteogenic properties. However, improvements in its biodegradation rate and mechanical strength are essential for optimal performance.

Methods: In this study, we developed a novel β-TCP-based scaffold, CFBB, by calcining fetal bovine cancellous bone. To enhance its properties, we modified CFBB with Chitosan (CS) and Zinc (Zn), creating three additional scaffold materials: CFBB/CS, CFBB/Zn²⁺, and CFBB/Zn²⁺/CS. We conducted comprehensive assessments of their physicochemical and morphological properties, degradation rates, biocompatibility, osteogenic ability, new bone formation, and neovascularization both in vitro and in vivo.

Results: Our findings revealed that all four materials were biocompatible and safe for use. The modifications with CS and Zn²⁺ significantly improved the mechanical strength, osteogenic, and angiogenic properties of CFBB, while concurrently decelerating its resorption rate. Among the tested materials, CFBB/Zn²⁺/CS demonstrated superior performance in promoting bone regeneration and vascularization, making it a particularly promising candidate for mandibular reconstruction.

Conclusion: The CFBB/Zn²⁺/CS scaffold material, with its enhanced mechanical, osteogenic, and angiogenic properties, and a controlled resorption rate, emerges as a highly effective alternative for the repair of oral mandible defects. This study underscores the potential of combining multiple bioactive agents in scaffold materials to improve their functionality for specific clinical applications in bone tissue engineering.