Biofabrication最新文献

2D Ti₃C₂T_x(MXene) is attracting significant attention in tissue engineering. The incorporation of these promising materials into conventional scaffolds remains challenging, particularly with physicochemical properties compatible with biological systems. Melt electrowriting (MEW) has emerged as a powerful additive manufacturing technique for biofabrication of customized three-dimensional (3D) scaffolds composed of bioactive materials. This study introduces MEW of 2D MXene and polycaprolactone (PCL) nanocomposite scaffolds for tissue engineering applications. First, Ti₃C₂T_xwas functionalized using (3-aminopropyl) triethoxysilane (referred to asf-MXene) to obtain a blended nanocomposite in PCL matrix (referred to as MX/PCL). Fourier transform infrared spectroscopy revealed the nanocomposite composition. X-ray diffraction analysis showed the reduced crystallinity in PCL after incorporation off-MXene. Differential scanning calorimetry helped to establish the optimal MEW parameters. Thermogravimetric analysis conducted on nanocomposites containing 0.1, 0.5, and 1% (w/w)f-MXene showed the thermal stability of MXene during the MEW process. The extrudability and printability of the nanocomposites with varying concentrations was demonstrated using MEW in 0-90-degree mesh scaffolds with fine filament dimensions. Scanning electron microscopy and Energy-dispersive x-ray spectroscopy mapping showed the shape fidelity, printing accuracy, and structural integrity of 3D MEW scaffolds with uniform distribution off-MXene, respectively. Further characterization showed the concentration-dependent enhancement in hydrophilicity and compressive modulus and yield strength of scaffolds upon integration off-MXene. Atomic force microscopy analysis demonstrated that the topography of the 3D MEW MX/PCL scaffolds changed compared to the pristine PCL and the roughness of the surfaces increased as the concentration of thef-MXene increased. Accelerated degradation tests demonstrated that increasing filler concentration in the reinforced scaffolds progressively delayed degradation compared to the control. Thein vitrocharacterization showed the adherence of MC3T3-E1 preosteoblast cells on MX/PCL scaffolds and their enhanced osteogenic differentiation. The findings indicate that 3D printed MX/PCL nanocomposite scaffolds have significant potential as mechanically robust scaffolds with controlled degradation rate and cytocompatibility for tissue regeneration, with properties tunable for specific applications.

The intricate three dimensional architecture at different spatial length scales affects the functionality and growth performance of immobilized photosynthesizing cells in biofilms and bioprinted constructs. Despite the tremendous potential of 3D bioprinting in precisely defining sample heterogeneity and composition in spatial context, cell metabolism is mostly measured in media surrounding the constructs or by destructive sample analyzes. The exploration and application of non-invasive techniques for monitoring physico-chemical microenvironments, growth and metabolic activity of cells in 3D printed constructs is thus in strong demand. Here, we present a pipeline for the fabrication of 3D bioprinted microalgal constructs with a functionalized gelatin methacryloyl-based bioink for imaging O₂dynamics within bioprinted constructs, as well as their characterization using various, non-invasive functional imaging techniques in concert with numerical simulation of their photophysiological performance. This fabrication, imaging and simulation pipeline now enables investigation of the effect of structure and composition on photosynthetic efficiency of bioprinted constructs with microalgae or cyanobacteria. It can facilitate designing efficient construct geometries for enhanced light penetration and improved mass transfer of nutrients, CO₂or O₂between the 3D printed construct and the surrounding medium, thereby providing a mechanistic basis for the design of more efficient artificial photosynthetic systems.

The advent of 3D bioprinting has revolutionized tissue engineering and regenerative medicine. Today, tissues of single cell type can be fabricated with extreme resolution and printing fidelity. However, the ultimate functionality of the desired tissue is limited, due to the absence of a multicellular population and diversity in micro-environment distribution. Currently, 3D bioprinting technologies are facing challenges in delivering multiple cells and biomaterials in a controlled fashion. The use of interchangeable syringe-based systems has often favoured the delamination between interfaces, greatly limiting the fabrication of interconnected tissue constructs. Microfluidic-assisted 3D bioprinting platforms have been found capable of rescuing the fabrication of tissue interfaces, but often fails to guarantee printing fidelity, cell density control and compartmentalization. Herein, we present the convergence of microfluidic and 3D bioprinting platforms into a deposition system capable of harnessing a microfluidic printhead for the continuous rapid fabrication of interconnected functional tissues. The use of flow-focusing and passive mixer printhead modules allowed for the rapid and dynamic modulation of fibre diameter and material composition, respectively. Cells were compartmentalized into discrete three-dimensional layers with defined density patterns, confirming the punctual control of the presented microfluidic platform in arranging cells and materials in 3D.In ovoandin vivostudies demonstrated the seminal functionality of 3D bioprinted constructs with patterned vascular endothelial growth factor and transforming growth factor-β1 (TGF-β1), respectively. This, in turn, facilitated the simulation of diverse cellular environments and proliferation pathways within a single construct, which is currently unachievable with conventional 3D bioprinting techniques, offering new opportunities for the fabrication of functionally graded systems and physiologically-relevant skeletal tissue substitutes.

Tumor organoids that can accurately recapitulate the pathophysiological characteristics of original tumor are urgently needed for personalized therapy. However, there are few published studies on patient-derived renal cell carcinoma (RCC) heterogeneous organoids for drug testing to account for patient-specific heterogeneous clinical responses, which has significantly impeded research in the field. Traditional RCC organoid technologies involving matrigel droplets require intensive manual manipulation and are hampered by variability, functional immaturity, low throughput, and limited scale. Here, we applied extrusion-based high-throughput bioprinter to rapidly generate heterogeneous RCC organoids with uniform size, realizing batch automated stable construction and quality control. Bioprinted RCC organoids reserved the pathological morphology and gene mutation/expression characteristics of original tumor and demonstrate interorganoid and interpatient heterogeneity even after long-term cultivation, which are suitable for preclinical patient-specific drug screening testing. Finally, we created multicellular assembloids by reconstituting RCC aggregates with stromal components to generate an organized architecture within vivo-like vascular morphology and spatial tumor microenvironment heterogeneity. Thus, we have demonstrated the wide-ranging biomedical utility of bioprinted organoids in furthering our understanding of the physiological mechanisms of tumors and the development of personalized treatment methods.

Myalgic encephalomyelitis/chronic fatigue syndrome (ME/CFS) and Long Covid-19 (LC-19) are complex conditions with no diagnostic markers or consensus on disease progression. Despite extensive research, noin vitromodel exists to study skeletal muscle wasting, peripheral weakness, or potential therapies. We developed 3Din vitroskeletal muscle tissues to map muscle adaptations to patient sera over time. Short exposures (48 H) to patient sera led to a significant reduction in muscle contractile strength. Transcriptomic analysis revealed the upregulation of protein translation, glycolytic enzymes, disturbances in calcium homeostasis, hypertrophy, and mitochondrial hyperfusion. Structural analyses confirmed myotube hypertrophy and elevated mitochondrial oxygen consumption In ME/CFS. While muscles initially adapted by increasing glycolysis, prolonged exposure (96-144 H) caused muscle fragility and weakness, with mitochondria fragmenting into a toroidal conformation. We propose that skeletal muscle tissue in ME/CFS and LC-19 progresses through a hypermetabolic state, leading to severe muscular and mitochondrial deterioration. This is the first study to suggest such transient metabolic adaptation.

In this study, a multi breast cell-on-a-chip system was developed to recapitulate human-like breast cancer microenvironments by simultaneously co-culturing normal (MCF-10 A) and breast cancer cell lines (MCF-7 and MDA-MB-231) and to confirm the potential of plasma-activated PBS (P-PBS) containing various reactive oxygen species (ROS), as selective breast cancer remedial agent. The developed chip not only provided a 3D microenvironment supporting cell survival and interactions due to the use of gelatin as an extracellular matrix but also facilitated homogeneous growth and interactions between the different breast cell lines. Moreover, selective breast cancer apoptosis was investigated by real-time image analysis after treating cells with P-PBS on the developed chip; IC₅₀values for MCF-7 and MDA-MB-231 were 390.50 and 600.75μM, respectively. In addition, the breast cancer cell apoptosis pathway was found to be related to the ROS-activated intrinsic apoptosis pathway. The developed multi breast cells-on-a-chip platform could be used as a bio-platform that complements the limitations of existingin vitromodels. The chip accurately reproduces cell-to-cell interactions by providing a tumor microenvironment based on the co-culture of heterogeneous cancer and normal cells and is expected to contribute meaningfully to breast cancer research and customized treatment development.

Over the past decade, three-dimensional (3D) bioprinting has made significant progress, transforming into a key innovation in tissue engineering. Despite the early strides, critical challenges remain in 3D bioprinting that must be addressed to accelerate clinical translation. In particular, there is still a long way to go before functionally-mature, clinically-relevant tissue equivalents are developed. Current limitations range from the sub-optimal bioink properties and degree of biomimicry of bioprintable architectures, to the lack of stem/progenitor cells for massive cell expansion, and fundamental knowledge regardingin vitroculturing conditions. In addition to these problems, the absence of guidelines and well-regulated international standards is creating uncertainty among the biofabrication community stakeholders regarding the reliable and scalable production processes. This review aims at exploring the latest developments in 3D bioprinting approaches, including various additive manufacturing techniques and their applications. A thorough discussion of common bioprinting techniques and recent progresses are compiled along with notable recent studies. Later we discuss the current challenges in clinical application of 3D bioprinting and the major bottlenecks in the commercialization of 3D bioprinted tissue equivalents, including the longevity of bioprinted organs, meeting biomechanical requirements, and the often underrated ethical and legal aspects. Amidst the progress of regulatory efforts for regenerative medicine, we also present an overview of the current regulatory concerns which should be taken into account to translate bioprinted tissues into clinical practice. At last, this review emphasizes future directions in 3D bioprinting that includes the transformative ideas such as bioprinting in microgravity and the integration of artificial intelligence. The study concludes with a discussion on the need for collaborative efforts in resolving the technical and regulatory constraints to improve the quality, reliability, and reproducibility of bioprinted tissue equivalents to ultimately accomplish their successful clinical implementation.

The dysregulated and fibrotic tumor microenvironment of hepatocellular carcinoma delays diagnosis and presents many complex signals that drive disease progression. To better recapitulate this microenvironment, we have enhanced our established protein microarray platform by integrating design of experiments (DoEs) methodology with high-throughput cell microarray screening. This innovative approach systematically interrogates the intricate roles of matrix stiffness (spanning healthy and fibrotic conditions), extracellular matrix (ECM) composition, and protein concentration, while simultaneously examining their interdependent interactions. By leveraging DoE principles, we were able to explore 117 unique microenvironments on a single microscope slide, ultimately generating a comprehensive dataset of 234 different microenvironments without compromising statistical rigor. Our enhanced screening system enabled the identification of unique microenvironmental interactions critically significant in dictating cellular responses, including adhesion, survival, proliferation, epithelial-to-mesenchymal transition, and drug resistance markers. Utilizing advanced statistical techniques such as linear models and principal component analysis, we characterized phenotypic clusters defined by precise microenvironmental cues. This work presents a robust, high-throughput microarray screening system that comprehensively explores the contributions of nine physiologically relevant ECM proteins and matrix stiffness in modulating cellular behavior and disease progression through a methodologically sophisticated and statistically sound approach.

This study addresses the challenges of tracking cell-mediated biodegradation in cartilage tissue engineering, where hydrogels and scaffolds play a crucial role in providing structural support and promoting tissue regeneration. This research area has been rarely studied, offering potential insights into bridging the gap betweenin vitroandin vivoconditions for real-time monitoring of tissue regeneration alongside biodegradation. We developed dual-labeled hydrogel/scaffold composites for real-time monitoring of scaffold degradation in response to cell activity. Gelatin methacryloyl (GelMA) hydrogels are extensively explored for cartilage tissue engineering, albeit concerns remain regarding their mechanical properties under load-bearing conditions. To address this, a hydrogel/scaffold composite system was employed in this study, where a poly (ϵ-caprolactone) (PCL) hex prism edge structure acts as a scaffold to support the cell-laden GelMA hydrogel. Fluorophore labeling of GelMA and PCL facilitated non-invasive monitoring of the hydrogel/scaffold composite biodegradation under cell proliferation conditions. Initially, the behavior of fluorescent-tagged Hydrogel/Scaffold was examined under accelerated degradation conditions. Subsequently, human adipose-derived mesenchymal stem cells loaded into fluorescent-labeled hydrogel/scaffolds were evaluated for their biocompatibility potential and chondrogenesis. Results demonstrated a correlation between the loss of fluorescence from the hydrogel/scaffold degradation, accompanied by extracellular matrix accumulation. The fluorescently labeled hydrogel/scaffold holds promising application for cartilage tissue engineering, offering the capability to monitor biodegradation using high-throughput and contactless techniques.

Wearable devices have emerged as powerful tools for continuous, real-time health monitoring, enabling the detection of biochemical markers in sweat, tears, saliva, and interstitial fluid. However, existing wearable materials are hindered by limited chemical functionality, static sensing capabilities, and insufficient adaptability to dynamic physiological conditions, which restrict their current impact in precision medicine. Recent advancements have focused on integrating genetic engineering and synthetic biology into wearable platforms, resulting in genetically programmable biointerfaces that enhance specificity, responsiveness, and functional versatility in clinical and personalized healthcare settings. Current applications of these bioengineered devices include real-time monitoring of pathogens, hormones, therapeutic drug levels, and physiological behaviors, offering superior precision and adaptability compared to traditional wearable technologies. This review highlights two key engineering approaches driving this field: genetically modified living cells and cell-free synthetic biology systems. While promising, several challenges still limit broader clinical adoption, including biosafety concerns, the instability of biological components, and translational hurdles. Addressing these challenges requires progress in biocompatibility, controlled gene expression, and durable wearable materials. Looking ahead, future research should aim to integrate these biointerfaces with implantable and smart therapeutic systems, develop autonomous biosensors with self-regulatory functions, and further expand their use in personalized medicine and real-time disease management. By bridging genetic programming with wearable diagnostics, these innovations are laying the groundwork for next-generation biohybrid systems designed to advance precision healthcare.