Biofabrication最新文献

The drug development process in cancer faces significant challenges due to high failure rates in translational studies despite promising in vitro results. Additionally, conventional animal models exhibit inherent limitations and ethical concerns, constraining their relevance to cancer studies. Recognizing the pivotal role of the tumor microenvironment (TME) on cancer development and treatment outcome, recent advancements in 3D microfluidic devices and tumor-on-a-chip models enabled researchers to explore the TME with enhanced accuracy and reliability, yielding novel insights. Notably, the emergence of physiological tumor models, particularly 3D models such as organoids derived from human tissues, provides a more accurate representation of in vivo tumor features. Moreover, 3D tumor models hold promise for diverse applications, including highthroughput drug testing, disease modeling, and regenerative medicine. Meanwhile, combining artificial intelligence (AI) with patient-derived tumor organoids has become a key strategy in predictive oncology and personalized cancer treatment. Furthermore, incorporating quantitative systems pharmacology (QSP) and physiologically based pharmacokinetic (PBPK) modeling, and pharmacokinetics/pharmacodynamics (PK/PD) analysis with generative artificial intelligence (Gen-AI) has revolutionized predictive oncology by enabling precise simulations of drug interactions and patient-specific responses, thereby enhancing the predictive accuracy of personalized cancer treatments. These advanced methodologies harness the power of AI algorithms to analyze intricate datasets derived from patient-specific tumor organoids. Moreover, the predictive modeling capabilities of generative AI facilitate the development of personalized treatment strategies customized for each patient, thereby revolutionizing oncology practice. This review explores the synergistic impact of tumor-on-a-chip models, organoids derived from patient tumors, and generative AI. Together, these technologies mark a significant advancement in precision medicine, offering promising chances to improve therapeutic effectiveness and treatment outcomes in cancer care.

Thrombosis is a leading cause of cardiovascular morbidity and mortality, driven by platelet-mediated mechanisms common across distinct vascular environments. However, the dynamical behavior of platelets during thrombogenesis remains poorly understood due to the lack of a comprehensive analytical framework. Here we present a physiologically relevant, imaging-integrated thrombosis-on-a-chip platform that enables real-time quantitative analysis of platelet dynamics during thrombogenesis under arterial, venous, and cancer-associated conditions. The system incorporates endothelialized 3D-printed vascular geometries into a closed-loop whole-blood perfusion circuit that replicates native hemodynamic and cellular microenvironments. Multimodal imaging captures the spatiotemporal evolution of thrombus formation and shows how hydrodynamic forces, endothelial dysfunction, and tumor-derived factors drive distinct thrombotic signatures. Notably, platelet-endothelium adhesion and circulating platelet aggregation are identified as mechanistically distinct yet closely linked processes, each uniquely modulated by vascular context. This platform offers a robust framework for dissecting thrombogenesis and advancing antithrombotic and cancer-associated thrombosis research.

Large bone defects are usually treated with autografts, allografts, synthetic materials, or bioactive ceramics. Each of these options has limitations, including donor site issues, limited availability, and poor fit for individual patient needs. Bioactive glasses have the potential to promote bone growth but often lack sufficient strength for clinical use. This study investigated the possibility of using 3D-printed bone grafts made from polylactic acid (PLA), polycaprolactone (PCL), and borate bioactive glass (BBG, 13-93B3) to create customized and strong grafts. We created composite filaments (PLA/BBg and PCL/BBg; 50 wt% polymer and 50 wt% BBg) as well as pure PLA and PCL filaments. We then tested for the compatibility of these materials with cells and living tissue, their mechanical properties, and the ease of printing. Tests with mesenchymal stem cells revealed increased metabolic activity of approximately 15.4% for PCL/BBg and 20.7% for PLA/BBg and higher alkaline phosphatase activity of approximately 20.8% and 35.4%, respectively, on day 7, which indicates better early bone formation. The constructs made from PCL/BBg had mineral contents similar to those of natural bone and were easy to print via fused deposition modeling. They produced porous structures with a compressive strength of approximately 9 MPa and a modulus of 168 MPa that matched human mandibular trabecular bone.In vivotests on a rabbit model with premaxillary defects over 14 weeks revealed better bone healing in the PCL/BBg-treated areas, with no signs of inflammation or toxicity. These results indicate that PCL/BBg composite filaments could be effective for making patient-specific, biodegradable bone grafts that have precise structures and strong mechanical properties. More studies involving larger animals are needed to push this research toward clinical use.

Large bone defects present a major clinical challenge, exceeding the body's natural regenerative capacity. In this study, we investigated the physicochemical and biological properties of a novel cross-linked collagen-hydroxyapatite (Coll-HA-XL) biomaterial ink designed for additive manufacturing of scaffolds for bone tissue engineering. The biomaterial ink was developed through three stages: initially as a hydrogel, then molded and freeze-dried into disk-shaped forms, and finally tailored into three-dimensional (3D)-printed scaffolds subjected to subsequent freeze-drying. To optimize the ink, we systematically varied the HA proportions and the sequence of HA incorporation and cross-linking. The composite materials were then 3D-printed into scaffolds by a direct ink writing method, and were seeded with primary human osteoblasts (hOBs). The introduction of HA and subsequent collagen cross-linking induced a significantly increased storage modulus and thermal stability of the material, when compared with the non-crosslinked, HA-containing controls. Biocompatibility of the materials was assessed by hOB cultures, and Coll-HA-XL induced higher alkaline phosphatase (ALP) and lactate dehydrogenase (LDH) activity when compared to the non-crosslinked control. After four weeks of culture on 3D-printed Coll-HA-XL scaffolds, high ALP and LDH activities and osteocalcin staining of hOB indicated robust osteoblastic differentiation. Our findings show that a crosslinked, collagen-based biomaterial ink supplemented with HA is suitable for direct ink writing of scaffolds tailored for bone tissue engineering.

A central hurdle in spinal cord injury (SCI) therapy involves not only reconstructing neural pathways but also overcoming the detrimental inflammatory milieu. Inspired by olfactory microenvironmental niches, we implemented a niche-targeted strategy. Our investigation defines the biological properties of ectodermal olfactory mesenchymal stem cells (OMSCs) and further elucidates their niche-targeted paracrine effectin vivoandin vitro. The findings demonstrate that OMSC-conditioned medium (OMSC-CM) delivered in fibrin hydrogel mediates potent SCI repair by concurrently protecting neurons, enhancing axonal regeneration, and suppressing destructive inflammation via IL-10 signaling. Critically, persistence of IL-10 signalingin vivois sustained both by direct supply from OMSC-CM and by OMSC-CM-induced activation of CD206⁺macrophages^IL-10. Successful neural circuit reconstruction with OMSC-CM depends on maximizing neuronal involvement in neural pathway formation. These findings may establish a special conceptual framework for developing regenerative medicine strategies in the future.

Stiffening of the extracellular matrix underlying the epithelial cells of the large intestine is associated with aging as well as many diseases. Yet the impact of the stiffened matrix on epithelial physiology remains poorly understood. A 2D and 3D microphysiological model of the large intestine was developed using a collagen scaffold with a physiologic or excessive stiffness (Young's moduli of 2.84 ± 0.85 kPa and 15.9 ± 0.73 kPa) by altering the collagen concentration within the substrate. Diffusion of a 10 and 40 kDa fluorescent dextran was significantly different between the physiologic and stiff scaffold (97.8 vs 79.8µm²s^-1[10 kDa] and 68.2 vs 56.8µm²s^-1[40 kDa], respectively). When primary human epithelial cells of the large intestine were grown as a 2D monolayer, cultures on the physiologic scaffold grew to a significantly higher density with more proliferative and fewer differentiated cells than cultures on the stiffened scaffold. Three-dimensional crypt arrays were also fabricated with the physiologic and stiff substrates, populated with cells, and a growth factor gradient applied. The cell density, proliferation, and height-to-width ratio was significantly greater for cells on the physiologic scaffold relative to that of cells on the stiffened scaffolds. Placement of a layer of intestinal fibroblasts below the epithelium on the crypt arrays did not mitigate the impact of the stiffened substrate. Bulk-RNA sequencing revealed 378 genes that were significantly upregulated and 385 genes significantly downregulated in the stiffened vs physiologic scaffolds. This work demonstrates that a molded collagen hydrogel can be used to mimic the biophysical characteristics of a stiffened intestinal stroma, recapitulating physiology observedin vivo. Thisin vitromodel of polarized crypts with a tunable underlying substrate will enable an improved understanding of intestinal epithelial cell morphology, stem cell maintenance and lineage allocation within a stiffened environment.

Replicating the complex mechanical and biological properties of native tissues remains a key challenge in three-dimensional bioprinting due to the limitations of single-nozzle systems. Here we present a multi-nozzle alternating bioprinting platform that addresses these problems by enabling precise control of mechanical and bioactive components' composition and distribution. By alternating cell-laden bioinks with mechanically reinforcing inks, our method enables precise spatial control for fabricating complex, anisotropic tissue architectures. A tri-layer printing strategy, using heart valve leaflets as a demonstrative model, was developed. In detail, gelatin methacryloyl bioinks, incorporating with porcine aortic valve interstitial cells and bioactive substances (e.g. basic fibroblast growth factor, polyaspartic acid, or chondroitin sulfate) to support cell function, are alternated with pluronic F-127 diacrylate mechanical reinforcement inks. This approach enhanced mechanical integrity of the constructs while supporting collagen, proteoglycan, and elastin production. Crucially, the constructs' mechanical robustness allowed direct cyclic mechanical stimulation during culture, further promoting tissue functional maturation and extracellular matrix remodeling.In vivo, the constructs showed excellent biocompatibility, with minimal calcification and favorable immune responses. This multi-material bioprinting platform enables the fabrication of tissue models that meet both structural and functional requirements, and can be adapted for a wide range of heterogeneous tissue and organ engineering applications, with the potential to significantly advance regenerative medicine.

A comprehensive understanding of the female reproductive system is essential for safeguarding fertility and preventing diseases related to women's health. Organoid/organ-on-a-chip (OOC), as a promising platform, could simulate complex physiological and pathological conditions, has revolutionized our understanding and management of female reproductive health. This technology is anticipated to advance the development of more effective assisted reproductive techniques, treatments and drug screening methods. This review focuses on various organoid/ OOC models designed to mimic the ovary, uterus, mammary gland, and vagina. Then, we introduce the current state of organoid/ OOC in female reproductive health and highlight how these models contribute to the study of female reproductive diseases. Additionally, we discuss the limitations of organoid/ OOC technology as well as its challenges and perspectives. Collectively, we believe that as organoid/ OOC technology continues to evolve, it holds great potential for transforming the diagnosis and treatment of female reproductive disorders, thereby enhancing women's overall health and well-being worldwide.

Diabetes, including all forms, remains a critical global health issue, affecting over 589 million adults and causing approximately 3.4 million deaths annually. Developing more relevantin vitromodels for pancreatic islet functions is crucial for advancing diabetes research and therapy. Microfluidic platforms enable precise control over experimental conditions, notably the mechanical cues within the tissue microenvironment, thereby offering a powerful tool for studying cell behavior under physiologically relevant conditions. In this study, we introduce an automated stimulation platform for single-islet glucose-stimulated insulin secretion, while insulin quantification remains off-chip. This platform incorporates an integrated micro-pump and automated fluid handling, obviating the need for external injection devices. Using both EndoC-βH5® spheroids and human donor islets, we demonstrate that the platform ensures high islet viability and functionality. This scalable and reproducible system represents a significant advancement in-depth studies of islet function, with broad applications for diabetes research and personalized treatment strategies.