Biofabrication最新文献

{"title":"Light-activated cartilage decellularised extracellular matrix hydrogels for engineering chondrogenic microenvironments with localised oxygen control.","authors":"Lin Li, Louis Jun Ye Ong, Khoon S Lim, Chamikara Liyanage, Yunkun Qu, Jingyi Wen, Caitlin Williams, Thomas G Molley, Roberto A Barrero, Shital Wakale, Ross Crawford, Kristopher A Kilian, Yi-Chin Toh, Indira Prasadam","doi":"10.1088/1758-5090/ae61f5","DOIUrl":"10.1088/1758-5090/ae61f5","url":null,"abstract":"<p><p>Cartilage tissue engineering requires biomaterials that can effectively maintain the tissue-specific functions of chondrocytes to enable the restoration of cartilage structure and function. Decellularised extracellular matrix (dECM)-derived hydrogels serve as tissue-specific biomaterials capable of preserving native biochemical cues and maintaining physiological chondrocyte phenotype in three-dimensional culture. However, their sol-gel transition relies heavily on collagen fibrillogenesis, a slow and poorly controllable process that limits mechanical tunability and suffers from inter-batch variability. Therefore, further efforts are required to functionalise cartilage dECM to achieve reproducible and controllable physicochemical properties. Here, we present a light-activated cartilage dECM hydrogel system based on ruthenium/sodium persulfate (Ru/SPS)-mediated dityrosine crosslinking, enabling rapid hydrogel formation under visible light irradiation while providing tunable mechanical properties and improved biological functionality. Comparison of the decellularisation protocols indicated that Triton X-100 combined with ammonium hydroxide efficiently eliminated residual DNA while preserving a substantial proportion of the native cartilage proteome. Pepsin-solubilised cartilage dECM hydrogels formed via dityrosine-based photo-crosslinking exhibited rapid gelation behaviour and superior mechanical characteristics compared to conventional thermally gelled dECM. The photo-crosslinked dECM hydrogels were cytocompatible, supported human bone marrow-derived mesenchymal stem cells (hBMSCs), and favoured cartilage-specific phenotypes, as demonstrated by the upregulation of chondrogenic genes, including<i>COL2A1</i>and<i>ACAN</i>, compared with gelatin methacrylate (GelMA) hydrogels. Importantly, this photo-crosslinking strategy overcomes the incompatibility between oxygen-sensitive redox-based photochemistry and hypoxic culture conditions, enabling the incorporation of oxygen-scavenging microcapsules to establish low-oxygen microenvironments. Under hypoxia, the cartilage dECM hydrogels promoted a more articular-like phenotype in hBMSC-derived chondrocytes, with transcriptomic features associated with TGF<i>-β</i>/SMAD2/3 and IGF-1/2-IGF-1R signalling. Collectively, these findings establish photo-crosslinked cartilage dECM hydrogels as a biomaterial platform with tunable mechanical properties and favourable biological functionality for cartilage tissue bioengineering and biomimetic<i>in vitro</i>cartilage models.</p>","PeriodicalId":8964,"journal":{"name":"Biofabrication","volume":" ","pages":""},"PeriodicalIF":8.0,"publicationDate":"2026-05-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147728236","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Spheroid assembly in microwells of defined geometry for quantitative assessment of aggregation kinetics and shape engineering.","authors":"Yuri M Efremov, Ekaterina Yu Makarova, Polina I Koteneva, Daniil O Golubchikov, Ruslan M Yanbarisov, Yuri V Vassilevski, Nastasia V Kosheleva, Peter S Timashev","doi":"10.1088/1758-5090/ae63f9","DOIUrl":"10.1088/1758-5090/ae63f9","url":null,"abstract":"<p><p>Three-dimensional (3D) cell spheroids are widely used as<i>in vitro</i>tissue models, yet quantitative understanding of their morphogenesis remains limited. We present an integrated experimental-computational framework to analyze, model, and modulate the compaction of cell aggregates in agarose microwells of defined geometries. Custom 3D-printed stamps produced circular, square, and triangular microwells of equal cross-sectional area. Time-lapse imaging combined with AI-based segmentation enabled tracking of spheroid morphology, with circularity and projected area serving as quantitative descriptors of compaction. The process followed predictable exponential kinetics, with mesenchymal (HDF) spheroids compacting faster than epithelial (ARPE-19) ones. Computational fluid dynamics (CFD) simulations modeled spheroid rounding as a visco-capillary-driven process, where the extracted visco-capillary velocity unified experimental and simulated dynamics. Mechanical measurements by atomic force microscopy and compression confirmed that differences in surface tension predominantly governed the observed kinetics. Pharmacological modulation of cytoskeletal tension revealed that inhibition of contractility markedly altered spheroid formation dynamics, enabling the generation of stable, non-spherical aggregates. Using this principle as a shape-engineering strategy, we produced aggregates with distinct geometries (brick-like, prismatic, and star-shaped), characterized by an increased surface-to-volume ratio compared to conventional spheroids. Limitations of the approach include the use of pharmacological cytoskeletal modulation and constraints in geometric fidelity arising from printing resolution, agarose casting, cell filling, and intrinsic smoothing of sharp features during cell aggregation. Collectively, this work establishes a geometry-controlled platform for quantitative analysis of spheroid formation and mechanical behavior, and provides a versatile framework for designing cell aggregates with defined shapes.</p>","PeriodicalId":8964,"journal":{"name":"Biofabrication","volume":" ","pages":""},"PeriodicalIF":8.0,"publicationDate":"2026-05-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147761093","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A 3D-printed osteochondral scaffold with a dual biomimetic design of spatially organized lotus-radial microchannels and bioinspired nano-mineral precursors for efficient osteochondral regeneration.","authors":"Qi Jiang, Yicong Wu, Ziyu Ding, Bowei Huang, Yuqing Gu, Xianzhu Zhang, Yuxuan Huang, Hongwei Ouyang, Shufang Zhang","doi":"10.1088/1758-5090/ae61f6","DOIUrl":"https://doi.org/10.1088/1758-5090/ae61f6","url":null,"abstract":"<p><p>Osteochondral defects present substantial clinical challenges due to the complex, multilayered structure and distinct physiological properties of cartilage and subchondral bone. Here, we report a three-dimensional (3D)-printed osteochondral scaffold featuring a dual biomimetic design that integrates vertically oriented microchannels with bioinspired nano-mineral precursors. Specifically, a multifunctional hierarchical construct was developed by incorporating ultrasmall (∼1 nm) polymer-induced liquid precursor-modified amorphous calcium phosphate (nCaP) into a GelMA-based matrix. Using digital light processing-based 3D printing, a biphasic scaffold with spatially defined architectures was fabricated, consisting of a pure GelMA upper layer featuring combined lotus-like and radial pore distributions to emulate the cartilage microenvironment, and a nCaP/GelMA lower layer with lotus-like pore architecture to support subchondral bone regeneration. Notably, in contrast to conventional inorganic fillers such as nanohydroxyapatite (nHAp), the incorporation of ultrasmall nCaP nanoclusters did not adversely affect photopolymerization behavior or printing fidelity, thereby enabling high-resolution fabrication. Beyond structural advantages, nCaP incorporation markedly enhanced the bioactivity of the scaffold. Compared with nHAp, nCaP significantly promoted the recruitment and osteogenic differentiation of endogenous bone marrow-derived mesenchymal stem cells, while also facilitating extracellular matrix deposition, mineralization, and angiogenesis. Transcriptomic analysis further indicated that these effects were associated with the upregulation of angiogenic factor EGFL6, suppression of inflammation-related TNFSF14/NF-<i>κ</i>B signaling, and activation of the PI3K-Akt pathway. Collectively, both<i>in vitro</i>and<i>in vivo</i>evaluations demonstrated that the nCaP/GelMA scaffold achieved improved tissue integration, restoration of hierarchical architecture, and enhanced mechanical performance compared with control groups. These findings underscore the potential of dual biomimetic scaffold design as an effective strategy for osteochondral regeneration.</p>","PeriodicalId":8964,"journal":{"name":"Biofabrication","volume":"18 2","pages":""},"PeriodicalIF":8.0,"publicationDate":"2026-05-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147833121","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Principle-based multiphysics simulation for 3D bioprinting systems: modelling inkjet, extrusion, and DLP processes.","authors":"Yunong Yuan, Ahmad-Fahmi Anwar-Fadzil, Hing Wai Chloe Choi, Tae-Joon Jeon, Yiqiao Hu, Jinhui Wu, Nezamoddin N Kachouie, Lifeng Kang","doi":"10.1088/1758-5090/ae6ad0","DOIUrl":"https://doi.org/10.1088/1758-5090/ae6ad0","url":null,"abstract":"<p><p>Among additive manufacturing (AM), 3D inkjet technology, materials extrusion (ME), and digital light processing (DLP), which are from dot and line to face printing, have been extensively investigated for biological and pharmaceutical applications. These techniques are valued for their ability to create customized complex drug laden devices and tissue engineering scaffolds. However, testing new bioinks or filament designs can be both expensive and time-consuming. To this end, numerical simulation offers a useful solution by reducing costs and saving time. Both machine learning and theory-based models can be used for simulation. Machine learning excels in handling complex data but faces challenges with data availability and overfitting, while theory-based models provide a more interpretable and data-efficient framework. This review explores how theory-based numerical simulation can be used to assess and optimize factors such as bioink printability, technique mechanism, printing parameters, and post-printing outcomes. By using simulation, key parameters can be understood and optimized without the need for physical experiments. The review highlights current models and discusses opportunities and challenges in using simulations to enhance the AM process, potentially advancing regenerative medicine and personalized treatments.</p>","PeriodicalId":8964,"journal":{"name":"Biofabrication","volume":" ","pages":""},"PeriodicalIF":8.0,"publicationDate":"2026-05-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147855776","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Development of ECM-inspired supramolecular cryogels with innate mineralization and compression-resistance for 3D culture of mini-bone trabeculae tissue analogs.","authors":"Shaoshuai Song, Wenxing Li, Ya Fang, Zhen Fang, Hong Zeng, Dandan Li, Youzhuan Xie, Junjie Niu, Jinwu Wang","doi":"10.1088/1758-5090/ae67a5","DOIUrl":"https://doi.org/10.1088/1758-5090/ae67a5","url":null,"abstract":"<p><p>Bone microtissue grafts mimicking skeletal features and organogenesis are an emerging strategy, different from the traditional tissue engineered bone grafts using the 3D cell encapsulation and the top cell seeding, and remain challenging in regenerative medicine and drug discovery, because the existing scaffold-free and microcarrier-based microtissue systems are difficult to manipulate the microtissue morphologies towards native bone microstructures limited by their mechanical weakness and the absence of interconnected inner cavities. Herein, we synthesized a supramolecular cryogel by an ice-templated freezing-polymerization process, acquiring a promising microcarrier resembling native bone trabecular morphology for 3D culture of trabecular bone microtissue grafts. In our strategy, a macromolecular chitosan monomer and two supramolecular monomers including glycinamide and phytic acid constituted the supramolecular cryogel, the modification using glycinamide and phytic acid components enables the cryogel microcarrier with porous cavities and compression-resistant abilities like the native trabecular bone tissues. Moreover, the mineralization of the cryogel microcarriers was also improved by the modification of phytic acid monomers, consequently strengthening osteogenic differentiation of bone-marrow-derived mesenchymal stem cells (BMSCs) and in vitro microtissue biomineralization. The in vivo results also revealed that a trabeculae-like bone microtissue forms on the cryogel microcarriers, with vessel invasion into inner cavities of the trabecular bone microtissues. After in situ implantation of the prepared trabecular bone microtissues, the bone regeneration characterized by raising bone mineral density and remodeling bone trabecular microstructures was observed on a rat femur condyle defect model. Last but not least, we also discovered the intestinal bacterial communities and compositions are closely related to the bone regeneration after implantation of the engineered bone microtissue grafts, which is the first evidence focusing on the intestinal microbiota response to the bone injury and bone regeneration events representing a feasible approach to bone regeneration examination. In brief, the supramolecular cryogels we developed in this study have been proved to be a promising microcarrier for the construction and 3D culture of trabecular bone microtissues, and this work offers a novelty insight into microtissue engineering and bone regeneration.</p>","PeriodicalId":8964,"journal":{"name":"Biofabrication","volume":" ","pages":""},"PeriodicalIF":8.0,"publicationDate":"2026-05-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147810921","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Bioprinting dynamic hydrazone hydrogels via light-mediated crosslinking.","authors":"Friederike Dehli, Forrest Hyde, Alexander Southan, Daniela F Duarte Campos","doi":"10.1088/1758-5090/ae67a6","DOIUrl":"https://doi.org/10.1088/1758-5090/ae67a6","url":null,"abstract":"<p><p>Viscoelastic hydrogels crosslinked by dynamic bonds hold great promise for mimicking the matrix dynamics of native tissues in cell culture and tissue engineering. Yet, their application in light-based bioprinting remains largely unexplored due to the incompatibility between reversible bond formation and photocrosslinking. This study addresses this key challenge by presenting a new class of photocrosslinkable, hydrazone-based bioinks developed from two modified polymers (Gel-A-DAAM and Gel-C-DAAM). These polymers are designed to enable reversible bond formation within hydrogel networks by attaching polymerizable groups to the polymer backbone via modular hydrazone conjugation chemistry. The resulting materials exhibit distinct mechanical properties depending on their hydrazide substituent, swelling medium, incubation temperature, and incubation time. Storage moduli of produced hydrogels vary between 0.08 - 1.2 kPa, which spans multiple scales of physiologically relevant tissue environments. The novel bioinks are suitable for droplet-based bioprinting followed by light-based crosslinking, and support cell spreading of human fibroblasts. Notably, the morphology of encapsulated cells varies with different hydrazide substituents, highlighting the potential of the developed bioink system to systematically investigate cell-matrix interactions. The combination of biological tunability and printability positions this system as a promising platform for fabricating next-generation tissue-mimetic constructs using advanced bioprinting technologies.</p>","PeriodicalId":8964,"journal":{"name":"Biofabrication","volume":" ","pages":""},"PeriodicalIF":8.0,"publicationDate":"2026-05-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147810944","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Development of a spatially defined 3D<i>in vitro</i>coculture construct modeling pancreatic cancer-associated cachexia.","authors":"Mitchell Kuss, Mena Asha Krishnan, Wonmi So, So-Youn Kim, Bin Duan","doi":"10.1088/1758-5090/ae5fda","DOIUrl":"10.1088/1758-5090/ae5fda","url":null,"abstract":"<p><p>Pancreatic cancer-associated cachexia is marked by adipose tissue wasting, thermogenic remodeling, and a state of hypermetabolism, yet robust preclinical models to study these phenomena are lacking. In this study, we present a spatially defined three-dimensional (3D) core-shell microcuboid coculture platform designed to investigate the interaction between adipocytes and pancreatic cancers. This innovative system consists of differentiated white adipocytes at the core, surrounded by pancreatic ductal adenocarcinoma (PDAC) cells embedded in 3D-printed microcuboids, arranged concentrically within a collagen coculture matrix construct. Within this framework, we observed significant enhancement of adipocyte lipolysis and browning, as evidenced by BODIPY dye-tracked lipid migration, sustained glycerol release, and progressive expression of extracellular UCP1 or the mitochondrial brown fat uncoupling protein 1, particularly pronounced in cocultures involving aggressive pancreatic cancer cell lines. The integrity of the core-shell architecture persisted for up to 21 d but progressively disintegrated under the influence of the cancer cells marked by cancer cell invasion into the adipocyte regions. Gene profiling revealed a downregulation of adipogenic markers, such as<i>Pparg, Plin1</i>, and<i>Lipe</i>, alongside an increase in<i>Ucp1</i>transcripts, suggesting a metabolic shift from lipid storage to utilization and thermogenic activation. In contrast to existing 3D engineered systems, our platform offers enhanced long-term viability, controlled compartmentalization, mechanical tunability, and high spatiotemporal resolution. It effectively recapitulates the dynamic interplay between cancer and adipose cells, along with the catabolic characteristics of PDAC-associated cachexia, serving as a scalable<i>in vitro</i>tool for mechanistic investigations, and for testing potential anti-cachexia interventions, filling the gap between simplistic<i>in vitro</i>assays and complex animal models.</p>","PeriodicalId":8964,"journal":{"name":"Biofabrication","volume":" ","pages":""},"PeriodicalIF":8.0,"publicationDate":"2026-04-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147687761","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Micro-comb 3D printing: rapid fabrication of tissue-guiding substrates using micro-embossed nozzles.","authors":"Sarkhan Butdayev, Stefan Leone, Shayla Nikzad, Janko Kajtez, Katrine Bech Lauritzen, Moises Di Sante, Francesco S Pasqualini, Kirstine Calloe, Rodolphe Marie, Stephan S Keller, Anne Z Eriksen, Johan U Lind","doi":"10.1088/1758-5090/ae5fd9","DOIUrl":"10.1088/1758-5090/ae5fd9","url":null,"abstract":"<p><p>To replicate the function of native tissue in cell cultures, one must reproduce the structure of the native tissue. This can be achieved using tissue-guiding architectures with cell-scale dimensions, typically ranging from single to tens of microns. However, this spatial resolution exceeds the capabilities of many common fabrication methods, including extrusion-based 3D printing. Indeed, although increasingly popular in bioengineering, extrusion-based 3D printing is not only limited by the properties of the print materials, but also by the inherent trade-off that smaller features require smaller nozzles. This, in turn, results in more toolpaths and longer build times. To overcome this limitation, we introduce nozzles with micro-scale structures at their orifice, fabricated through straightforward hot embossing of commercial polypropylene nozzles. This approach enables microstructure printing using large (⩾0.4 mm inner diameter) nozzles. Specifically, we demonstrate rapid printing of microstructured soft substrates, capable of guiding skeletal and cardiac muscle cell cultures into physiomimetic, anisotropic tissues for electrophysiological assays and drug studies. Furthermore, we show that axonal growth in neuronal tissue cultures can also be directed. Thus, our approach may serve as a scalable and easily accessible method for fabricating human cell cultures and tissue models with enhanced physiological relevance.</p>","PeriodicalId":8964,"journal":{"name":"Biofabrication","volume":" ","pages":""},"PeriodicalIF":8.0,"publicationDate":"2026-04-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147687782","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Extruded droplet-on-demand (X-DoD) bioprinting for controlled iPSC-based functional cortical network formation.","authors":"Elisabeth Riska, Roni Cohen, Lior Perry-Tomer, Yahel Shechter, Eric Silberman, Assaf Shapira, Tal Dvir","doi":"10.1088/1758-5090/ae647c","DOIUrl":"https://doi.org/10.1088/1758-5090/ae647c","url":null,"abstract":"<p><p>Engineered three-dimensional (3D) neural constructs hold significant promise for repairing neural tissue damage and recapitulating the human brain in vitro for disease modeling and drug screening applications. However, most current 3D neural models, including freestanding organoids and dense bioprinted neural constructs, lack the architectural and functional organization required to emulate the cerebral cortex, which comprises grey matter regions rich in neuronal cell bodies and white matter tracts formed by long-range axonal projections. This architectural mismatch limits the models' ability to support functional connectivity analysis, predict in vivo behavior, and achieve effective integration with host tissue. In this study, we present an extruded droplet-on-demand (X-DoD) bioprinting technique that enables deterministic spatial patterning of droplets containing human induced pluripotent stem cells (iPSCs) encapsulated within an extracellular matrix (ECM)-based hydrogel and embedded in a permissive, low-concentration hydrogel bulk that supports diffusion. Using a 5×5 droplet array pattern, we demonstrate that after 30 days of differentiation into cortical neurons and formation of 3D neuronal networks, the micron-scale, cell-body-dense droplets (microtissues) remain localized at their initial droplet sites and are interconnected by millimeter-scale neurite projections. This defined grey-white matter-like organization enables functional analysis via calcium imaging and seamless integration with custom electronic devices for advanced neurophysiological interrogation. Calcium imaging and electrical recordings revealed temporally preserved, propagating network activity, with network excitability dynamically modulated by treatment with GABA antagonist bicuculline. Altogether, the X-DoD bioprinting platform offers a powerful and adaptable approach for engineering spatially organized 3D neural networks with tunable connectivity, providing a robust tool for studying brain function, disease modeling, and future therapeutic applications.</p>","PeriodicalId":8964,"journal":{"name":"Biofabrication","volume":" ","pages":""},"PeriodicalIF":8.0,"publicationDate":"2026-04-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147761103","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Organobodies: a robust and size-controllable system for generating scalable hiPSC-derived liver organoids for drug toxicity screening.","authors":"Mostafa Kiamehr, Stefano Manzini, Burak Toprakhisar, Rodrigo F Madeiro da Costa, Guillem García-Llorens, Birhanu Belay, Mustapha Najimi, José V Castell, Wolfgang Moritz, Giulia Chiesa, Katriina Aalto-Setälä, Catherine Verfaillie","doi":"10.1088/1758-5090/ae5fd8","DOIUrl":"10.1088/1758-5090/ae5fd8","url":null,"abstract":"<p><p>Human hepatic organoids derived from pluripotent or adult stem cells offer powerful platforms for disease modeling and drug discovery. However, developing robust and scalable organoids capable of sustaining long-term functionality remains challenging. Here, we developed a novel, semi-defined approach using a self-assembling peptide and collagen I to create highly uniform human induced pluripotent stem cell-derived hepatic organoids in droplet format, which we term hepatic organobodies (OBs). This method enabled rapid, reproducible production of threedimensional (3D) liver tissues, which remained structurally, metabolically, and functionally stable for several weeks. OBs adopted hallmark hepatic morphology and expressed key hepatocyte genes, several at levels approaching freshly isolated primary human hepatocytes (PHHs). OBs secreted substantially higher albumin and A1AT compared with parallel two dimensional cultures, and transcriptomic profiling revealed marked enhancement of hepatic maturation, including elevated expression of<i>CYP3A4, CYP2C9</i>, and<i>CYP1A2</i>, and enrichment of PPAR signaling and fatty acid<i>β</i>-oxidation pathways. Additionally, OBs exhibited drug metabolizing activity comparable to classical Matrigel-based organoids and demonstrated CYP3A4 and CYP2C9 activities comparable to the 'gold standard' 3D PHH microtissues. Critically, OBs accurately predicted hepatotoxicity of more than 10 reference compounds, outperforming HepG2 cells and matching PHH-based benchmarks. Overall, we present OBs, a novel, and scalable 3D liver model that delivers advanced maturation and robust metabolic function. This platform offers a powerful and reproducible alternative to existing organoid systems as it avoids animal-derived, undefined matrices such as Matrigel, requires no specialized equipment, and relies on rapid self-curation of the hydrogel triggered by physiological salt concentrations, making the process fast, reproducible, broadly accessible, and scalable.</p>","PeriodicalId":8964,"journal":{"name":"Biofabrication","volume":" ","pages":""},"PeriodicalIF":8.0,"publicationDate":"2026-04-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147687886","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}