Biofabrication最新文献

{"title":"Sequential stenciling to reconstitute 2D microtissues for multicellular and synthetic signaling architectures.","authors":"Kai Hirzel, Jasmin Čić, Stella Asmanidou, Nadja Schmohl, Roland E Kontermann, Satoshi Toda, Monilola A Olayioye, Andrew G Clark, Michael Heymann","doi":"10.1088/1758-5090/ae9807","DOIUrl":"10.1088/1758-5090/ae9807","url":null,"abstract":"<p><p>In mammalian organisms, native tissue function depends on precise spatial organization down to the cellular level. Reconstituting tissue architectures in 2D<i>in</i><i>vivo</i>platforms can provide a means to study direct and indirect cell-cell interactions in a variety of tissue contexts while remaining compatible with high-throughput assays and high-resolution live imaging. We combine cost-effective stereolithography leveraging 3D printing with replica molding to stencil spatially defined, multicellular culture systems with sub-millimeter resolution onto planar substrates. The system is designed for ease of use, requires no complex fabrication setups and scales readily to 96-well plates. Sequential stencil application and removal under a biosafety cabinet enables controlled positioning of multiple cell types and supports the maturation of tissue assemblies. We demonstrate the utility of this stencil-based patterning strategy in three applications. First, we employ a combination of two circular stencils to recreate a structural feature characteristic of the tumor microenvironment of solid tumors: the encapsulation of colorectal cancer cells by cancer-associated fibroblast (CAF) cells. Resulting cell patterns recapitulate key aspects of native tissue dynamics of the densely packed tumor tissues, in which CAF cells actively compress the cancer cells and confer targeted therapy resistance. Second, we probe an engineered synNotch-based signaling system in patterned cell patches that mimics morphogen gradient formation, where GFP-secreting sender cells generate a ligand-dependent gradient. Third, we recapitulate the characteristic crypt-villus architecture of the mammalian intestine by patterning intestinal organoids within a stencil-restricted crypt region and allowing differentiating cells to collectively migrate along a designed villus axis. The presented strategy allows for rebuilding multicellular tissue architectures<i>in</i><i>vitro</i>with biologically relevant spatial precision for high-throughput drug screenings and dissection of tissue-specific cellular interactions.</p>","PeriodicalId":8964,"journal":{"name":"Biofabrication","volume":" ","pages":""},"PeriodicalIF":8.2,"publicationDate":"2026-09-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148705253","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 high-density microelectrode array integrated microfluidic platform for quantitative investigation of functional connectivity in interconnected human dopaminergic neurospheroids.","authors":"Chiara Ausilio, Annachiara Scalzone, Valentina Mollo, Paolo Antonio Netti","doi":"10.1088/1758-5090/aea7de","DOIUrl":"10.1088/1758-5090/aea7de","url":null,"abstract":"<p><p>The quantitative investigation of functional connectivity across spatially organized neuronal networks remains a major challenge in human three-dimensional (3D)<i>in vitro</i>models. Although neurospheroids, brain organoids, and microfluidic systems enable controlled structural organization and guided neurite growth, their integration with high-resolution electrophysiological interfaces remains limited, often requiring post hoc adaptations that compromise recording stability and long-term reproducibility. Here, we present a polydimethylsiloxane (PDMS) microfluidic platform natively engineered for seamless integration with high-density microelectrode arrays (HD-MEAs), enabling the controlled interconnection and long-term electrophysiological interrogation of human neurospheroids within a defined geometry. The platform supports reproducible axonal growth between spatially separated spheroids while preserving full compatibility with HD- microelectrode arrays (MEA) recording requirements and electrical stability. Structural characterization revealed the formation of a stable inter-spheroid axonal fascicle enriched in neuronal and synapse-associated markers, supported by ultrastructural analyzes across multiple spatial scales. Long-term electrophysiological recordings revealed progressive electrophysiological maturation-associated changes, characterized by increased mean firing rate, sustained and spatially confined neuronal activity within each spheroid, and changes in burst dynamics. Quantitative cross-correlation analysis identified lag-consistent interaction patterns compatible with both local (intra-spheroid) and long-range (inter-spheroid) putative functional connectivity, enabling the distinction between local and distributed network dynamics within the modular system. By combining controlled 3D architecture, stable HD-MEA integration, and quantitative electrophysiological readouts, this platform establishes a robust, reproducible and human-relevant framework for investigating activity patterns compatible with functional connectivity dynamics in interconnected neuronal assemblies, with potential applications in disease modeling and connectopathies-related studies.</p>","PeriodicalId":8964,"journal":{"name":"Biofabrication","volume":" ","pages":""},"PeriodicalIF":8.2,"publicationDate":"2026-09-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148975252","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":"CORRIGENDUM: Biofabrication of 3D bioprinted and organ-on-chip blood-brain barrier models using hCMEC/D3 for intranasal delivery of central nervous system therapeutics (2026<i>Biofabrication</i> 18 012005).","authors":"Chun Yuen Jerry Wong, Thara Pannadewi, Tanisha Tabassum Sayka Khan, Hui Xin Ong, Daniela Traini","doi":"10.1088/1758-5090/aea9ce","DOIUrl":"https://doi.org/10.1088/1758-5090/aea9ce","url":null,"abstract":"","PeriodicalId":8964,"journal":{"name":"Biofabrication","volume":"18 4","pages":""},"PeriodicalIF":8.2,"publicationDate":"2026-09-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149069308","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":"Patient-specific 3D-printed mold system for superior semicircular canal third-window repair: design, fabrication, and first-in-human feasibility.","authors":"Ahmad Alshehri, Sara Alhazani, Mohammed Abdul Fasi","doi":"10.1088/1758-5090/aea59f","DOIUrl":"10.1088/1758-5090/aea59f","url":null,"abstract":"<p><p>Superior semicircular canal (SSC) dehiscence is a pathological defect in the bony labyrinth of the inner ear that produces auditory and vestibular symptoms due to abnormal third-window mechanics. Conventional surgical methods, including canal plugging and resurfacing, may improve symptoms but can be associated with vestibular or auditory risks. This study reports the design, fabrication, technical verification, and first-in-human feasibility of a patient-specific, 3D-printed mold system for anatomical repair of the SSC. Using high-resolution computed tomography, three-dimensional reconstruction, and computer-aided design, a patient-specific mold conforming to the external SSC roof was fabricated. The mold enables extracorporeal shaping of polymethyl methacrylate (PMMA) bone cement outside the patient to create a patient-specific implant for closure of the pathological third-window defect with precise anatomical alignment. Technical verification using a 3D-printed temporal bone model demonstrated anatomical fit and defect coverage before clinical use. The system was applied in a first-in-human clinical feasibility case, with early postoperative symptom improvement/resolution and no clinically evident new vestibular or auditory deterioration during early assessment. Preoperative and postoperative audiometric and vestibular test results, including general VEMP and vHIT data, are reported to support transparent interpretation of the early clinical outcome. This first-in-human feasibility report suggests that patient-specific additive manufacturing may support precision otologic reconstruction, while further clinical studies are required to establish broader safety, durability, reproducibility, and clinical effectiveness.</p>","PeriodicalId":8964,"journal":{"name":"Biofabrication","volume":" ","pages":""},"PeriodicalIF":8.2,"publicationDate":"2026-09-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148928804","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":"Structural reconstruction of spinal cord white matter using a biomimetic zoned fiber-hydrogel scaffold.","authors":"Ting-Ting Zheng, Ying Han, Hailin Ma, Jixiang Zhu, Wensheng Lin, Jia-Hui Sun, Jian-Long Zou","doi":"10.1088/1758-5090/ae8a81","DOIUrl":"10.1088/1758-5090/ae8a81","url":null,"abstract":"<p><p>Following spinal cord injury (SCI), the zonal architecture of gray and white matter remains challenging to reconstruct. The present study explores a strategy for reconstructing this zonal structure via the integration of oriented fibers with hydrogels. The SFs were rationally arranged to conform to the morphological features of the white matter. The incorporation of the hydrogel, in turn, filled the gray matter regions and immobilized the spatial distribution of the fibers. This gray/white matter zoned scaffold (GWZS) effectively mimics the inherent characteristics of native spinal cord gray and white matter. When dorsal root ganglion neurons were 3D cultured within this GWZS, gray/white matter-analogous morphogenesis was recapitulated. Specifically, in the gray matter region, neuronal clusters and nerve fiber network-like architectures were formed; in contrast, in the white matter region, oriented axonal bundles were formed.<i>In vivo</i>transplantation of GWZS effectively facilitated the reconstruction of white matter in the spinal cord; however, endogenous neuroregeneration remained deficient in the gray matter region. Overall, this study not only establishes a novel preparation method for oriented fiber-hydrogel composite bionic spinal cord scaffolds but also provides a viable strategy for the reconstruction of white matter architecture following SCI.</p>","PeriodicalId":8964,"journal":{"name":"Biofabrication","volume":" ","pages":""},"PeriodicalIF":8.2,"publicationDate":"2026-09-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148444290","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 provisional and evolving framework for standardizing cartilage organoid construction: toward consistency, reproducibility, and clinical translation.","authors":"Hefei Zhang, Shengdong Yang, Junyou Li, Xinjie Wu, Wei Sun, Rongjie Feng, Fuqiang Gao, Qingyu Zhang","doi":"10.1088/1758-5090/ae9ef4","DOIUrl":"10.1088/1758-5090/ae9ef4","url":null,"abstract":"<p><p>Cartilage organoids (COs) have emerged as a promising three-dimensional platform for modeling cartilage development, degeneration, and repair, yet their broader application remains constrained by substantial variability in cell sources, construction strategies, culture conditions, and evaluation criteria. This manuscript addresses the critical need for standardization in CO research and aims to establish a structured framework for their classification, construction, application, and quality assessment. We systematically summarize the defining features of COs and distinguish them from related cartilage models, then review major cell sources, including embryonic stem cells, induced pluripotent stem cells, mesenchymal stem cells, and periosteum-derived cells, together with scaffold-free and scaffold-assisted fabrication strategies, stepwise induction protocols, bioprinting approaches, bioreactor systems, and key microenvironmental regulators such as hypoxia, mechanical stimulation, and multicellular interactions. Based on this, we propose a literature-informed conceptual framework termed COSS (Cartilage Organoid Standardization System). COSS comprises a three-tier standardization framework encompassing input control, process standardization, and output evaluation. It further defines minimum reporting items and a tiered evaluation framework for disease modeling, drug screening, and regenerative repair, with the aim of harmonizing terminology, reporting requirements, quality attributes, and priorities for future validation. This framework cannot yet be regarded as an experimentally validated production standard or a regulatory-endorsed system. Further work is required to refine the framework and establish reliable quantitative thresholds and specific numerical criteria. Collectively, this work provides an application-oriented roadmap for improving reproducibility, comparability, and translational relevance in CO studies. These recommendations could serve as an initial framework for multicenter benchmarking and for supporting future determination of reproducibility for mechanistic studies, preclinical evaluation, or repair-oriented applications.</p>","PeriodicalId":8964,"journal":{"name":"Biofabrication","volume":" ","pages":""},"PeriodicalIF":8.2,"publicationDate":"2026-09-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148825182","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":"Cell sheet engineering at an inflection point: from pioneering techniques to commercialized products.","authors":"Alireza Shahin-Shamsabadi","doi":"10.1088/1758-5090/aea1bf","DOIUrl":"10.1088/1758-5090/aea1bf","url":null,"abstract":"<p><p>Cell sheet engineering (CSE), a scaffold- and biomaterial-free biofabrication technique, preserving native extracellular matrix and cell-cell junctions, is re-emerging as a powerful therapeutic modality. With over 70 registered clinical trials worldwide, the technology has demonstrated clinical validity by providing preserved tissue architecture. Recent evolutions such as those in anchored cell sheet engineering, mark an inflection point, enabling formation of more relevant form factors with higher levels of phenotypical and microstructural maturity with improved structural and functional competence, expanding CSE's addressable scope. Regulatory momentum is growing, with the gap between current cell-based release testing and tissue-level function emerging as the central regulatory challenge for living tissue products. Simultaneously, automation is transforming CSE's cost structure, with closed-loop systems potentially reducing manufacturing costs by ∼30% from current facility- and personnel-dominated models. Commercial precedents demonstrate that biologically complex constructs can achieve reimbursement and clinical integration when aligned with procedural workflows and measurable outcomes. This perspective examines how CSE represents a high-value platform for a strategically significant and expandable subset of regenerative medicine, positioned to serve a substantial market segment. Future advances could expand its applications to larger multi-tissues constructs, where recreating native tissue architecture provides decisive therapeutic advantage.</p>","PeriodicalId":8964,"journal":{"name":"Biofabrication","volume":" ","pages":""},"PeriodicalIF":8.2,"publicationDate":"2026-09-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148879002","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":"3D bioprinting of stromal vascular fraction enriched with MXenes for enhanced intestinal regeneration.","authors":"Giordano Perini, Antonio Minopoli, Giada Di Giulio, Margherita Montescagli, Alberto Augello, Valeria Ferrara, Davide Evangelista, Matteo Marras, Giulia Artemi, Anna Amelia Caretto, Stefano Gentileschi, Dania Nachira, Dario Zambrano, Sana Munir, Valerio Pontecorvi, Cristiano Spada, Loredana Gualtieri, Andreas Rosenkranz, Valentina Palmieri, Ivo Boskoski, Marco De Spirito, Massimiliano Papi","doi":"10.1088/1758-5090/ae9b71","DOIUrl":"10.1088/1758-5090/ae9b71","url":null,"abstract":"<p><p>Intestinal disorders such as inflammatory bowel diseases and gastrointestinal fistulae are marked by chronic inflammation, barrier dysfunction, and impaired repair, conditions insufficiently addressed by current treatments. Regenerative strategies able to restore epithelial integrity and support stromal and vascular remodeling are therefore highly needed. The stromal vascular fraction (SVF), a heterogeneous and autologous cell population from adipose tissue, offers angiogenic, immunomodulatory, and regenerative properties, while three-dimensional (3D) bioprinting enables its embedding in hydrogels to enhance survival and activity. MXenes (MX), a novel class of two-dimensional materials, can help to add further advantages through their bioactive, antioxidant, and pro-angiogenic features. In this contribution, we engineered 3D bioprinted constructs composed of SVF embedded in GelMA functionalized with MX. After confirming the biocompatibility of MX on multiple cell types, we demonstrated that SVF MX constructs notably promoted wound closure, endothelial tube formation, and epithelial proliferation, while maintaining stable, non-toxic ROS levels and demonstrating excellent cytocompatibility. These findings highlight MX-enriched SVF constructs as an innovative platform for next-generation regenerative therapies, combining the autologous, low-immunogenic profile of SVF with the multifunctional properties of MX to address complex intestinal disorders.</p>","PeriodicalId":8964,"journal":{"name":"Biofabrication","volume":" ","pages":""},"PeriodicalIF":8.2,"publicationDate":"2026-09-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148787845","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":"Inkjet biofabrication of electroactive rGO/growth-factor micropatterns on electrospun PLCL for guided neural cell responses.","authors":"Haoyu Wang, Jin Tian, Yanshen Yang, Jiachen Pang, Xinci Duan, Feng Xu, Xiaqing Zhou","doi":"10.1088/1758-5090/ae9f98","DOIUrl":"10.1088/1758-5090/ae9f98","url":null,"abstract":"<p><p>Biofabrication of neural interfaces requires spatial control over conductive, biochemical, and topographical cues at the cell-contacting surface. Here, we report and validate an<i>in vitro</i>biofabricated screening platform based on electrospun poly(lactide-co-<i>ϵ</i>-caprolactone) (PLCL), in which graphene oxide (GO)/growth-factor precursor inks are inkjet-printed and reduced with ascorbic acid to form reduced graphene oxide (rGO)/growth-factor micropatterns. The platform was used to screen printing-layer number, nozzle configuration, and electric-field strength by integrating band-width fidelity, continuity, overspray, and apparent conductivity with neural-cell readouts. A 4-layer double-nozzle condition produced a core printed-band width of 204.8 µm, close to the designed 200 µm width, with 94.5% continuity and 0.50% overspray, while providing an apparent conductivity of 0.432 ± 0.038 S cm^-1, approximately 78.8% of the 10-layer condition. The same workflow localized nerve growth factor or neuregulin-1 at the basal cell-material interface. Fluorescence-assisted imaging verified post-reduction pattern retention, while ELISA quantified immunoreactive recovery and cumulative release. Across the tested models, the printed interface and stimulation conditions were associated with changes in neural cell behavior. In PC-12 cultures, 150 mV cm^-1direct-current stimulation was associated with greater<i>β</i>III-tubulin-positive neurite-like outgrowth and alignment than W/O ES or 300 mV cm^-1, reaching 19 060 ± 2 417 µm total neurite length per field at day 21. Dorsal root ganglion neurons extended aligned TUJ1-positive neurites along the printed tracks, and S16 Schwann cells expanded on neuregulin-1/rGO-patterned substrates under the same stimulation condition. RT-qPCR and western blotting showed higher neuronal regeneration-associated and Schwann-cell-associated marker levels under 150 mV cm^-1than under unstimulated or higher-field conditions. These results establish an<i>in vitro</i>screening platform for evaluating printing parameters and defined topographical, electrical, and biochemical cues within the tested neural cell models.</p>","PeriodicalId":8964,"journal":{"name":"Biofabrication","volume":" ","pages":""},"PeriodicalIF":8.2,"publicationDate":"2026-09-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148839095","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 low cell density model for examining astrocyte reaction to 3D suspension using AVIFJ.","authors":"Austin Smith, Yu Song, Wei Sun, Feng Lin","doi":"10.1088/1758-5090/ae9401","DOIUrl":"10.1088/1758-5090/ae9401","url":null,"abstract":"<p><p>Alternating viscous and inertial force jetting (AVIFJ) was used to print single-cells and position small numbers of cells precisely. The effect of nozzle size on single-cell encapsulation and droplet spacing ability was tested using astrocytes. Astrocytes were patterned in Matrigel and examined for 7 d after printing to assess viability and their response to being suspended in 3D at low cell density. The results showed AVIFJ had a single-cell printing efficiency around 30% using nozzles with outer diameters (ODs) of 63<i>μ</i>m and 96<i>μ</i>m at a 300 000 cells ml^-1concentration, and 30% for 143<i>µ</i>m and 195<i>µ</i>m OD nozzles at a 50 000 cells ml^-1concentration. Spacing using a 63<i>μ</i>m nozzle tests printed droplets with a minimum center-to-center spacing of 199.53 ± 2.52<i>μ</i>m on a culture well and cell spacing of 122.23 ± 21.69<i>μ</i>m in 3D. Astrocyte viability was 93.60 ± .27% when printed into Matrigel. RNA sequence results showed astrocytes suspended Matrigel upregulated IGFBP3 compared to 2D controls. Results also showed that astrocytes suspended in 3D that contact the culture well react differently than those completely suspended over 4 d. Overall, this study showed the viability of AVIFJ printing for low cell number experiments and differences in astrocyte behavior in 3D suspension compared to 2D.</p>","PeriodicalId":8964,"journal":{"name":"Biofabrication","volume":" ","pages":""},"PeriodicalIF":8.2,"publicationDate":"2026-09-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148668354","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}