Cell sheet engineering (CSE), a scaffold- and biomaterial-free biofabrication technique, preserving native extracellular matrix (ECM) 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 (ACSE), 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.
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.
Hemorrhage remains one of the leading causes of trauma-related mortality worldwide and poses a major challenge in surgery, emergency medicine, and battlefield rescue. Conventional hemostatic materials, such as gauze and gelatin sponges, mainly rely on passive blood absorption and compression, which often exhibit limited efficacy in irregular wounds and severe bleeding environments. In recent years, advances in biomaterials and tissue engineering have promoted the development of hemostatic sponges with enhanced absorption capacity, bioactivity, tissue adhesion, antibacterial performance, and wound healing ability. Compared with other hemostatic platforms, sponge-based systems offer unique advantages derived from their interconnected porous architectures, enabling rapid blood transport, coagulation component enrichment, mechanical wound filling, and structural adaptation in complex bleeding scenarios. In this review, we discuss the development of hemostatic sponges from the integrated perspectives of material systems, biofabrication strategies, structural regulation, and biomedical applications. Different classes of materials, including polysaccharides, proteins, and synthetic polymers, are systematically discussed with emphasis on their intrinsic hemostatic mechanisms, functional modifications, and design limitations. The relationship between sponge structures and hemostatic performance, including pore architecture, interconnected channels, shape-memory behavior, and surface functionality, is highlighted. Representative fabrication strategies and multifunctional applications, including antibacterial activity, wound healing, and non-compressible hemorrhage control, are summarized. Finally, current challenges and future perspectives regarding intelligent design, standardized evaluation, and clinical translation of next-generation hemostatic sponges are discussed. This review provides design principles and translational insights for developing advanced hemostatic sponge systems.
In mammalian organisms, native tissue function depends on precise spatial organization down to the cellular level. Reconstituting tissue architectures in 2Din vitroplatforms 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 cells. Resulting cell patternings recapitulate key aspects of native tissue dynamics of the densely packed tumor tissues, in which cancer-associated fibroblast 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 architecturesin vitrowith biologically relevant spatial precision for high-throughput drug screenings and dissection of tissue-specific cellular interactions.
Organoids derived from human pluripotent stem cells (PSCs) have emerged as powerfulin vitromodels for studying development, disease, and therapeutic responses, yet their lack of functional vasculature limits growth, maturation, and physiological relevance. Early vascularization strategies relied on human umbilical vein endothelial cells (ECs), which lack organ-specific identity and introduce donor variability. The field is now undergoing a paradigm shift toward PSC-derived vasculature, which offers patient-specific, and developmentally stage-matched endothelium with PSC-derived organoids. This review summarizes current strategies for organoid vascularization, with emphasis on both human PSC-derived 2D ECs and 3D blood vessels. Approaches relying on co-aggregation of differentiated ECs with organ-specific populations or external endothelial coating of pre-formed organoids. These improved survival and functional maturation but remain limited in spatial organization and perfusability. The advances have incorporated pre-formed vascular spheroids and iPSC-derived blood vessel organoids, which can be respectively fused with lineage-specific organoids to generate vascularized assembloids to enhance vascular architecture and tissue maturation. This review further highlights engineering the microenvironment to promote the formation of vascular niche, such as hypoxia modulation, transcriptional regulation, signaling transduction, and extracellular matrix engineering. In addition, we discuss the current limitations as well as future directions of vascularized organoids, including the unmet need for developing tissue-specific ECs, improved engraftment following transplantation, and organ-on-a-chip platforms. Collectively, integrating iPSC-derived vasculature within organoids provides a central framework toward physiologically relevant, perfusable tissues and expands the translational utility of organoid technologies for disease modeling and therapeutic development.
Lipid nanoparticles (LNPs) are widely used in nanomedicine, yet their energy-efficient manufacture with precise control over particle uniformity remains challenging. Here we report a gravity-driven microfluidic platform that integrates acoustically actuated sharp-tip mixing to enable high-throughput LNPs synthesis without the need for active pumps. By optimizing microchannel geometry, actuation conditions, and the spatial positioning of the mixing vortex, we achieve stable gravity-driven flow and complete mixing at total flow rates (TFRs) of up to 5 ml min-1. The optimized system produces LNPs with tunable sizes below 60 nm, polydispersity <0.3, and high production yields. We further show that nanoparticle uniformity is governed not only by mixing strength but also by where mixing and nucleation occur within the microchannel. These findings establish gravity-driven sharp-tip microfluidics as a scalable and energy-efficient strategy for controlled nanoparticle manufacturing, offering a practical route toward greener nanomedicine production.
Three-dimensional cell culture using microcarriers is an effective strategy for scalable cell expansion. However, conventional enzymatic detachment can compromise cell viability, surface proteins, and native signaling. We report viscoelasticity-tunable hyaluronic acid (HA)-gelatin microspheres as microcarriers, engineered with a thermoresponsive polymer coating to enhance cell attachment and enable gentle harvesting. Gelatin-only (GLA), gelatin-HA (G-HA), and gelatin-HA-L-lysine (G-HA-L) microspheres were fabricated. HA incorporation and lysine functionalization were used to tune microsphere mechanics and interfacial stability. Frequency-sweep rheology revealed that HA-containing formulations exhibited higher elastic dominance (G' > G″) and a broader, more stable viscoelastic response than gelatin-only and a commercial gelatin microcarrier benchmark, with G-HA-L showing the most favorable balance of stiffness and damping (highest G'/ G″ across the tested window). The microspheres were subsequently coated with poly (N-isopropylacrylamide-co-acrylic acid) (P(NIPAM-AAc)), producing a temperature-responsive interface. Importantly, the thermoresponsive coating enhanced early cell attachment, particularly on G-HA-L (reaching ∼70% within 4 h and approaching ∼90% by 24 h), outperforming both coated G-HA and commercial microcarriers. For harvesting, low-temperature conditioning markedly improved cell release and recovery compared to trypsin-only controls, consistent with temperature-triggered polymer swelling, which facilitated detachment. Collectively, these results demonstrate that coupling viscoelastic microcarrier design with thermoresponsive surface engineering provides a promising platform for efficient cell growth and gentle, process-friendly harvesting, with potential applications in tissue engineering and regenerative medicine.
For healing full-thickness skin defects caused by severe injury, skin substitute grafts with structural and compositional biomimicry are critical. Vascularized spheroids, as highly active biological building blocks, can be applied in biomanufacturing to fabricate functional repair constructs. However, current methods of biofabricating endothelialized spheroids still face multiple technical limitations, including low throughput, poor uniformity, and insufficient controllability and observability. Furthermore, there is inadequate control over the spatial arrangement of the spheroids during vascularized construct biomanufacturing. We proposed a synergistic, shape-controlled spheroid-hydrogel biomanufacturing method to address these issues, and developed a highly integrated, one-stop bioprinting platform for spheroids (OBPS) with real-time monitoring. We focused on key biomanufacturing stages, spheroid fabrication, bioprinting, and in vivo validation, to establish an integrated technical framework providing for "fabrication-culture-assembly-application." Using the OBPS, we fabricated endothelialized spheroids by co-culturing human fibroblasts and human umbilical vein endothelial cells, printed fibrinogen-supplemented gelatin methacryloyl hydrogel, and bioprinted spheroids and hydrogels in situ onto full-thickness skin defects in nude mice. Animal experiments show that shape-controlled endothelialized spheroids significantly accelerated wound closure, suppressed inflammation, promoted neovascularization and collagen remodeling, and exhibited excellent tissue integration and repair potential. The OBPS system provides a novel and effective treatment for wound healing.
Polydimethylsiloxane (PDMS)-based organ-on-a-chip (OoC) platforms typically rely on cleanroom photolithography, custom masks, and master molds, which limit their accessibility and impede rapid prototyping efforts. Here, we present a simple, mask-free, cleanroom-free method to fabricate membrane-integrated PDMS microfluidic devices using a low-cost digital craft cutter and plasma-assisted adhesive bonding. The process achieves 150µm feature resolution, and forms PDMS-adhesive interfaces with high mechanical robustness (>370 kPa tensile strength) and hydrolytic stability, supporting flow rates up to 20 ml min-1without leakage. Using this method, we fabricated one-lane and two-lane OoC platforms incorporating commercial polycarbonate membranes and lab-made bacterial nanocellulose membranes without requiring membrane modification for bonding. These membrane-integrated devices enabled rapid formation of intestinal tissues within 72 h, significantly faster than conventional PDMS or transwell-based models, which typically require 1-3 weeks to achieve epithelial differentiation. Under physiological shear stress, epithelial tissues exhibited 2-3 fold increase in expression of differentiation markers (Mucin-2, Villin) and substantial reduction in stemness marker expression compared with static culture. In two-lane co-culture systems, epithelial-endothelial interfaces developed functional barrier properties within the same 72 h window and demonstrated membrane-dependent differences in maturation, with nanocellulose membranes promoting enhanced three-dimensional organization. This cleanroom-free platform reduces fabrication barriers for membrane-integrated PDMS OoC devices. Its ability to support rapid, robust intestinal tissue formation makes it a practical platform for accessible organ-mimetic systems in research settings and translational applications.
The diaphragm is a physical barrier separating the thoracic and abdominal cavities. Its physiological function is fundamental to pulmonary ventilation. Congenital diaphragmatic hernia (CDH) is a malformation that leaves a hole in the diaphragm during fetal development. Synthetic nondegradable membranes are currently used for the repair of diaphragm holes. However, these membranes do not help cells to adhere and proliferate. There is a need for biodegradable membranes composed of muscular and fibroblast cells to replicate a simplified diaphragmatic tissue. We thus developed a biocompatible and biodegradable salt-compacted albumin membrane. C2C12 myoblasts were bioprinted as central spokes, surrounded by a ring of NIH 3T3 fibroblasts, onto albumin membranes. We used bioink based on methacrylated collagen and hyaluronic acid, containing porous poly(D,L-lactic-co-glycolic acid) solid microscaffolds to protect myoblast and fibroblast cells against mechanical stress during extrusion printing. We found that metabolic activity of C2C12 myoblast increased by 215% in the presence of a polylysine-coated microscaffolds, compared to those cultured without microscaffolds. Microscaffolds loaded with C2C12 and NIH 3T3 cells increased viability (30% and 15%) and cell activity (527% and 567%) either after co-culture (5.6% increased viability and 588% cell activity) bioprinting on albumin membrane, compared to cells in bioink without microscaffolds. Cell-loaded microscaffolds embedded in bioink enhance C2C12 to NIH3T3 cross-migration on albumin membrane. This work is a preliminary proof of concept of cellularization of myoblasts and fibroblasts by extrusion bioprinting on a new biodegradable albumin membrane designed for diaphragmatic hernia patches.
Skeletal muscle constitutes approximately 40%-50% of total body weight and plays a vital role in human physiology; however, its regenerative capacity is insufficient in cases of volumetric muscle loss, and effective therapeutic options remain limited. To address this challenge, tissue engineering approaches have focused on constructing functional muscle grafts, with increasing attention on vascularized muscle tissues due to their enhanced integration, regeneration, and reduced fibrosis after transplantation. Nevertheless, most existing strategies rely on complex, multi-step fabrication processes requiring external scaffolds or stimulation systems, which limit practical applicability. In this study, we propose a one-step coaxial bioprinting strategy that enables the simultaneous fabrication of aligned muscle and vascular structures within a single construct. By optimizing the bioink formulation, both structural integrity and cellular functionality were achieved. Shear stress generated during coaxial printing induced myoblast alignment, promoting myogenic differentiation, while precise compartmentalization ensured effective domain separation. Furthermore, dynamic culture conditions enhanced endothelial cell maturation and vascular functionality. This scaffold-free and mechanical-stimulus-free vascularized muscle construct provides a simplified yet functional platform with strong potential for muscle disease modeling, drug screening, and regenerative therapies. The proposed fabrication strategy represents a meaningful advancement in vascularized muscle tissue engineering.
Stem cell spheroids, as an advanced platform in cell therapy, have shown great potential for improving therapeutic efficacy through enhanced cell-cell and cell-microenvironment interactions. Meanwhile, zinc-based biomaterials are attracting increasing attention for their biological functions. However, their roles in modulating stem cell spheroids remain largely unclear. In this study, we synthesized zinc-doped urchin-like hydroxyapatite (Zn-uHA) via a hydrothermal method and verified its long-term antibacterial activity. Utilizing microwell array technology, we conducted 3D cell culture of human dental pulp stem cells (hDPSCs) with Zn-uHA, thereby achieving high-throughput and precise fabrication of 3D hDPSCs/Zn-uHA hybrid stem cell spheroids. Compared with pure cell spheroids, hDPSCs/Zn3-uHA hybrid spheroids demonstrated significantly enhanced cell migration and osteogenic differentiation, with osteogenesis-related gene expression upregulated 2.7-fold.In vivostudies further confirmed that the hybrid cell spheroids exhibited favorable biocompatibility, with no evident inflammatory response or immune rejection, and enabled prolonged retention at the implantation site. Additionally, they significantly promoted neovascularization and new bone formation. These findings suggest that Zn-uHA-integrated stem cell spheroids represent a promising strategy for enhancing both antibacterial and osteogenic performance, offering potential applications in bone tissue engineering, particularly in infection-prone clinical scenarios such as orthopedics and dentistry.
The endometrium, a dynamic tissue of paramount importance to female reproductive health, undergoes cyclic remodeling governed by hormonal signals. Endometrial diseases have a relatively high incidence rate and exert a considerable influence on women's quality of life; however, their underlying mechanisms remain incompletely understood. Endometrial organoids (EOs) possess considerable potential to replicate tissue-specific functions and disease phenotypes. This review begins by outlining the physiological and pathological basis of the endometrium, then systematically reviews the latest progress in EOs research, emphasizing the importance of developing precise organoid models. We further discuss strategies for organoid construction, key characteristics, and functional validation methods. Applications of EOs in modeling endometrial diseases, drug screening, and personalized therapy are summarized. Finally, we address current challenges in the field and suggest future directions. Despite challenges related to standardization and clinical translation, continued refinement of EOs systems will deepen our understanding of endometrial biology and accelerate the development of targeted therapies for endometrial diseases.
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 63μm and 96μm at a 300 000 cells ml-1concentration, and 30% for 143µm and 195µm OD nozzles at a 50 000 cells ml-1concentration. Spacing using a 63μm nozzle tests printed droplets with a minimum center-to-center spacing of 199.53 ± 2.52μm on a culture well and cell spacing of 122.23 ± 21.69μm 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.
Urethral strictures and hypospadias remain major challenges in urology, with current treatments limited by donor-site morbidity, graft shrinkage, and poor long-term outcomes. To address these limitations, we developed a biomimetic multilayered hydrogel scaffold with dual cell types and mesh reinforcement for urethral reconstruction. The construct combined predifferentiated buccal mucosa cells and adipose-derived mesenchymal stem cells within a gelatin methacrylate-silk fibroin hydrogel supported by electrowritten and electrospun polymeric meshes, designed to reproduce both epithelial and smooth muscle layers of the native urethra. Implanted into the rabbit urethra and followed for 12 weeks, the scaffold showed favorable integration with early but transient immune activation, preservation of systemic stability, and maintenance of urethral lumen architecture. Histological evaluation revealed the formation of a stratified urothelium and organized smooth muscle bundles with limited fibrosis, highlighting the scaffold's potential to support functional urethral regeneration and serve as an alternative to graft-based urethroplasty.
Rheumatoid arthritis (RA) is a chronic autoimmune inflammatory disorder that afflicts the synovial lining of joints and manifests in reduced range of motion, pain, swelling, and numerous other complications with no effective cure. In recent years, attention has shifted toward eradicating the source of synovial autoimmunity and inflammation, with a key focus on the synovial-draining lymphatics. However, very fewin vitromodels of the synovial microenvironment have been developed to date, and none yet include the synovial lymphatics. We therefore create a microfluidic chip device that models the synovial-draining lymphatics within the subintimal synovium microenvironment. Functional assays on our synovium-on-chip demonstrate increased lymphatic permeability and decreased drainage under RA inflammation compared to healthy controls, accompanied by increased lymphatic endothelial cell (LEC) junctional disruption and altered LEC phenotype following interaction with fibroblast-like synoviocytes. We identify overexpression of chitinase-3 like-protein-1 from RA patient-derived synoviocytes as a key target in inducing junctional loosening and lymphatic dysfunction, which is reversed in our microfluidic chip and in vivo mouse models by neutralizing antibody inhibition. Our novel synovium-on-chip model can provide a physiologically accurate representation of lymphatic drainage and activity under disease conditions, informing tissue engineering, drug testing, and high-throughput screening for RA.
In embedded 3D bioprinting, biomaterial inks are extruded into sacrificial support baths to facilitate the fabrication of complex shapes, even from soft, liquid-like materials. Post-printing, the diffusion of small molecules into or out of the support bath can facilitate ink crosslinking to stabilize the printed structure. In these coupled reaction-diffusion systems, the rheological properties of the ink will change over time. Despite the importance of tuning the mechanical properties of these inks for biological applications, there are currently no methods to accurately predict ink stiffness over time throughout the crosslinking process. Here, we use a custom-developed magnetic stress rheometer to continuously monitor diffusion-driven crosslinking in situ. Our approach reveals how gelation kinetics depend on the thickness of the ink layer, and enables predictive estimation of mechanical evolution in these reaction- and diffusion-driven systems. With these insights, we fabricate specimens with predetermined mechanical properties and observe changes in cell phenotype as a response. These insights help inform the design of inks and timing of bioprinting protocols to achieve prints with desirable mechanical properties and further allow the fabrication of prints with patterned mechanical properties.
Understanding how airborne particulates disrupt the alveolar barrier requiresin vitrosystems that recapitulate both the structure and transport properties of the lung air-blood interface. Here, we report a biodegradable lung alveoli-on-a-chip enabled by porous poly(lactic-co-glycolic acid)/polycaprolactone (PLGA/PCL) membranes with an interconnected porous architecture generated via porogen-assisted phase separation process. The membrane exhibits tunable degradation behavior, allowing progressive increases in surface porosity (∼40%) and reduction in thickness (∼3µm) during culture, while PCL maintains mechanical integrity under dynamic conditions. These degradation-driven structural changes regulate membrane transport properties, leading to enhanced permeability and supporting the formation of a functional epithelial-endothelial barrier under air-liquid interface culture with breathing-mimetic cycling strain. Primary human alveolar epithelial and microvascular endothelial cells formed confluent, junctional monolayers on opposing membrane surfaces, exhibiting stable barrier function and high viability throughout the culture period. As a functional application, the platform was used to assess diesel particulate matter (DPM)-induced alveolar injury. Apical exposure to DPM induced dose-dependent cytotoxicity, increased barrier permeability, elevated reactive oxygen species, and DNA damage in both epithelial and endothelial layers, demonstrating trans-barrier propagation of particulate-induced injury. Pharmacological modulation with roflumilast-N-oxide, a phosphodiesterase-4 inhibitor, selectively attenuated oxidative stress and inflammatory responses, with limited effects on barrier integrity. Together, this work establishes degradable PLGA/PCL membranes as tunable interface materials for lung-on-a-chip systems, where structural evolution during degradation directly governs transport and barrier function. The resulting platform provides a physiologically relevant approach for studying particulate toxicity and therapeutic modulation at the alveolar interface.
Dental implant osseointegration is the process by which these medical devices integrate within bone. Because of the significant failure rate of dental implant osseointegration in the long term, specific models designed to better understand this process are needed. Current limitations related to in vivo models justify the development of in vitro substitutes. The aim of this study was to develop a 3D in vitro mineralized bone model to reproduce dental implant osseointegration. To achieve this, we started by fabricating a 3D mineralized hydrogel-based in vitro model and characterizing it for mineralization, osteogenic differentiation, cell-deposited extracellular matrix (ECM), and inflammatory markers. Immortalized human mesenchymal stem cells were embedded in a methacrylated collagen-hyaluronic acid gel matrix and cultured in osteogenic differentiation medium with elevated calcium concentration. The model exhibited early mineralization, deposition of minerals in a spherical form and expression of osteogenic differentiation markers. Proteomic profiling revealed a collagen-rich ECM with enrichment of type I collagen. Additionally, there was enrichment of signaling pathways involved in osteogenic differentiation. This hydrogel-based model was further integrated into a 3D-printed polylactic acid scaffold with a dental implant as an in vitro platform to study dental implant osseointegration. We demonstrated cellular migration, matrix deposition, and initiation of mineralization on dental implants with two different surface roughnesses. Additionally, a sensitive pull-out test was specifically developed for the model to detect the attachments initiated by the cells. Therefore, the developed model has strong potential to revolutionize dental implant screening by bridging the gap between 2D in vitro models and in vivo models.
Vaginal reconstruction is limited by the lack of biomaterials that replicate the structure, biomechanics, and biochemistry of the native tissues. Synthetic meshes, xenografts, and autologous skin or bowel grafts are hindered by their immunogenicity, poor integration, and non-physiological properties. We report a scalable platform for the fabrication of patient-specific living vaginal grafts from autologous fibroblasts. Using scaffold-free micromolding and automated assembly, fibroblasts from small full-thickness vaginal biopsies self-assembled into a collagen-rich, highly aligned extracellular matrix. To improve mechanical integrity, we create twisted subunit assemblies that displayed highly aligned collagen, dense cellularity, and a predominantly quiescent fibroblast phenotype with minimal myofibroblast activation. This autologous tissue-specific construct addresses the shortcomings of current materials and offers a customizable and biocompatible solution for regenerative gynecology. By combining tissue specificity, immunologic safety, and modular scalability, this approach has the potential to transform surgical options for congenital anomalies, post-oncologic reconstruction, fistula repair, and pelvic organ prolapse.
