Precise liquid handling is an essential operation for self-driving laboratories. In 2023, we introduced the digital pipette, a low-cost, 3D-printed device that enables accurate liquid transfer by robotic arms. However, the initial version lacked mechanisms to prevent cross-contamination when handling multiple liquids. In this commit paper, we present the digital pipette v2, an updated design that mitigates contamination risk by allowing robotic arms to exchange pipette tips. The new hardware achieves liquid handling accuracy within the permissible error range defined by ISO 8655-2, supporting a broader range of experiments involving multiple liquids.

Automating biological experimentation remains challenging due to the need for millimeter-scale precision and long, multi-step workflows and the dynamic nature of living systems. Current liquid handlers only partially automate workflows, requiring human intervention for plate loading, tip replacement, and calibration, while industrial systems are costly and lack flexibility for research settings. Meanwhile, research in autonomous robotics has yet to bridge the gap for long-duration, failure-sensitive biological experiments. Here, we report RoboCulture, a cost-effective and flexible platform that uses a general-purpose robotic manipulator to automate key biological tasks. RoboCulture performs liquid handling, interacts with laboratory equipment, and leverages computer vision for real-time decision-making based on optical density monitoring. We demonstrate a fully autonomous 15-h yeast culture experiment in which RoboCulture uses vision and force feedback together with a modular behavior tree framework to robustly execute, monitor, and manage experiments.

The epicardium is an essential regulator of cardiac development, homeostasis, and injury, yet the composition and impacts of epicardial cell-secreted extracellular vesicles (EVs) remain incompletely understood. Here, we harness an epicardial Biowire platform integrating human stem cell derived epicardial cells with functional myocardium and apply transcriptomics to reveal enriched EV transport in tissues containing epicardial cells. Profiling epicardial-EVs identified key miRNAs and their conservation through stimulated epithelial-to-mesenchymal transition. Supplementation of epicardial-EVs to tissues undergoing ischemia-reperfusion injury influenced gene expression associated with reduction of extracellular matrix remodeling, fibroblast activation, and suppression of cell-ECM interactions. Correlation of EV-miRNAs with mRNA targets highlighted the role of miR-30d-5p, miR-9-5p, miR-16-5p, and the let-7 family in moderating deleterious fibrotic activation and matrix remodeling during myocardial injury in vitro.

In their native environment, tissues are organized into intricate fractal structures, rarely recapitulated in their culture in vitro. The extent to which fractal (self-similar) patterns that resemble complex topography in vivo influence cell maturation remain inadequately elucidated. Yet, the application of fractal topographical stimulation may address the challenge of improving the differentiated cell phenotype in vitro. Here, we show fractality in the kidney glomerulus and podocytes, branching highly differentiated cells within the glomerulus. Biomimetic fractal patterns derived from glomerular histology are used to generate topographical (2.5-D) substrates for cell culture. Podocytes grown on fractal topography exhibit higher expression of functional markers and enhanced cell polarity. RNA sequencing suggests podocytes' enhanced ECM deposition and remodeling on fractal versus flat topography, and enhanced maturation accompanied by stress on fractal versus non-fractal topography. The incorporation of fractal topography into standard well plates may serve as a user-friendly bioengineered platform for high-fidelity cell culture.

Automating biological experimentation remains challenging due to the need for millimeter-scale precision, long and multi-step experiments, and the dynamic nature of living systems. Current liquid handlers only partially automate workflows, requiring human intervention for plate loading, tip replacement, and calibration. Industrial solutions offer more automation but are costly and lack the flexibility needed in research settings. Meanwhile, research in autonomous robotics has yet to bridge the gap for long-duration, failure-sensitive biological experiments. We introduce RoboCulture, a cost-effective and flexible platform that uses a general-purpose robotic manipulator to automate key biological tasks. RoboCulture performs liquid handling, interacts with lab equipment, and leverages computer vision for real-time decisions using optical density-based growth monitoring. We demonstrate a fully autonomous 15-hour yeast culture experiment where RoboCulture uses vision and force feedback and a modular behavior tree framework to robustly execute, monitor, and manage experiments.

Application of cardiac patches to the heart surface can be undertaken to provide support and facilitate regeneration of the damaged cardiac tissue following ischemic injury. Biomaterial composition is an important consideration in the design of cardiac patch materials as it governs host response to ultimately prevent the undesirable fibrotic response. Here, we investigate a novel patch material, poly (itaconate-co-citrate-co-octanediol) (PICO), in the context of cardiac implantation. Citric acid (CA) and itaconic acid (ITA), the molecular components of PICO, provided a level of protection for cardiac cells during ischemic reperfusion injury in vitro. Biofabricated PICO patches were shown to degrade in accelerated and hydrolytic conditions, with CA and ITA being released upon degradation. Furthermore, the host response to PICO patches after implantation on rat epicardium in vivo was explored and compared to two biocompatible cardiac patch materials, poly (octamethylene (anhydride) citrate) (POMaC) and poly (ethylene glycol) diacrylate (PEGDA). PICO patches resulted in less macrophage infiltration and lower foreign body giant cell reaction compared to the other materials, with corresponding reduction in smooth muscle actin-positive vessel infiltration into the implant region. Overall, this work demonstrates that PICO patches release CA and ITA upon degradation, both of which demonstrate cardioprotective effects on cardiac cells after ischemic injury, and that PICO patches generate a reduced inflammatory response upon implantation to the heart compared to other materials, signifying promise for use in cardiac patch applications.

Epicardial cells (EPIs) form the outer layer of the heart and play an important role in development and disease. Current heart-on-a-chip platforms still do not fully mimic the native cardiac environment due to the absence of relevant cell types, such as EPIs. Here, using the Biowire II platform, engineered cardiac tissues with an epicardial outer layer and inner myocardial structure are constructed, and an image analysis approach is developed to track the EPI cell migration in a beating myocardial environment. Functional properties of EPI cardiac tissues improve over two weeks in culture. In conditions mimicking ischemia reperfusion injury (IRI), the EPI cardiac tissues experience less cell death and a lower impact on functional properties. EPI cell coverage is significantly reduced and more diffuse under normoxic conditions compared to the post-IRI conditions. Upon IRI, migration of EPI cells into the cardiac tissue interior is observed, with contributions to alpha smooth muscle actin positive cell population. Altogether, a novel heart-on-a-chip model is designed to incorporate EPIs through a formation process that mimics cardiac development, and this work demonstrates that EPI cardiac tissues respond to injury differently than epicardium-free controls, highlighting the importance of including EPIs in heart-on-a-chip constructs that aim to accurately mimic the cardiac environment.

Despite current efforts in organ‐on‐chip engineering to construct miniature cardiac models, they often lack some physiological aspects of the heart, including fiber orientation. This motivates the development of bioartificial left ventricle models that mimic the myofiber orientation of the native ventricle. Herein, an approach relying on microfabricated elastomers that enables hierarchical assembly of 2D aligned cell sheets into a functional conical cardiac ventricle is described. Soft lithography and injection molding techniques are used to fabricate micro‐grooves on an elastomeric polymer scaffold with three different orientations ranging from −60° to +60°, each on a separate trapezoidal construct. The width of the micro‐grooves is optimized to direct the majority of cells along the groove direction and while periodic breaks are used to promote cell–cell contact. The scaffold is wrapped around a central mandrel to obtain a conical‐shaped left ventricle model inspired by the size of a human left ventricle 19 weeks post‐gestation. Rectangular micro‐scale holes are incorporated to alleviate oxygen diffusional limitations within the 3D scaffold. Cardiomyocytes within the 3D left ventricle constructs showed high viability in all layers after 7 days of cultivation. The hierarchically assembled left ventricle also provided functional readouts such as calcium transients and ejection fraction.

Bioelastomers are extensively used in biomedical applications due to their desirable mechanical strength, tunable properties, and chemical versatility; however, three-dimensional (3D) printing bioelastomers into microscale structures has proven elusive. Herein, a high throughput omnidirectional printing approach via coaxial extrusion is described that fabricates perfusable elastomeric microtubes of unprecedently small inner diameter (350-550 µm) and wall thickness (40-60 µm). The versatility of this approach is shown through the printing of two different polymeric elastomers, followed by photocrosslinking and removal of the fugitive inner phase. Designed experiments are used to tune the microtube dimensions and stiffness to match that of native ex vivo rat vasculature. This approach affords the fabrication of multiple biomimetic shapes resembling cochlea and kidney glomerulus and affords facile, high-throughput generation of perfusable structures that can be seeded with endothelial cells for biomedical applications. Post-printing laser micromachining is performed to generate micro-sized holes (520 µm) in the tube wall to tune microstructure permeability. Importantly, for organ-on-a-chip applications, the described approach takes only 3.6 min to print microtubes (without microholes) over an entire 96-well plate device, in contrast to comparable hole-free structures that take between 1.5 and 6.5 days to fabricate using a manual 3D stamping approach.

Epithelial-to-mesenchymal transition (EMT) is a process that occurs in a wide range of tissues and environments, in response to numerous factors and conditions, and plays a critical role in development, disease, and regeneration. The process involves epithelia transitioning into a mobile state and becoming mesenchymal cells. The investigation of EMT processes has been important for understanding developmental biology and disease progression, enabling the advancement of treatment approaches for a variety of disorders such as cancer and myocardial infarction. More recently, tissue engineering efforts have also recognized the importance of controlling the EMT process. In this review, we provide an overview of the EMT process and the signaling pathways and factors that control it, followed by a discussion of bioengineering strategies to control EMT. Important biological, biomaterial, biochemical, and physical factors and properties that have been utilized to control EMT are described, as well as the studies that have investigated the modulation of EMT in tissue engineering and regenerative approaches in vivo, with a specific focus on the heart. Novel tools that can be used to characterize and assess EMT are discussed and finally, we close with a perspective on new bioengineering methods that have the potential to transform our ability to control EMT, ultimately leading to new therapies.

Bioelastomers have been extensively used in tissue engineering applications because of favorable mechanical stability, tunable properties, and chemical versatility. As these materials generally possess low elastic modulus and relatively long gelation time, it is challenging to 3D print them using traditional techniques. Instead, the field of 3D printing has focused preferentially on hydrogels and rigid polyester materials. To develop a versatile approach for 3D printing of elastomers, we used freeform reversible embedding of suspended prepolymers. A family of novel fast photocrosslinakble bioelastomer prepolymers were synthesized from dimethyl itaconate, 1,8-octanediol, and triethyl citrate. Tensile testing confirmed their elastic properties with Young's moduli in the range of 11-53 kPa. These materials supported cultivation of viable cells and enabled adhesion and proliferation of human umbilical vein endothelial cells. Tubular structures were created by embedding the 3D printed microtubes within a secondary hydrogel that served as a temporary support. Upon photocrosslinking and porogen leaching, the polymers were permeable to small molecules (TRITC-dextran). The polymer microtubes were assembled on the 96-well plates custom made by hot-embossing, as a tool to connect multiple organs-on-a-chip. The endothelialization of the tubes was performed to confirm that these microtubes can be utilized as vascular tubes to support parenchymal tissues seeded on them.

In order to secure biomaterials to tissue surfaces, sutures or glues are commonly used. Of interest is the development of a biomaterial patch for applications in tissue engineering and regeneration that incorporates an adhesive component to simplify patch application and ensure sufficient adhesion. A separate region dedicated to fulfilling the specific requirements of an application such as mechanical support or tissue delivery is also desirable. Here, the design and fabrication of a unique patch are presented with distinct regions for adhesion and function, resulting in a biomaterial patch resembling the Band-Aid. The adhesive region contains a novel polymer, synthesized to incorporate a molecule capable of adhesion to tissue, dopamine. The desired polymer composition for patch development is selected based on chemical assessment and evaluation of key physical properties such as swelling and elastic modulus, which are tailored for use in soft tissue applications. The selected polymer formulation, referred to as the adhesive patch (AP) polymer, demonstrates negligible cytotoxicity and improves adhesive capability to rat cardiac tissue compared to currently used patch materials. Finally, the AP polymer is used in the patch, designed to possess distinct adhesive and nonadhesive domains, presenting a novel design for the next generation of biomaterials.

In article number 2000046, Milica Radisic and co-workers design a biomaterial patch that incorporates an adhesive component to improve attachment. To achieve this, a novel polymer is synthesized with elastic properties tailored for cardiac patch applications and enhanced adhesion to tissue. In terms of application, this material is used to fabricate a dual-component patch with distinct regions for adhesion and function.

Itaconate (ITA) is an emerging powerhouse of innate immunity with therapeutic potential that is limited in its ability to be administered in a soluble form. We developed a library of polyester materials that incorporate ITA into polymer backbones resulting in materials with inherent immunoregulatory behavior. Harnessing hydrolytic degradation release from polyester backbones, ITA polymers resulted in the mechanism specific immunoregulatory properties on macrophage polarization in vitro. In a functional assay, the polymer-released ITA inhibited bacterial growth on acetate. Translation to an in vivo model of biomaterial associated inflammation, intraperitoneal injection of ITA polymers demonstrated a rapid resolution of inflammation in comparison to a control polymer silicone, demonstrating the value of sustained biomimetic presentation of ITA.

Synthetic polyester elastomeric constructs have become increasingly important for a range of healthcare applications, due to tunable soft elastic properties that mimic those of human tissues. A number of these constructs require intricate mechanical design to achieve a tunable material with controllable curing. Here, the synthesis and characterization of poly(itaconate-co-citrate-co-octanediol) (PICO) is presented, which exhibits tunable formation of elastomeric networks through radical crosslinking of itaconate in the polymer backbone of viscous polyester gels. Through variation of reaction times and monomer molar composition, materials with modulation of a wide range of elasticity (36-1476 kPa) are generated, indicating the tunability of materials to specific elastomeric constructs. This correlated with measured rapid and controllable gelation times. As a proof of principle, scaffold support for cardiac tissue patches is developed, which presents visible tissue organization and viability with appropriate elastomeric support from PICO materials. These formulations present potential application in a range of healthcare applications with requirement for elastomeric support with controllable, rapid gelation under mild conditions.

viscoelastic adhesive cardiac patch with optimal mechanical behaviour, determined using a computational model, restores heart function and slows down pathological remodelling following myocardial infarction in rodents.