Engineering Soft Spring Gauges for In Situ Biomaterial and Tissue Weighing.

Hydrogels have gained great attention and broad applications in tissue engineering, regenerative medicine, and drug delivery due to their excellent biocompatibility and degradability. However, accurately and noninvasively characterizing the degradation process of hydrogels remains a challenge. To address this, we have developed a method using soft spring gauges (SSGs) for the in situ weighing of hydrogels. Our approach uses a simple hydrogel-based sacrificial template method to fabricate polydimethylsiloxane (PDMS) SSGs. The SSGs used in this study can characterize hydrogels with a minimum wet weight of approximately 30 mg. Through theoretical derivations, numerical simulations, and experimental characterization, we confirmed that the length change of the SSGs in a buffer solution correlates linearly with the applied hanging weights. This allows us to track and assess the solid mass change of hydrogels during degradation with high feasibility and accuracy. Additionally, we have demonstrated the potential application of SSGs for the in situ characterization of engineered tissue growth. This method represents an advanced approach for in situ hydrogel weighing, holding great promise for advancing the development of hydrogels and other biomaterials in biomedical applications.

Bioinspired Microstructure Platform for Modular Cell-Laden Microgel Fabrication.

Cell-laden microgels have attracted increasing interest in various biomedical fields, as living building blocks to construct spatially organized multicellular structures or complex tissue features (e.g., cell spheroids and aligned cells/fibers). Although numerous approaches have been developed to tailor cell-laden microgels, there is still an unmet need for modular, versatile, convenient, and high-throughput methods. In this study, as inspired by the phenomena of water droplet manipulation from natural microstructures, a novel platform is developed to manipulate microscale hydrogel droplets and fabricate modular cell-laden microgels. First, taking antenna-like trichome as a template, catcher-like bioinspired microstructures are fabricated and hydrogel droplets are manipulated modularly in a versatile, convenient, and high-throughput manner, which is compatible with various types of hydrogels (e.g., photo-cross-linking, thermal-cross-linking, and ion-cross-linking). It is demonstrated that this platform can manipulate cell-laden microgels as modular units, such as two or more cell-laden microgels on one single catcher-like structure and different structures on one single chip. The authors also demonstrate the application of this platform on constructing complex tissue features like myocardial fibrosis tissue models to study cardiac fibrosis. The developed platform will be a powerful tool for engineering various in vitro tissue models for widespread biomedical applications.

Fluorescent conjugated polymer nanovector for in vivo tracking and regulating the fate of stem cells for restoring infarcted myocardium.

Stem cell therapy holds great promise for cardiac regeneration. However, the lack of ability to control stem cell fate after in vivo transplantation greatly restricts its therapeutic outcomes. MicroRNA delivery has emerged as a powerful tool to control stem cell fate for enhanced cardiac regeneration. However, the clinical translation of therapy based on gene-transfected stem cells remains challenging, due to the unknown in vivo behaviors of stem cells. Here, we developed a nano-platform (i.e., PFBT@miR-1-Tat NPs) that can achieve triggered release of microRNA-1 to promote cardiac differentiation of mesenchymal stem cells (MSCs), and long-term tracking of transplanted MSCs through bright and ultra-stable fluorescence of conjugated polymer poly(9,9-dioctylfluorene-alt-benzothiadiazole) (PFBT). We found that PFBT@miR-1-Tat NP-treated MSCs significantly restored the infarcted myocardium by promoting stem cell cardiac differentiation and integration with the in situ cardiac tissues. Meanwhile, MSCs without gene delivery improved the infarcted heart functions mainly through a paracrine effect and blood vessel formation. The developed conjugated polymer nanovector should be a powerful tool for manipulating as well as revealing the fate of therapeutic cells in vivo, which is critical for optimizing the therapeutic route of gene and cell combined therapy and therefore for accelerating clinical translation. STATEMENT OF SIGNIFICANCE: The lack of controllability in stem cell fate and the unclear in vivo cellular behaviors restrict the therapeutic outcomes of stem cell therapy. Herein, we engineered fluorescent conjugated polymer nanoparticles as gene delivery nanovectors with controlled release and high intracellular delivery capability to harness the fate of mesenchymal stem cells (MSCs) in vivo, meanwhile to reveal the cellular mechanism of gene-treated stem cell therapy. As compared with only MSC treatment that improves infarcted myocardium functions through paracrine effect, treatment with conjugated polymer nanovector-treated MSCs significantly restored infarcted myocardium through enhancing MSC cardiac differentiation and integration with the in-situ cardiac tissues. These findings demonstrate that the conjugated polymer nanovector would be a powerful tool in optimizing gene and cell combined therapy.

Engineering ellipsoidal cap-like hydrogel particles as building blocks or sacrificial templates for three-dimensional cell culture.

Hydrogel particles that can be engineered to compartmentally culture cells in a three-dimensional (3D) and high-throughput manner have attracted increasing interest in the biomedical area. However, the ability to generate hydrogel particles with specially designed structures and their potential biomedical applications need to be further explored. This work introduces a method for fabricating hydrogel particles in an ellipsoidal cap-like shape (i.e., ellipsoidal cap-like hydrogel particles) by employing an open-pore anodic aluminum oxide membrane. Hydrogel particles of different sizes are fabricated. The ability to produce ellipsoidal cap-like magnetic hydrogel particles with controlled distribution of magnetic nanoparticles is demonstrated. Encapsulated cells show high viability, indicating the potential for using these hydrogel particles as structure- and remote-controllable building blocks for tissue engineering application. Moreover, the hydrogel particles are also used as sacrificial templates for fabricating ellipsoidal cap-like concave wells, which are further applied for producing size controllable cell aggregates. The results are beneficial for the development of hydrogel particles and their applications in 3D cell culture.

Bioprinting 3D cell-laden hydrogel microarray for screening human periodontal ligament stem cell response to extracellular matrix.

Periodontitis is an inflammatory disease negatively affecting up to 15% of adults worldwide. Periodontal ligament stem cells (PDLSCs) hold great promises for periodontal tissue regeneration, where it is necessary to find proper extracellular matrix (ECM) materials (e.g., composition, concentration). In this study, we proposed a bioprinting-based approach to generate nano-liter sized three-dimensional (3D) cell-laden hydrogel array with gradient of ECM components, through controlling the volume ratio of two hydrogels, such as gelatin methacrylate (GelMA) and poly(ethylene glycol) (PEG) dimethacrylate. The resulting cell-laden array with a gradient of GelMA/PEG composition was used to screen human PDLSC response to ECM. The behavior (e.g., cell viability, spreading) of human PDLSCs in GelMA/PEG array were found to be depended on the volume ratios of GelMA/PEG, with cell viability and spreading area decreased along with increasing the ratio of PEG. The developed approach would be useful for screening cell-biomaterial interaction in 3D and promoting regeneration of functional tissue.

Fabrication of Microscale Hydrogels with Tailored Microstructures based on Liquid Bridge Phenomenon.

Microscale hydrogels (microgels) find widespread applications in various fields, such as drug delivery, tissue engineering, and biosensing. The shape of the microgels is a critical parameter that can significantly influence their function in these applications. Although various methods have been developed (e.g., micromolding, photolithography, microfluidics, and mechanical deformation method), it is still technically challenging to fabricate microgels with tailored microstructures. In this study, we have developed a simple and versatile method for preparing microgels by stretching hydrogel precursor droplets between two substrates to form a liquid bridge. Microgels with tailored microstructures (e.g., barrel-like, dumbbell-like, or funnel-like shapes) have been achieved through adjusting the distance between and the hydrophobicity of the two substrates. The developed method holds great potential to impact multiple fields, such as drug delivery, tissue engineering, and biosensing.

Benchtop fabrication of three-dimensional reconfigurable microfluidic devices from paper-polymer composite.

We presented a benchtop technique that can fabricate reconfigurable, three-dimensional (3D) microfluidic devices made from a soft paper-polymer composite. This fabrication approach can produce microchannels at a minimal width of 100 µm and can be used to prototype 3D microfluidic devices by simple bending and stretching. The entire fabrication process can be finished in 2 hours on a laboratory bench without the need for special equipment involved in lithography. Various functional microfluidic devices (e.g., droplet generator and reconfigurable electronic circuit) were prepared using this paper-polymer hybrid microfluidic system. The developed method can be applied in a wide range of standard applications and emerging technologies such as liquid-phase electronics.