Alginate Hydrogel Microtubes for Salivary Gland Cell Organization and Cavitation

Understanding the different regulatory functions of epithelial and mesenchymal cell types in salivary gland development and cellular organization is essential for proper organoid formation and salivary gland tissue regeneration. Here, we demonstrate a biocompatible platform using pre-formed alginate hydrogel microtubes to facilitate direct epithelial–mesenchymal cell interaction for 3D salivary gland cell organization, which allows for monitoring cellular organization while providing a protective barrier from cell-cluster loss during medium changes. Using mouse salivary gland ductal epithelial SIMS cells as the epithelial model cell type and NIH 3T3 fibroblasts or primary E16 salivary mesenchyme cells as the stromal model cell types, self-organization from epithelial–mesenchymal interaction was examined. We observed that epithelial and mesenchymal cells undergo aggregation on day 1, cavitation by day 4, and generation of an EpCAM-expressing epithelial cell layer as early as day 7 of the co-culture in hydrogel microtubes, demonstrating the utility of hydrogel microtubes to facilitate heterotypic cell–cell interactions to form cavitated organoids. Thus, pre-formed alginate microtubes are a promising co-culture method for further understanding epithelial and mesenchymal interaction during tissue morphogenesis and for future practical applications in regenerative medicine.

Droplet-Based Microfluidic Preparation of Shape-Variable Alginate Hydrogel Magnetic Micromotors

This article introduces a facile droplet-based microfluidic method for the preparation of Fe3O4-incorporated alginate hydrogel magnetic micromotors with variable shapes. By using droplet-based microfluidics and water diffusion, monodisperse (quasi-)spherical microparticles of sodium alginate and Fe3O4 (Na-Alg/Fe3O4) are obtained. The diameter varies from 31.9 to 102.7 µm with the initial concentration of Na-Alginate in dispersed fluid ranging from 0.09 to 9 mg/mL. Calcium chloride (CaCl2) is used for gelation, immediately transforming Na-Alg/Fe3O4 microparticles into Ca-Alginate hydrogel microparticles incorporating Fe3O4 nanoparticles, i.e., Ca-Alg/Fe3O4 micromotors. Spherical, droplet-like, and worm-like shapes are yielded depending on the concentration of CaCl2, which is explained by crosslinking and anisotropic swelling during the gelation. The locomotion of Ca-Alg/Fe3O4 micromotors is activated by applying external magnetic fields. Under the rotating magnetic field (5 mT, 1–15 Hz), spherical Ca-Alg/Fe3O4 micromotors exhibit an average advancing velocity up to 158.2 ± 8.6 µm/s, whereas worm-like Ca-Alg/Fe3O4 micromotors could be rotated for potential advancing. Under the magnetic field gradient (3 T/m), droplet-like Ca-Alg/Fe3O4 micromotors are pulled forward with the average velocity of 70.7 ± 2.8 µm/s. This article provides an inspiring and timesaving approach for the preparation of shape-variable hydrogel micromotors without using complex patterns or sophisticated facilities, which holds potential for biomedical applications such as targeted drug delivery.

Encapsulated Allogeneic Synovial Membrane Mesenchymal Stem Cells Provide Better Outcomes of Chondral Lesions in Horses

Osteoarthritis is the main cause of equine lameness, and its treatment remains ineffective. Synovial membrane mesenchymal stem cells (SMMSCs) provide satisfactory outcomes in joint injuries, mainly due to their immunomodulatory and reparative properties. This study aimed to evaluate the effect of SMMSCs, either encapsulated in alginate hydrogel or free, in chondral lesions of horses.Methods: Chondral lesions were surgically induced in the medial trochlea of the talus of fifteen horses. Animals were treated with PBS free SMMSCs or encapsulated SMMSCs. Physical evaluations, assignment of lameness scores and synovial fluid analysis were performed (cytological analysis and dosage of IL-1, IL-10, IL-6, INF-Ɣ, TNF 𝛼, P substance, serum amyloid A, TGF-β, IGF and PGE2) for two weeks. Cartilage biopsies were performed 150 days after induction for histological analysis and immunohistochemistry staining.Results: All groups initially presented inflammation. Although free SMMSCs showed moderate tissue repair, encapsulated SMMSCs had a lower grade of inflammation with superior tissue macro- and microscopic aspects at the end, while the control group showed fibrosis and poor cartilage aspect. This study suggests better effectiveness of stem cells in chondral defects when encapsulated MSCs are used.Conclusion: While the absence of treatment perpetuates cartilage degradation, encapsulated SMMSCs respond better to initial inflammation, interacting and modulating the environment through the release of anti-inflammatory cytokines. Better outcomes observed in encapsulated MSCs were related to the immuno- and physical barriers provided by the alginate hydrogel, allowing a longer period of permanence and interaction between MSCs and the environment.

Advanced Strategies for 3D Bioprinting of Tissue and Organs Analogs Using Alginate Hydrogel Bioinks

Alginate is a natural polysaccharide that typically originates from various species of algae. Due to its low cost, good biocompatibility, and rapid ionic gelation, the alginate hydrogel has become a good option of bioink source for 3D bioprinting. However, the lack of cell adhesive moieties, erratic biodegradability, and poor printability are the critical limitations of alginate hydrogel bioink. This review discusses the pivotal properties of alginate hydrogel as a bioink for 3D bioprinting technologies. Afterward, a variety of advanced material formulations and biofabrication strategies that have recently been developed to overcome the drawbacks of alginate hydrogel bioink will be focused on. In addition, the applications of these advanced solutions for 3D bioprinting of tissue/organ mimicries such as regenerative implants and in vitro tissue models using alginate-based bioink will be systematically summarized.

3D-Printed Coaxial Hydrogel Patches with Mussel-Inspired Elements for Prolonged Release of Gemcitabine

With the aim of fabricating drug-loaded implantable patches, a 3D printing technique was employed to produce novel coaxial hydrogel patches. The core-section of these patches contained a dopamine-modified methacrylated alginate hydrogel loaded with a chemotherapeutic drug (Gemcitabine), while their shell section was solely comprised of a methacrylated alginate hydrogel. Subsequently, these patches were further modified with CaCO3 cross linker and a polylactic acid (PLA) coating to facilitate prolonged release of the drug. Consequently, the results showed that addition of CaCO3 to the formula enhanced the mechanical properties of the patches and significantly reduced their swelling ratio as compared to that for patches without CaCO3. Furthermore, addition of PLA coating to CaCO3-containing patches has further reduced their swelling ratio, which then significantly slowed down the release of Gemcitabine, to a point where 4-layered patches could release the drug over a period of 7 days in vitro. Remarkably, it was shown that 3-layered and 4-layered Gemcitabine loaded patches were successful in inhibiting pancreatic cancer cell growth for a period of 14 days when tested in vitro. Lastly, in vivo experiments showed that gemcitabine-loaded 4-layered patches were capable of reducing the tumor growth rate and caused no severe toxicity when tested in mice. Altogether, 3D printed hydrogel patches might be used as biocompatible implants for local delivery of drugs to diseased site, to either shrink the tumor or to prevent the tumor recurrence after resection.