Electrostatic Droplet Generator with 3-Coaxial-Nozzle Head for Microencapsulation of Living Cells in Hydrogel Covered by Synthetic Polymer Membranes

The current methods allow for encapsulation of cells inside spherical microcapsules made of a matrix covered by a permselective membrane using an electrostatic droplet generator with 1-nozzle or 2-nozzle heads. However, some potentially useful materials for the outer membranes cannot be put into direct contact with hydrophilic core filled by cells during the manufacturing process. Therefore, we designed a novel 3-coaxial-nozzle head that allows for the third fluid to separate the core material from the membrane material. The equipment was applied for manufacturing spherical microcapsules comprised of cell-friendly alginate core surrounded by semipermeable polyethersulfone membrane. The obtained microcapsules had a diameter between 0.84 mm and 1.79 mm, and the diameter correlated negatively with the applied electric voltage. The thickness of the membrane varied from 171 µm to 450 µm. The SEM images of the interior of microcapsules revealed highly porous membrane structure typical for synthetic membranes obtained by a wet phase inversion method. Bakery yeast cells encapsulated inside the alginate-polyethersulfone microcapsules retained their proliferation ability proving the effectiveness and safety of this encapsulation technique.     

Chitosan Hydrogel Supplemented with Metformin Promotes Neuron–like Cell Differentiation of Gingival Mesenchymal Stem Cells

Human gingival mesenchymal stem cells (GMSCs) are derived from migratory neural crest stem cells and have the potential to differentiate into neurons. Metformin can inhibit stem–cell aging and promotes the regeneration and development of neurons. In this study, we investigated the potential of metformin as an enhancer on neuronal differentiation of GMSCs in the growth environment of chitosan hydrogel. The crosslinked chitosan/β–glycerophosphate hydrogel can form a perforated microporous structure that is suitable for cell growth and channels to transport water and macromolecules. GMSCs have powerful osteogenic, adipogenic and chondrogenic abilities in the induction medium supplemented with metformin. After induction in an induction medium supplemented with metformin, Western blot and immunofluorescence results showed that GMSCs differentiated into neuron–like cells with a significantly enhanced expression of neuro–related markers, including Nestin (NES) and β–Tubulin (TUJ1). Proteomics was used to construct protein profiles in neural differentiation, and the results showed that chitosan hydrogels containing metformin promoted the upregulation of neural regeneration–related proteins, including ATP5F1, ATP5J, NADH dehydrogenase (ubiquinone) Fe–S protein 3 (NDUFS3), and Glutamate Dehydrogenase 1 (GLUD1). Our results help to promote the clinical application of stem–cell neural regeneration.     

Bioactive and Biocompatible Macroporous Scaffolds with Tunable Performances Prepared Based on 3D Printing of the Pre‐Crosslinked Sodium Alginate/Hydroxyapatite Hydrogel Ink

Bioactive and biocompatible porous scaffold materials with adjustable pore structures and drug delivery capability are one of the key elements in bone tissue engineering. In this work, bioactive and biocompatible sodium alginate (SA)/hydroxyapatite (HAP) macroporous scaffolds are facilely and effectively fabricated based on 3D printing of the pre‐crosslinked SA/HAP hydrogels followed by further crosslinking to improve the mechanical properties of scaffolds. The pore structures and porosity (>80%) of the porous scaffolds can be readily tailored by varying the formation conditions. Furthermore, the in vitro biomineralization tests show that the bioactivity of the porous scaffolds is effectively enhanced by the addition of HAP nanoparticles into the scaffold matrix. Furthermore, the anti‐inflammatory drug curcumin is loaded into the porous scaffolds and the in vitro release study shows the sustainable drug release function of the porous scaffolds. Moreover, mouse bone mesenchymal stem cells (mBMSCs) are cultured on the porous scaffolds, and the results of the in vitro biocompatibility experiment show that the mBMSCs can be adhered well on the porous scaffolds. All of the results suggest that the bioactive and biocompatible SA/HAP porous scaffolds have great application potential in bone tissue engineering.