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A novel cell encapsulatable cryogel (CECG) with macro-porous structures and high permeability: a three-dimensional cell culture scaffold for enhanced cell adhesion and proliferation

Hydrogel scaffold is a popular cell delivery vehicle in tissue engineering and regenerative medicine due to its capability to encapsulate cells as well as its modifiable properties. However, the inherent submicron- or nano-sized polymer networks of conventional hydrogel will produce spatial constraints on cellular activities of encapsulated cells. In this study, we endeavor to develop an innovative cell encapsulatable cryogel (CECG) platform with interconnected macro-pores, by combining cell cryopreservation technique with cryogel preparation process. The hyaluronan (HA) CECG constructs are fabricated under the freezing conditions via UV photo-crosslinking of the HA methacrylate (HA-MA) that are dissolved in the ‘freezing solvent’, namely the phosphate buffered saline supplemented with dimethyl sulphoxide and fetal bovine serum. Two model cell types, chondrocytes and human mesenchymal stem cells (hMSCs), can be uniformly three-dimensionally encapsulated into HA CECG constructs with high cell viability, respectively. The macro-porous structures, generated from phase separation under freezing, endow HA CECG constructs with higher permeability and more living space for cell growth. The chondrocytes encapsulated in HA CECG possess enhanced proliferation and extracellular matrix secretion than those in conventional HA hydrogels. In addition, the HA-Gel CECG constructs, fabricated with HA-MA and gelatin methacrylate precursors, provide cell-adhesive interfaces to facilitate hMSCs attachment and proliferation. The results of this work may lay the foundation for us to explore the applications of the CECG-based scaffolds in the field of tissue engineering and regenerative medicine.