Abstract:

Objective To investigate the physicochemical properties of a multifunctional jellyfish collagen （JC） dynamic hydrogel and its effectiveness for three-dimensional （3D） cell culture. Methods The JC-HEC/OHEC-GNP dynamic hydrogels with multiple physical and reversible covalent cross-linking were constructed using JC， hydroxyethyl cellulose （HEC） and its oxidized product （OHEC）， and genipin （GNP）. The physicochemical structures were characterized by ATRFTIR， SEM， XPS， and water contact angle tests. The biological effects on human umbilical vein endothelial cells （HUVECs） and fibroblasts （FBs） were evaluated via CCK-8 assay， Calcein/PI dual staining， EdU assay， tube formation assay， and Transwell migration assay. Through the 3D cell-embedding co-culture experiment， the growth status of HUVECs， FBs and human adipose-derived mesenchymal stem cells （AD-MSCs） in the three-dimensional hydrogel was observed. Results The JC-HEC/OHEC-GNP hydrogels formed a continuous and stable porous 3D network structure. The oxidized JC-OHEC-GNP showed more uniform pores and stronger hydrophilicity. The hydrogels exhibited excellent biocompatibility， significantly promoted the proliferation and migration of HUVECs and FBs， and enhanced the tube formation of HUVECs. Antioxidant assays indicated a significant reduction in reactive oxygen species （ROS） and protection against H2O2-induced oxidative damage. The 3D co-culture showed that HUVECs， FBs， and AD-MSCs grew stably and formed complex dendritic 3D branching networks in the hydrogels， and the number of branching nodes and the volume of the network are significantly better than those of the traditional Matrigel （P<0.05）. Conclusion The JC-HEC/OHEC-GNP dynamic biomimetic hydrogel possesses good physicochemical properties， biocompatibility， antioxidant properties， and pro-angiogenic capabilities， making it a promising biomaterial platform for 3D cell culture and cell therapy in diabetic foot ulcers.

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