Type Ⅰ collagen/sodium alginate/hyaluronic acid composite hydrogel for cell loading and 3D culture in vascular tissue engineering

Abstract

Collagen, sodium alginate and hyaluronic acid are natural-derived polymer materials with good cell compatibility and bio-safety, which are widely used in cell culture, tissue engineering and drug delivery, and so on. Pure collagen has poor mechanical properties. When preparing collagen and sodium alginate to form a composite hydrogel material, the mechanical properties and porosity of the hydrogel scaffold can be improved by adjusting the degree of cross-linking of sodium alginate and Ca2+ mimicking extracellular matrix. The Young's modulus and the sol-gel transition temperature of the hydrogel were characterized by PIUMA nanoindenter and DHR rheometer in this study. Microscopic images of endothelial cells expressing red fluorescent proteins and mesenchymal stem cell expressing green fluorescent proteins were captured with Olympus fluorescence microscope after cell cultured for 0 day, 3 days, 5 days and 7 days in hybrid hydrogel microenvironment, and the images of endothelial cell spheroid growth diffusion after cell cultured for 1 day, 6 days and 9 days. The results show that the hybrid hydrogel is cytocompatible. The Young's modulus of the hydrogel is (600±81) Pa and its sol-gel transition temperature is 23.2℃. In conclusion, type Ⅰ collagen/sodium alginate/hyaluronic acid hydrogel has good cytocompatibility for endothelial cells and mesenchymal stem cells, and can be used as an ideal scaffold material for cell 3D culture. The Young's modulus and sol-gel transition temperature of the hydrogel have no damage to cell viability, which can be used as an in vitro model for studying angiogenesis and has important application prospects in vascular tissue engin-eering.

Keywords

• 3d cell culture
• type ⅰ collagen
• sodium alginate
• hydrogel microenvironment