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2022, Vol. 26 ›› Issue (16): 2532-2539Cell-carrying porous methacrylate anhydride gelatin three-dimensional scaffolds and their effects on cell behaviorLi Mingxin1, Li Jun2, Wang Wenchao2, Song Ping3, Lei Haoyuan3, Gui Xingyu3, Zhang Chengyun2, Zhou Changchun3, Liu Lei2
Abstract: BACKGROUND: Hydrogel scaffolds are one of the ideal materials for three-dimensional cell culture. However, the dense network structure in the hydrogel can significantly inhibit the proliferation and extension of cells, and the porous structure in the hydrogel scaffolds can alleviate these problems. OBJECTIVE: To investigate the method of constructing three-dimensional cellular porous hydrogel scaffold with methacrylate anhydride gelatin as scaffold material and polyethylene oxide solution as pore-forming agent, and the effect of porous hydrogel scaffolds on cellular behavior. METHODS: Methacrylate anhydride gelatin hydrogel solution and polyethylene oxide solution were prepared, and mixed in a volume ratio of 4:1, 3:1, 2:1, and 1:1, separately. After adding bone marrow mesenchymal stem cells, a three-dimensional porous hydrogel scaffold that could carry cells were designed and constructed to characterize the microscopic morphology and mechanical properties of the scaffold. The live/dead staining was utilized to observe cell compatibility, and cytoskeleton staining was applied to observe cell morphology. RESULTS AND CONCLUSION: (1) The scanning electron microscope showed that the cross-sections of scaffolds of each group displayed uniformly distributed porous structures, and the pores were connected to each other. The pores were oval-like or round-like. As the proportion of polyethylene oxide solution increased, the pore size of the scaffolds and porosity increased. (2) Multi-frequency strain curves showed that the strain of scaffolds of each group could reach more than 40%, among which the strain of the scaffold with a volume ratio of 1:1 could reach 60%. As the proportion of polyethylene oxide solution increased, the storage modulus of the scaffold decreased, and the mechanical strength of the volume ratio 1:1 scaffold was poor, and it could barely maintain the shape of the scaffold. (3) The live/dead staining of 14 days in vitro culture showed that the survival rate of cells of scaffolds in each group was >85%. (4) Staining of the cytoskeleton cultured for 14 days in vitro showed that most of the cells in the scaffold with a volume ratio of 3:1 were short spindle-shaped, and the cells in the scaffold with a volume ratio of 2:1 and 1:1 were star-shaped or long spindle-shaped. A wide range of connections was established among cells. (5) Three-dimensional cell-carrying porous hydrogel scaffold can significantly promote the spreading and proliferation of bone marrow mesenchymal stem cells. Porous hydrogels with a volume ratio of 2:1 have both mechanical strength and biological properties and can be a good platform for three-dimensional cell culture. Key words:bone defect, porous hydrogel, methacrylate anhydride gelatin, tissue engineering, scaffold, bone marrow mesenchymal stem cells, three-dimensional culture, light curing |