Fabrication and biocompatibility of injectable gelatin-methacryloyl/cartilage-derived matrix particles composite hydrogel scaffold

doi:10.12307/2022.244

Abstract

Abstract: BACKGROUND: Articular cartilage regeneration and repair is still a challenging medical problem. The injectable hydrogel scaffold that constructs a biomimetic microenvironment for cell growth is of great significance for the regeneration of articular cartilage.
OBJECTIVE: To prepare injectable gelatin-methacryloyl/cartilage-derived matrix particles (GelMA/CDMPs) composite hydrogel scaffold, and evaluate its biocompatibility.
METHODS: (1) CDMPs were prepared by physical pulverization and chemical enzymatic methods, and GelMA was prepared by the reaction of methacrylic anhydride grafted gelatin. The two were used as raw materials to prepare five different GelMA/CDMP composite hydrogel scaffolds (where the CDMP mass concentrations were 0, 5, 10, 20, and 30 g/L), and the mass concentration of CDMPs with the best mechanical properties was determined through mechanical tests. (2) Rabbit bone marrow mesenchymal stem cells were encapsulated in the GelMA/CDMPs composite hydrogel scaffolds (as the experimental group); bone marrow mesenchymal stem cells were encapsulated in GelMA hydrogel scaffolds (as the control group); both of which were cultured in the chondrogenic medium. The cell viability and cell proliferation of the two groups of stents were evaluated through cell live/dead staining and CCK-8 cell proliferation experiments. The morphology and spreading ability of cells in the scaffolds were observed by cytoskeleton fluorescence staining. The secretion of cartilage matrix of bone marrow mesenchymal stem cells in the scaffolds was evaluated by histological staining and quantitative glycosaminoglycan test.
RESULTS AND CONCLUSION: (1) Mechanical tests showed that compressive modulus of GelMA/CDMPs 10 g/L composite hydrogel scaffolds was significantly higher than that of other groups (P < 0.05). And this concentration was selected for the subsequent experiments. (2) The cell live/dead staining demonstrated that cell viability of bone marrow mesenchymal stem cells in the two groups of scaffolds was above 90% after 1 and 5 days of culture. After 3- and 5-day culture, the cell proliferation of the experimental group was faster than that of the control group (P < 0.05). (3) Fluorescence staining of the cytoskeleton after culture for 7 days showed that the cell morphology in the control group was spherical; and only a small part of the cells formed filopodia. The cell morphology in the experimental group was stretched and showed irregular polygons. (4) After 21 days of culture, histological and immunohistochemical staining showed that the cell morphology of bone marrow mesenchymal stem cells in the two groups of scaffolds was similar to chondrocytes morphology; and more cartilage matrix was secreted by bone marrow mesenchymal stem cells in the experimental group. (5) After 7- and 21-day culture, the content of glycosaminoglycan was significantly higher in the experimental group than that in the control group (P < 0.05). (6) These results suggest that GelMA/CDMPs composite hydrogel scaffold can better mimic the microenvironment of chondrocyte growth, and then promote the growth, proliferation and chondrogenesis of bone marrow mesenchymal stem cells.

Key words: cartilage, material, tissue engineering, cartilage-derived matrix, hydrogel, GelMA, injectability, mesenchymal stem cell, biocompatibility

CLC Number:

Huang Bo, Chen Mingxue, Peng Liqing, Luo Xujiang, Li Huo, Wang Hao, Tian Qinyu, Lu Xiaobo, Liu Shuyun, Guo Quanyi . Fabrication and biocompatibility of injectable gelatin-methacryloyl/cartilage-derived matrix particles composite hydrogel scaffold[J]. Chinese Journal of Tissue Engineering Research, 2022, 26(16): 2480-2486.

Figures/Tables 8

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