Effects of dynamic pressure on the proliferation and mechanical properties of rabbit adipose mesenchymal stem cells transfected with insulin-like growth factor-1
10.3969/j.issn.2095-4344.3514
- Author:
Zhang CHUANHUI
1
Author Information
1. Department of Orthopedics, Chaoyang Central Hospital
- Publication Type:Journal Article
- Keywords:
Adipose mesenchymal stem cells;
Chondrocytes;
Elasticity modulus;
Insulin-like growth factor-1;
Pressure;
Stem cells;
Tissue engineering
- From:
Chinese Journal of Tissue Engineering Research
2021;25(13):1982-1987
- CountryChina
- Language:Chinese
-
Abstract:
BACKGROUND: Adipose mesenchymal stem cells are currently recognized as excellent seed cells for tissue engineering cartilage. Gene transfection technology can effectively induce them to differentiate into cartilage. The bioreactor is used to simulate the mechanical environment in vivo. It is a new idea for the majority of scholars to explore the construction of tissue engineering cartilage in vitro. OBJECTIVE: To investigate the effects of cyclic dynamic compressive stress combined with insulin-like growth factor-1 gene transfection on the proliferation and elastic modulus of rabbit adipose mesenchymal stem cells implanted in chitosan/gelatin scaffold. METHODS: Rabbit adipose mesenchymal stem cells were transfected with pcDNA3.1-IGF-1 gene mediated by liposome. The stable transfected cell lines were screened by G418. The adipose mesenchymal stem cells transfected with or without insulin-like growth factor-1 gene were inoculated in chitosan/ gelatin scaffold at the density of 5×1010 L-1 for 2 days, and cultured under dynamic pressure (2% at 1 Hz, 4 hours per day) or static culture conditions for 7 days, respectively. The morphological changes of the cell/scaffold complex were observed by scanning electron microscope, Masson trichrome staining and alcian blue staining. The cell proliferation curve was drawn by MTT assay. The cell proliferation efficiency and distribution were evaluated by CM-Dil fluorescence-labeling method, and the content of total glycosaminoglycan was quantitatively determined by DMMB. The differences of type II collagen among different groups were compared with real time PCR. Compressive mechanical properties of the cell/scaffold constructs were assessed using a BioDynamic™ mechanical tester, and the corresponding elastic modulus was calculated. RESULTS AND CONCLUSION: Dynamic pressure combined with insulin-like growth factor-1 transfection could significantly improve the cell proliferation ability of the cell/scaffold complex; the cell distribution was more uniform; glycosaminoglycan and collagen secretion in the cartilage-specific extracellular matrix were increased; the expression levels of type II collagen were up-regulated; and the mechanical properties were significantly improved. The cell proliferation and elastic modulus of insulin-like growth factor-1 group were better than those of single pressure group, but the distribution of cells in scaffolds was more uniform under dynamic pressure. The results indicate that both dynamic pressure and insulin-like growth factor-1 gene transfection can significantly improve the proliferation and mechanical properties of rabbit adipose mesenchymal stem cells; the two have synergistic effect.
