Construction of tissue engineered bone by poly(lactic-co-glycolic acid) scaffold carrying human bone marrow mesenchymal stem cells

doi:10.3969/j.issn.2095-4344.1347

Abstract

Abstract:

BACKGROUND: Poly(lactic-co-glycolic acid) (PLGA) is one of the few biomaterials approved for experimental and clinical use. It has good biocompatibility, biodegradability and excellent mechanical properties, and is a good scaffold carrier in tissue engineering.

OBJECTIVE: To investigate the in vitro biocompatibility and in vivo ectopic osteogenic differentiation of PLGA nano-scaffold material carrying human bone marrow mesenchymal stem cells.

METHODS: 150 g/L PLGA solution was electrospun into fiber membrane by electrospinning technique. Passage 3 human bone marrow mesenchymal stem cells (harvesting of human bone marrow sample was approved by Ethics Committee of Affiliated Hospital of Guizhou Medical University, China) were seeded into PLGA nano-scaffold and cultured with osteogenic induction medium for 7 days. After 4',6-diamidino-2-phenylindole staining and acridine orange fluorescent staining, the growth of cells/scaffold complex was observed under scanning electron microscope. Passage 3 human bone marrow mesenchymal stem cells were seeded on PLGA nano-scaffold and cultured with osteogenic induction medium (experimental group) or without (control group). After 14 days of culture, cell-scaffold complex was subcutaneously implanted into the nude mice. At 12 weeks after implantation, the cell-scaffold complex was removed and stained with hematoxylin-eosin, alizarin red, alkaline phosphatase, and collagen I immunohistochemical staining to observe ectopic osteogenesis.

RESULTS AND CONCLUSION: 4',6-Diamidino-2-phenylindole staining and acridine orange fluorescent staining revealed that human bone marrow mesenchymal stem cells adhered well on the surface of the scaffold. Scanning electron microscopy showed that after osteogenic induction, human bone marrow mesenchymal stem cells grew well on PLGA scaffold, and cells produced abundant extracellular matrix. At 12 weeks after implantation, the cells were positive for alizarin red staining, alkaline phosphatase staining and collagen type I immunohistochemical staining. Positive expression in the experimental group was stronger than that in the control group. Hematoxylin-eosin staining showed numerous osteoblasts and typical bone lacunaes in the experimental group and a small amount of bone tissue in the control group. These results suggest that PLGA nano-scaffold material exhibits good biocompatibility and in vivo ectopic osteogenic capacity, and can be used as scaffold material in bone tissue engineering.

Key words: poly(lactic-co-glycolic acid), PLGA, nanofiber membrane, electrospinning, scaffold, human bone marrow mesenchymal stem cells, biocompatibility, tissue engineering, ectopic osteogenesis

CLC Number:

Chen Jiao, Shu Liping, Li Xuanze, Liu Qin, Wu Ying, Liu Yin, Wang Weiyu, Liu Jun, Ye Chuan, Ma Minxian. Construction of tissue engineered bone by poly(lactic-co-glycolic acid) scaffold carrying human bone marrow mesenchymal stem cells[J]. Chinese Journal of Tissue Engineering Research, 2019, 23(26): 4109-4114.

Figures/Tables 5

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