Chinese Journal of Tissue Engineering Research ›› 2019, Vol. 23 ›› Issue (18): 2888-2893.doi: 10.3969/j.issn.2095-4344.1713

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Osteogenic differentiation of MC3T3-E1 cells induced by osteopractic total flavone and nano-hydroxyapatite/collagen composite and the underlying mechanism

Li Jinyu1, Yu Xing1, Jiang Junjie2, Xu Lin1, Zhao Xueqian1, Sun Qi1, Zheng Chenying1, Bai Chunxiao1, Liu Chuyin1, Zhang Zhe1, Jia Yusong1 
  

  1. 1Department of Orthopedics, Dongzhimen Hospital, Beijing University of Chinese Medicine, Beijing 100700, China; 2Institute for Basic Research in Clinical Medicine, China Academy of Chinese Medical Sciences, Beijing 100700, China
  • Received:2019-01-22 Online:2019-06-28 Published:2019-06-28
  • Contact: Jia Yusong, Chief physician, Department of Orthopedics, Dongzhimen Hospital, Beijing University of Chinese Medicine, Beijing 100700, China
  • About author:Li Jinyu, MD, Associate chief physician, Department of Orthopedics, Dongzhimen Hospital, Beijing University of Chinese Medicine, Beijing 100700, China
  • Supported by:

    the National Natural Science Foundation of China, No. 81503601 (to LJY) and 81603517 (to JJJ); the Young Talent Project of Dongzhimen Hospital of Beijing University of Chinese Medicine, No. DZMYS-201802 (to LJY)

Abstract:

BACKGROUND: Preliminary study has found that osteopractic total flavone can promote osteogenic differentiation of MC3T3-E1 cells on the surface of nano-hydroxyapatite/collagen composite, but the underlying mechanism needs to be studied in depth.
OBJECTIVE: To investigate the effect of osteopractic total flavone combined with nano-hydroxyapatite/collagen composite on the osteogenic differentiation of MC3T3-E1 cells and its mechanism.
METHODS: MC3T3-E1 cells were seeded on the surface of nano-hydroxyapatite/collagen composite and cultured in culture media containing 0, 100 and 250 mg/L osteopractic total flavone, respectively. After 1, 3, 5, 7, 9, 11 and 14 days of culture, the activity of alkaline phosphatase was detected. After 1, 2, 3 and 4 weeks of culture, the level of osteocalcin in the culture supernatant was detected by ELISA. Alizarin red staining was used to observe the formation of calcified nodules at 7, 14 and 21 days of culture. At 14 days of culture, the expression of osteogenic related genes was detected by PCR.
RESULTS AND CONCLUSION: (1) There was no significant difference in the alkaline phosphatase activity at 1, 3 and 14 days among groups (P > 0.05). At 5, 7 and 9 days of culture, the alkaline phosphatase activity in the 100 and 250 mg/L groups was higher than that in the 0 mg/L group (P < 0.05), and there was no significant difference between 100 and 250 mg/L groups (P > 0.05). (2) The mass concentration of osteocalcin in the 100 and 250 mg/L groups was higher than that in the 0 mg/L group at 1, 2, 3 and 4 weeks of culture (P < 0.05), and there was no significant difference between 100 and 250 mg/L groups (P > 0.05). (3) At 7, 14 and 21 days of culture, obvious calcified nodules were visible in the 100 and 250 mg/L groups. (4) At 14 days of culture, the mRNA expression levels of osteocalcin, type I collagen and osteopontin in the 100 and 250 mg/L groups were higher than that in the 0 mg/L group (P < 0.05), and there was no significant difference between 100 and 250 mg/L groups (P > 0.05). (5) These results imply that osteopractic total flavone combined with nano-hydroxyapatite/collagen composite can induce MC3T3-E1 cells to differentiate into osteoblasts, which may be by increasing alkaline phosphatase activity, osteocalcin level and mRNA expression of type I collagen, osteocalcin and osteopontin.

Key words: nano-hydroxyapatite/collagen composite, osteopractic total flauone, MC3T3-E1 cells, osteogenic differentiation, osteopontin, osteocalcin, the National Natural Science Foundation of China

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