Chinese Journal of Tissue Engineering Research ›› 2020, Vol. 24 ›› Issue (1): 130-135.doi: 10.3969/j.issn.2095-4344.1845

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Effect of sex combing protein 1 on proliferation and differentiation of osteoblasts in inflammatory microenvironment#br#

Xu Huijun, Shi Dongmei, Zhang Mi, Wu Saixuan, Dong Ming, Lu Ying, Niu Weidong   

  1. Department of Endodontics, School of Stomatology, Dalian Medical University, Dalian 116044, Liaoning Province, China
  • Received:2019-02-25 Revised:2019-03-05 Accepted:2019-04-19 Online:2020-01-08 Published:2019-12-13
  • Contact: Niu Weidong, MD, Professor, Doctoral supervisor, Department of Endodontics, School of Stomatology, Dalian Medical University, Dalian 116044, Liaoning Province, China
  • About author:Xu Huijun, Master candidate, Physician, Department of Endodontics, School of Stomatology, Dalian Medical University, Dalian 116044, Liaoning Province, China Shi Dongmei, Master candidate, Physician, Department of Endodontics, School of Stomatology, Dalian Medical University, Dalian 116044, Liaoning Province, China Xu Huijun and Shi Dongmei contributed equally to this work
  • Supported by:
    the National Natural Science Foundation of China (Youth Project), No. 81700962

Abstract:

BACKGROUND: Studies have shown that the loss of sex combing protein 1 (Asxl1) can lead to the occurrence of bone dysplasia and bone defects, but the relationship between this factor and bone destruction in the microenvironment of apical periodontitis has not been reported.

OBJECTIVE: To study the effect of Asxl1 on proliferation and differentiation of osteoblasts in an inflammatory microenvironment.

METHODS: MC3T3-E1 cells were excited by lipopolysaccharide to establish an in vitro inflammatory microenvironment. The best concentration and optimal action time of lipopolysaccharide were screened by cell counting kit-8 test. MC3T3-E1 cells were then stimulated with 20 mg/L lipopolysaccharide for 24 hours. The expression levels of Asxl1 protein and mRNA were detected by immunofluorescence and real-time PCR, respectively. After lipopolysaccharide stimulated the formation of inflammatory microenvironment, Asxl1-Si RNA was transfected for 24 hours, cell counting kit-8 was applied to detect the activity of cell proliferation, and real-time PCR was used to detect the expression levels of Asxl1 and osteogenic related genes ALP and RUNX2 mRNA.

RESULTS AND CONCLUSION: After lipopolysaccharides stimulation of MC3T3-E1 cells, the expression levels of Asxl1 protein and mRNA were decreased. Under the inflammatory microenvironment, the proliferation activity of MC3T3-E1 cells transfected with Asxl1-Si RNA for 24 hours was significantly decreased, and the expression levels of Asxl1, ALP and RUNX2 mRNA were markedly decreased. These findings indicate that Asxl1 may influence the proliferation and differentiation of osteoblasts by involvement in the process of inflammatory reaction, thereby participating in bone destruction.

Key words: MC3T3-E1 cells, Asxl1, periapical periodontitis, bone destruction, gene transfection, lipopolysaccharide, osteogenic differentiation, National Natural Science Foundation of China

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