Int J Stomatol

Inter J Stomatol ›› 2019, Vol. 46 ›› Issue (2): 203-208.doi: 10.7518/gjkq.2019025

• Original Articles • Previous Articles     Next Articles

Effect of the titanium surface immobilised by arginyl-glycyl-aspartate peptide self-assembling multilayers on mouse osteoblast-like cell MC3T3-E1

Fei Liu1,Yuntao Zhang2(),Xiangrui Ma3,Yajie Zhang1,Yunhao Wang1   

  1. 1. School of Stomatology, Binzhou Medical University, Binzhou 256600, China
    2.Dept. of Prosthodontics, Affiliated Hospital of Binzhou Medical University, Binzhou 256600, China
    3.Dept. of Oral and Maxillofacial Surgery, Affiliated Hospital of Binzhou Medical University, Binzhou 256600, China)
  • Received:2018-07-04 Revised:2018-10-11 Online:2019-03-01 Published:2019-03-15
  • Contact: Yuntao Zhang E-mail:zytao999@163.com
  • Supported by:
    This study was supported by Projects of Medical and Health Technology Development Program of Shandong Province(2017WS231)

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Abstract:

Objective This study aimed to investigate the effect of the titanium surface immobilised by arginyl-glycyl-aspartate (RGD) peptide self-assembling multilayers on the mouse osteoblast-like cell MC3T3-E1. Methods Metal specimens were randomly divided into three groups: pure titanium (PT) group, NaOH-treated titanium (NT) group and RGD-modified titanium (RT) group. The morphology of the specimens was inspected by scanning electron microscopy (SEM). MC3T3-E1 cells were cultured on the surface of specimens. The adhesion and proliferation of MC3T3-E1 cells were observed by SEM and 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) assay. The mRNA expression levels of osteocalcin and osteoprotegerin were quantified by real-time quantitative polymerase chain reaction. Results The sample surface of the PT group showed uniform and clear scratches, whereas that of the NT group showed microporous structure. Meanwhile, the surface structure of the RT group resembled the small bulges to a web surface. Cells adhering on RT disks exhibited active spreading and polygonous shape, with more filopodia. MTT assay showed that the adhesion rate in the RT group was the highest among the four groups; no considerable difference existed between the control group (withont titanium) and PT group. The cell proliferation rate of RT group was also the highest (P<0.05), especially at 1 and 3 days. In the RT group, osteocalcin mRNA and osteoprotegerin mRNA expression levels decreased more than those in the control group and PT group at 14 days (P<0.05). Conclusion RT can promote cell adhesion and proliferation, and increase osteocalcin mRNA and osteoprotegerin mRNA expression levels in MC3T3 cells.

Key words: layer-by-layer self-assembly, arginyl-glycyl-aspartate peptide, mouse osteoblast-like cell MC3T3-E1

CLC Number: 

  • Q813.1 +1

TrendMD: 

Fig 1

Surface topography of the titanium plates SEM × 5 000"

Tab 1

The effect of different titanium plates on the adhesion of MC3T3-E1"

分组 1 h 4 h 12 h 24 h
对照组 0.035±0.006 0.044±0.006 0.092±0.006 0.129±0.004
PT组 0.036±0.005 0.045±0.005 0.095±0.005 0.134±0.005
NT组 0.028±0.005*# 0.035±0.005*# 0.075±0.004*# 0.100±0.009*#
RT组 0.049±0.003*#& 0.064±0.005*#& 0.104±0.005*#& 0.153±0.005*#&

Fig 2

Adhesion morphology of MC3T3 on surface of titanium plates SEM × 1 000"

Tab 2

The effect of different titanium plates on the proliferation of MC3T3-E1"

分组 1 d 3 d 5 d 7 d
对照组 0.128±0.004 0.375±0.069 0.931±0.109 1.115±0.142
PT组 0.131±0.010 0.386±0.027 0.949±0.115 1.064±0.137
NT组 0.109±0.014*# 0.154±0.041*# 0.286±0.048*# 0.679±0.060*#
RT组 0.155±0.009*#& 0.544±0.118*#& 0.969±0.166& 1.138±0.082&

Fig 3

mRNA expression of osteocalcin and osteoprotegerin in MC3T3-E1 on different titanium plates at different times"

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