国际口腔医学杂志

国际口腔医学杂志 ›› 2020, Vol. 47 ›› Issue (3): 278-285.doi: 10.7518/gjkq.2020061

• 论著 • 上一篇    下一篇

低温氩氧等离子体处理的无机牛骨对MC3T3-E1细胞黏附、增殖及分化的影响

马凯,李昊,赵红梅,王永亮,刘杰,柏娜()   

  1. 青岛大学附属医院口腔修复科 青岛大学口腔医学院 青岛 266003
  • 收稿日期:2019-10-22 修回日期:2020-01-06 出版日期:2020-05-01 发布日期:2020-05-08
  • 通讯作者: 柏娜
  • 作者简介:马凯,住院医师,硕士,Email：18661899317@163.com
  • 基金资助:
    国家自然科学基金(81500882)

Effects of inorganic bovine bone treated with low temperature argon-oxygen plasma on the adhesion, proliferation, and differentiation of MC3T3-E1 cells

Ma Kai,Li Hao,Zhao Hongmei,Wang Yongliang,Liu Jie,Bai Na()   

  1. Dept. of Prosthodontics, The Affiliated Hospital of Qingdao University, School of Stomatology of Qingdao University, Qingdao 266003, China
  • Received:2019-10-22 Revised:2020-01-06 Online:2020-05-01 Published:2020-05-08
  • Contact: Na Bai
  • Supported by:
    National Natural Science Foundation of China(81500882)

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摘要：

目的 研究小鼠胚胎成骨细胞MC3T3-E1在低温氩氧等离子体处理的无机牛骨上的黏附、增殖及分化特征。方法 使用低温氩氧等离子体对无机牛骨进行表面活化（实验组）后,使用扫描电子显微镜（SEM）观察无机牛骨表面形貌的变化,X射线光电子能谱分析（XPS）检测表面元素的组成。将MC3T3-E1细胞接种于低温氩氧等离子体处理的无机牛骨表面,使用SEM观察细胞的黏附形态,CCK-8法检测细胞1、3、5 d的增殖变化,碱性磷酸酶（ALP）法检测细胞7、14 d的分化状态。以不作处理的无机牛骨作为对照。结果 对照组与实验组的表面形貌无明显改变;在表面材料的元素组成上,实验组表面碳元素减少,氧、钙、磷元素均增高。实验组MC3T3-E1细胞的黏附更充分,细胞伸出伪足;培养1、3、5 d时,实验组细胞增殖数量明显高于对照组;培养14 d时,实验组ALP活性明显高于对照组（P<0.05）。结论 低温氩氧等离子体处理对无机牛骨表面小鼠胚胎成骨细胞的黏附、增殖及分化具有一定的促进作用。

关键词: 低温等离子体, 无机牛骨, 小鼠胚胎成骨细胞MC3T3-E1, 引导骨组织再生

Abstract:

Objective To study the adhesion, proliferation, and differentiation of mouse embryonic osteoblast MC3T3-E1 on inorganic bovine bone treated with low temperature argon-oxygen plasma. Methods After the surface activation of inorganic bovine bone by low temperature argon-oxygen plasma, the surface morphology of inorganic bovine bone before and after treatment was observed by scanning electron microscopy (SEM), and the surface elemental composition was detected by X-ray photoelectron spectroscopy. The MC3T3-E1 cells were inoculated on the surface of inorganic bovine bone treated with low temperature argon-oxygen plasma, and the cell adhesion morphology was observed by SEM. The proliferation of cells on days 1, 3, and 5 was detected using the CCK-8 method, and the differentiation of cells on days 7 and 14 was detected using the alkaline phosphatase (ALP) method. The control group comprised the inorganic bovine bone without treatment. Results No significant change was observed in the surface morphology of the inorganic bovine bone of the control and the argon-oxygen groups. Relative to the composition of the material elements in the control group, the argon-oxygen group had decreased surface C element and increased O, Ca, and P elements. Under SEM, the cell adhesion of the argon-oxygen group was more complete, and the cell extended pseudopodia. The cell proliferation test results showed that the number of cell proliferation in the argon-oxygen group was significantly higher than that in the control group at days 1, 3, and 5. The cell differentiation results showed that the ALP activity in the argon-oxygen group was higher than that in control group at day 14. Conclusion The inorganic bovine bone treated with low temperature argon-oxygen plasma can promote the adhesion, proliferation, and differentiation of mouse embryonic osteoblast cells.

Key words: low temperature plasma, inorganic bovine bone, mouse embryonic osteoblast MC3T3-E1, guided bone regeneration

中图分类号: 

  • Q25

图 1

低温氩氧等离子体系统原理示意图"

图 2

低温氩氧等离子体系统处理时间与材料表面温度的关系"

图 3

温度过高导致骨块内部连接结构断裂 SEM × 2 000"

图 4

实验组和对照组的表面形貌 SEM A：对照组,× 2 000;B：对照组,×10 000;C：实验组,× 2 000;D：实验组,× 10 000。"

表 1

两组骨块表面元素百分比的XPS分析"

组别 C1s O1s P2p Ca2p
对照 50.92 37.05 5.94 6.10
实验 43.09 43.11 6.72 7.08

图 5

XPS检测两组骨块表面元素的变化 A：碳（C）元素;B：氧（O）元素;C：钙（Ca）元素;D：磷（P）元素;E：全谱变化;F：对照组与实验组各元素的百分比。"

图 6

实验组即刻及暴露2 d后表面各元素百分比"

图 7

对照组和实验组表面MC3T3-E1细胞形态 SEM A：对照组,× 2 500;B：对照组,× 5 000;C：实验组,× 2 500;D：实验组,× 5 000。"

图 8

实验组和对照组表面成骨细胞的黏附面积百分比 ??P<0.01。"

图 9

MC3T3-E1细胞增殖结果 ?P<0.05,??P<0.01。"

图 10

MC3T3-E1细胞ALP活性检测结果 ?P<0.05。"

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