中国实用口腔科杂志

中国实用口腔科杂志

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不同质量分数的钛-铜合金对MC3T3-E1细胞分化的影响研究

马凯1，王敏雅1，李悦2，张燕婷1，李胜义3，张二林3，赵宝红1，邓春富1   

  1. 1. 中国医科大学口腔医学院·附属口腔医院种植中心，辽宁省口腔医学研究所口腔种植研究室，辽宁  沈阳110002；2. 沈阳市和平区新兴齿科门诊部，辽宁  沈阳110005；3. 东北大学材料各向异性与织构教育部重点实验室，辽宁  沈阳  110819
  • 出版日期:2018-06-15 发布日期:2018-07-10
  • 基金资助:

    国家自然科学基金（51775096）；辽宁省自然科学基金（20170541059）；辽宁省科技计划项目（2013225049）

  • Online:2018-06-15 Published:2018-07-10

摘要：

目的　探究不同质量分数的钛-铜合金对MC3T3-E1细胞分化的影响，并寻找促进细胞分化作用最佳的含铜质量分数。方法　将不同质量分数的钛-铜合金样片分组，设组1（cp Ti）为对照组，组2（Ti-2wt%Cu）、组3（Ti-5wt%Cu）、组4（Ti-10wt%Cu）为实验组。将MC3T3-E1细胞按2 × 104/孔的密度接种到包埋样片的24孔培养板中行共培养。在第3、5、10和15 d时，用微量酶标法检测成骨细胞内碱性磷酸酶（ALP）活性表达情况；在第7、14和21 d时，分别用酶联免疫吸附试验（ELISA）法检测上清液中Ⅰ型胶原蛋白（COL-Ⅰ）、骨钙素（OC）活性表达情况。采用SPSS 20.0统计分析软件包对所有检测数据进行单因素方差分析，比较各组间差异。 结果　Ti-2wt%Cu最有利于ALP活性表达（P＜0.05），共培养第10 d时达最大值；Ti-5wt%Cu最有利于COL-Ⅰ活性表达（P＜0.05），共培养第7 d时达最大值；Ti-5wt%Cu最有利于OC活性表达（P＜0.05），共培养第14 d时达最大值。 结论　与cp Ti相比较，低质量分数钛-铜（Ti-2wt%Cu、Ti-5wt%Cu）对MC3T3-E1细胞分化有促进作用，表现为时间依赖性；高质量分数钛-铜（Ti-10wt%Cu）对MC3T3-E1细胞分化有抑制作用。相对而言，Ti-5wt%Cu合金对MC3T3-E1细胞分化作用最佳。

关键词: 钛-铜合金, 铜含量, MC3T3-E1细胞, 细胞分化

Abstract:

Objective　To explore the effect of different concentration of titanium-copper alloy on the differentiation of MC3T3-E1 cells and to find the best mass fraction of copper promoting cell differentiation. Methods　Different concentrations of titanium-copper alloy were divided into groups. Mark cp Ti with Group1，as control group；mark Ti-2 wt% Cu with Group 2，Ti-5wt% Cu with Group 3，and Ti-10 wt% Cu with Group 4，as experiment groups. The MC3T3-E1 cells were inoculated into a 24-well culture plates with prepared samples for co-culture at a density of 2×104 per well. The expression of ALP activity of osteoblasts was detected by micro-enzyme-labeled method at 3 d，5 d，10 d and 15 d. The expression of type I collagen and osteocalcin in the supernatant of the co-cultured osteoblasts were detected by ELISA at 7 d，14 d and 21 d. All data were analyzed by means of  one-way ANOVA with SPSS 20.0 software package. Results　Ti-2wt% Cu was the most favorable for the expression of alkaline phosphatase activity （P＜0.05）and reached the maximum at the 10th day after co-culture. Ti-5wt% Cu was the most favorable for the expression of type I collagen protein（P＜0.05）and reached the maximum at the 7th day after co-culture. Ti-5wt% Cu was the most favorable for the expression of osteocalcin（P＜0.05） and reached the maximum at the 14th day. Conclusion　Compared with cp Ti，low concentration of Ti-Cu （2wt%，5wt% ） alloy promotes the differentiation of MC3T3-E1 cells，being time-dependent，and high concentration of Ti-Cu （10wt%） alloy inhibits the differentiation of MC3T3-E1cells. Relatively speaking，Ti- 5wt%Cu alloy promotes the differentiation of MC3T3-E1 cells best.

Key words: titanium-copper alloy, copper concentration, MC3T3-E1 cell, cell differentiation