钴铬钼合金对L929细胞生物学行为的影响

上海口腔医学 ›› 2013, Vol. 22 ›› Issue (4): 407-413.

钴铬钼合金对L929细胞生物学行为的影响

周群¹，苗磊¹，冯靖雯²，余和东¹，郭蕾¹，林晓萍¹

(1.中国医科大学附属盛京医院口腔科，辽宁沈阳 110004；2.中国医科大学附属口腔医院正畸科，辽宁沈阳 110001)

收稿日期:2013-01-14 修回日期:2013-03-08 出版日期:2013-08-10 发布日期:2013-08-10
通讯作者: 周群，E-mail: 263251815@qq.com
作者简介:周群(1985-)，男，博士，副教授

The influence of Co-Cr-Mo alloys on biological behaviour of L929 mouse fibroblasts

ZHOU Qun¹, MIAO Lei¹,FENG Jing-wen², YU He-dong¹, GUO Lei¹, LIN Xiao-ping¹

1.Department of Prosthodontics, Shengjing Hospital of China Medical University. Shenyang 110004; 2.Department of Orthodontics, Stomatological School of China Medical University. Shenyang 110002, Liaoning Province, China

Received:2013-01-14 Revised:2013-03-08 Online:2013-08-10 Published:2013-08-10

摘要/Abstract

摘要： 目的：研究牙科用钴铬钼合金对L929细胞生物学行为的影响。方法：制备医用纯钛、钴铬钼合金、钴铬合金、镍铬合金的浸提液，在浸提液与L929细胞作用24 、48 、72 h后，用倒置相差显微镜观察细胞的生长情况，并通过CCK-8实验评定4种材料的细胞毒性；采用流式细胞仪Annexin V-FITC/PI 双染法评价材料对细胞凋亡的影响；应用吖啶橙染色法检测L929细胞在材料表面的黏附情况；采用SPSS17.0软件包对数据进行统计学分析。结果：倒置相差显微镜下观察，各时间点除镍铬合金组可见少量细胞核固缩，呈现轻微中毒状态外，其余3组细胞生长情况良好；钴铬钼合金组在各时间点的OD值、细胞凋亡率、细胞黏附数量均显著低于医用纯钛组(P<0.05)，而高于钴铬合金组 (P<0.05)和镍铬合金组(P<0.05)；观察期内，医用纯钛组、钴铬钼合金组、钴铬合金组细胞毒性均为1级，镍铬合金组为2级，表现为轻微毒性。结论：钴铬钼合金对L929细胞的生物学行为无明显不良影响，符合口腔生物材料的临床应用要求，具有良好的生物相容性。

关键词: 钴铬钼合金, 细胞毒性, L929细胞

Abstract: PURPOSE: To investigate the influence of dental Co-Cr-Mo alloys on biological behaviour of L929 mouse fibroblasts. METHODS: Leaching liquor of medical pure titanium, Co-Cr-Mo alloys, Co-Cr alloys and Ni-Cr alloys was prepared and used to culture the L929 cells for 24 h, 48 h and 72 h, respectively; then the growth of L929 cells was observed under inverted phase contrast microscope, and the cytotoxicity grades of 4 kinds of materials were evaluated by using CCK-8 test. The apoptosis of L929 cells was measured by flow cytometry(FCM) and acridine orange staining was used to observe the L929 cells adhered on the surface of samples under fluorescent microscope. The data was statistically analyzed with SPSS17.0 software package. RESULTS: Under inverted phase contrast microscope, at each time point, L929 cells grew in a good condition except in the Ni-Cr alloys group that minor karyopyknosis was indicating slight celluar cytotoxicity. Optical density (OD), apoptosis rate and cell adhesion number in Co-Cr-Mo alloys group at each time point were significantly less than in medical pure titanium group (P<0.05), and higher than in Co-Cr alloys group (P<0.05) and Ni-Cr alloys group (P<0.05). During observation period, the cytotoxicity grades of medical pure titanium group, Co-Cr-Mo alloys group and Co-Cr alloys group was grade 1, while Ni-Cr alloy group was grade 2, i.e. mild cytotoxicity. CONCLUSIONS: Co-Cr-Mo alloys have no adverse effect on the biological behavior of L929 cells, which may be suitable for dental clinical application with good biocompatibility.

Key words: Co-Cr-Mo alloy, Cytotoxicity, L929 cells

中图分类号:

R783.1

周群, 苗磊, 冯靖雯, 余和东, 郭蕾, 林晓萍. 钴铬钼合金对L929细胞生物学行为的影响[J]. 上海口腔医学, 2013, 22(4): 407-413.

ZHOU Qun, MIAO Lei, FENG Jing-wen, YU He-dong, GUO Lei, LIN Xiao-ping. The influence of Co-Cr-Mo alloys on biological behaviour of L929 mouse fibroblasts[J]. Shanghai Journal of Stomatology, 2013, 22(4): 407-413.

参考文献

[1] Reibel J. Prognosis of oral pre-malignant lesions: significance of clinical, histopathological, and molecular biological characteristics[J]. Crit Rev Oral Biol Med, 2003, 14(1): 47-62.
[2] Nganbe M, Khan U, Louati H, et a1. In vitro assessment of strength, fatigue durability, and disassembly of Ti6Al4V and CoCrMo necks in modular total hip replacements[J]. J Biomed Mater Res B Appl Biomater, 2011, 97(1): 132-138.
[3] 孙凤. Vitallium2000铸造支架及分裂基托设计的临床应用[J]. 华西口腔医学杂志, 2009, 27(2): 119-121, 125.
[4] Garcia Lda F, Marques AA, Roselino Lde M, et al. Biocompatibility evaluation of Epiphany/Resilon root canal filling system in subcutaneous tissue of rats[J]. J Endod, 2010, 36(1): 110-114.
[5] 张德贵.整体铸造网状支架混合支持式义齿临床设计与观察[J]. 口腔医学, 2004, 24(1): 63-64.
[6] Larsen A, Stoltenberg M, Danscher G. In vitro liberation of charged gold atoms: autometallographic tracing of gold ions released by macrophages grown on metallic gold surfaces[J]. Histochem Cell Biol, 2007, 128(1): 1-6.
[7] 叶荣荣, 沈庆平, 乔广艳. 3种临床常用冠桥材料对体外培养鼠成纤维细胞毒性的比较[J]. 上海口腔医学, 2008, 17(3): 308-312.
[8] Ohkubo C, Hanatani S, Hosoi T. Present status of titanium removable dentures-a review of the literature[J]. J Oral Rehabil, 2008, 35(9): 706-714.
[9] Lü X, Bao X, Huang Y, et al. Mechanisms of cytotoxicity of nickel ions based on gene expression profiles[J]. Biomaterials, 2009, 30(2): 141-148.
[10] 胡滨,陈洁,张富强. 钛硅涂层对钴铬合金耐腐蚀性能的影响[J]. 上海口腔医学, 2011, 20(6): 595-597.
[11] Meyer RD, Meyer J, Taloumis LJ. Intraoral galvanic corrosion: literature review and case report[J]. J Prosthet Dent, 1993, 69(2): 141-143.
[12] 战德松, 吕娇, 赵奇, 等. 磁性附着体中铁素体不锈钢引起小鼠成纤维细胞L929的死亡模式[J]. 金属学报, 2006, 42(8): 892-896.
[13] 孟贺, 韩东, 战德松. 5种牙科合金对小鼠成纤维细胞毒性及凋亡相关基因表达的影响[J]. 中华口腔医学杂志, 2009, 44(8): 497-501.