上海口腔医学

上海口腔医学 ›› 2014, Vol. 23 ›› Issue (1): 30-34.

• 基础研究 • 上一篇    下一篇

Ca(OH)2对牙髓卟啉单胞菌脂多糖细胞毒性的影响

郭佳杰1, 仇丽鸿1, 3, 于雅琼1, 徐莉雅1, 范昀倩1, 钟鸣2   

  1. 1.中国医科大学附属口腔医院 牙体牙髓病科, 辽宁 沈阳 110002;
    2.中国医科大学附属口腔医院 中心实验室, 辽宁 沈阳 110002;
    3.辽宁省口腔医学研究所, 辽宁 沈阳 110002
  • 收稿日期:2013-08-30 修回日期:2013-10-29 出版日期:2014-02-20 发布日期:2014-10-21
  • 通讯作者: 仇丽鸿,Tel：024-22895932, Fax: 024-22892645, E-mail: drqlh@yahoo.com
  • 作者简介:郭佳杰(1987-),男,硕士研究生,E-mail: jiajieguo@hotmail.com
  • 基金资助:
    辽宁省科学技术计划项目(2011225020)

Effect of Ca(OH)2 on the cytotoxicity of lipopolysaccharide extracted from Porphyromonas endodontalis in vitro

GUO Jia-jie1, QIU Li-hong1, 3, YU Ya-qiong1, XU Li-ya1, FAN Yun-qian1, ZHONG Ming2   

  1. 1.Department of Endodontics, College of Stomatology, China Medical University. Shenyang 110002, Liaoning Province, China;
    2.Central Laboratory, College of Stomatology, China Medical University. Shenyang 110002, Liaoning Province, China;
    3.Liaoning Institute of Dental Research. Shenyang 110002, Liaoning Province, China
  • Received:2013-08-30 Revised:2013-10-29 Online:2014-02-20 Published:2014-10-21
  • Supported by:
    Science and Technology Projects of Liaoning Province (2011225020)

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摘要： 目的：检测Ca(OH)2溶液在体外对牙髓卟啉单胞菌(Porphyromonas endodontalis,P.e)脂多糖(Lipopolysaccharide,LPS)的降解作用,并观察P.e LPS经Ca(OH)2 溶液作用后对小鼠成骨细胞MC3T3-E1增殖的影响,以完善 Ca(OH)2在根管消毒中的抑菌机制。方法：不同浓度Ca(OH)2溶液作用MC3T3-E1细胞5 d,甲基噻唑基四唑(MTT)法检测Ca(OH)2对MC3T3-E1细胞增殖的影响;P.e LPS与Ca(OH)2溶液体外分别作用30 min及60 min,显色基质法鲎试验检测P.e LPS的活性;P.e LPS与Ca(OH)2体外作用6 h,再作用MC3T3-E1细胞1、3和5 d,MTT法检测对MC3T3-E1细胞增殖的影响;采用SPSS 13.0软件包对数据进行统计学分析。结果：15%浓度以下的Ca(OH)2溶液显著促进MC3T3-E1细胞的增殖;10%和15% Ca(OH)2溶液体外作用P.e LPS 30 min和60 min,显著降低P.e LPS的活性;P.e LPS经15% Ca(OH)2溶液体外作用6 h后,不再抑制MC3T3-E1细胞的增殖。结论：Ca(OH)2溶液可以有效降解P.e LPS,并显著减弱P.e LPS对成骨细胞增殖的影响。

关键词: 氢氧化钙, 牙髓卟啉单胞菌, 脂多糖, 成骨细胞

Abstract: PURPOSE: To detect the degradation of Ca(OH)2 on lipopolysaccharide (LPS) extracted from Porphyromonas endodontalis (P.e) in vitro and estimate the influence of P.e LPS pretreated with Ca(OH)2 on the proliferation of MC3T3-E1 cells. METHODS: The effect of Ca(OH)2 on MC3T3-E1 cell proliferation was detected by methyl thiazolyl tetrazolium (MTT) assay. Then P.e LPS was treated with Ca(OH)2 for 30 mins or 60 mins at 37℃ in vitro and the activity of P.e LPS was evaluated by Chromogenic End-point Tachypleus Amebocyte Lysate (CE TAL) test. Finally, MC3T3-E1 cells were exposed to P.e LPS pretreated with 15% Ca(OH)2 for 1, 3 and 5 d, and the cell proliferation was measured using the MTT assay comparing with the P.e LPS control group. SPSS 13.0 software package was used for statistical analysis. RESULTS: Compared with the negative control, exposing cells to 5%, 10% and 15% Ca(OH)2 had greatly promoted MC3T3-E1 cell proliferation. P.e LPS treated with 10% and 15% Ca(OH)2 both presented the best results by CE TAL and significant difference compared with P.e LPS control group. When 10 μg/mL P.e LPS was pretreated with 15% Ca(OH)2, no inhibition of MC3T3-E1 cell proliferation was noted. CONCLUSIONS: Ca(OH)2 detoxifies P.e LPS in vitro, mitigates the impact of P.e LPS on MC3T3-E1 cell proliferation.

Key words: Calcium hydroxide, Lipopolysaccharide, Osteoblasts

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