纳米银颗粒联合氢氧化钙对饥饿期粪肠球菌生物膜的作用

上海口腔医学 ›› 2016, Vol. 25 ›› Issue (1): 11-15.

纳米银颗粒联合氢氧化钙对饥饿期粪肠球菌生物膜的作用

张富华，李矛，韦智君，赵兵

合肥市口腔医院牙体牙髓病二科，安徽合肥 230001

收稿日期:2015-04-09 修回日期:2015-07-06 出版日期:2016-02-25 发布日期:2016-03-09
通讯作者: 张富华，Tel：0551-62631607，E-mail：zfh1024@163.com
作者简介:张富华（1979-），男，硕士，住院医师
基金资助:
安徽省年度重点科研项目（11070403040）

The effect of a combined nanoparticulate/calcium hydroxide medication on the biofilm of Enterococcus faecalis in starvation phase

ZHANG Fu-hua, LI Mao, WEI Zhi-jun, ZHAO Bing.

Department of Endodontics, Hefei Stomatological Hospital. Hefei 230001, Anhui Province, China

Received:2015-04-09 Revised:2015-07-06 Online:2016-02-25 Published:2016-03-09

摘要/Abstract

摘要： 目的： 将氢氧化钙和纳米银联合，探讨其对饥饿期粪肠球菌生物膜的抑制效果。方法： 使用256个牙根样本建立饥饿期粪肠球菌生物膜体外模型，使用平板菌落计数法和结晶紫染色法测定不同药物[Ca(OH)2+AgNP，Ca(OH)2 , AgNP]/不同作用时间（1、7 d）对饥饿期粪肠球菌生物膜的抑制效果。以灭菌水作为实验对照组。采用SPSS13.0软件包对数据进行统计学分析。结果： 在1 d和7 d时，Ca(OH)2+AgNP对粪肠球菌生物膜的抑制效果强于Ca(OH)2和AgNP；同时， AgNP对粪肠球菌生物膜的抑制效果显著强于Ca(OH)2。Ca(OH)2在7 d时对粪肠球菌生物膜的抑制效果强于1 d时；Ca(OH)2+AgNP和AgNP在7 d和1 d时对粪肠球菌生物膜的抑制效果无显著差异。结论： 纳米银颗粒联合氢氧化钙对饥饿期粪肠球菌生物膜具显著的抑制作用。

关键词: 氢氧化钙, 纳米银, 生物膜, 粪肠球菌

Abstract: PURPOSE: To evaluate the efficacy of Ca(OH)2 with a silver nanoparticle suspension to eliminate the biofilm of Enterococcus faecalis (E. faecalis) in starvation phase. METHODS： The biofilm models of E.faecalis in the starvation phase were built in vitro with 256 extracted human single-rooted teeth, and the methods of plate culture count and crystal violet biofilm assay were applied to determine the effect of the inhibition of different intracanal medicament (silver nanoparticle with calcium hydroxide, silver nanoparticle alone, calcium hydroxide alone ) to the biofilm of E. faecalis in starvation phase at 1 and 7 days. The negative control group was treated with sterile water only. Statistical analysis was carried out with SPSS 13.0 software package. RESULTS: The inhibitory effect of Ca（OH）2 + nanosilver on the biofilms of E. faecalis was found more significant than that of silver nanoparticle alone and calcium hydroxide alone at 1 and 7 days, and silver nanoparticle alone was more effective than calcium hydroxide alone. No difference in antimicrobial properties was observed between the two time points in the Ca(OH)2 + silver nanoparticle group and silver nanoparticle group, while higher antimicrobial efficacy was observed in the Ca(OH)2 group after 7 days than 1 day. CONCLUSIONS: Silver nanoparticle with calcium hydroxide has an obvious inhibitory effect on the biofilm of E.faecalis in the starvation phase.

Key words: Calcium hydroxide, Silver nanoparticle, Biofilm, Enterococcus faecalis

中图分类号:

R781.3

张富华，李矛，韦智君，赵兵. 纳米银颗粒联合氢氧化钙对饥饿期粪肠球菌生物膜的作用[J]. 上海口腔医学, 2016, 25(1): 11-15.

ZHANG Fu-hua, LI Mao, WEI Zhi-jun, ZHAO Bing.. The effect of a combined nanoparticulate/calcium hydroxide medication on the biofilm of Enterococcus faecalis in starvation phase[J]. Shanghai Journal of Stomatology, 2016, 25(1): 11-15.

参考文献

[1] Kakehashi S, Stanley HR, Fitzgerald RJ. The effects of surgical exposures of dental pulps in germ-free and conventional laboratory rats [J]. Oral Surg Oral Med Oral Pathol,1965, 20: 340-349.
[2] Evans M, Davies JK, Sundqvist G, et al. Mechanisms involved in the resistance of Enterococcus faecalis to calcium hydroxide [J].Int Endod J,2002,35(3):221-228.
[3] Sathorn C, Parashos P, Messer H. Australian endodontists’ perceptions of single and multiple visit root canal treatment [J].Int Endod J,2009, 42(9): 811-818.
[4] Turk BT, Sen BH, Ozturk T. In vitro antimicrobial activity of calcium hydroxide mixed with different vehicles against Enterococcus faecalis and Candida albicans[J].Oral Surg Oral Med Oral Pathol Oral Radiol Endod, 2009, 108(2): 297-301.
[5] Siqueira JF Jr, de Uzeda M. Disinfection by calcium hydroxide pastes of dentinal tubules infected with two obligate and one facultative anaerobic bacteria[J]. J Endod, 1996,22(12):674-676.
[6] Heling I, Steinberg D, Kenig S, et al. Efficacy of a sustained-release device containing chlorhexidine and Ca（OH）2 in preventing secondary infection of dentinal tubules[J].Int Endod J,1992,25(1): 20-24.
[7] Siqueira JF Jr, Lopes HP. Mechanisms of antimicrobial activity of calcium hydroxide: a critical review[J]. Int Endod J,1999,32(5): 361-369.
[8] Kishen A, Shi Z, Shrestha A, et al. An investigation on the antibacterial and antibiofilm efficacy of cationic nanoparticulates for root canal disinfection [J]. J Endod, 2008, 34(12):1515-1520.
[9] Moraes SR, Siqueira JF Jr, Colombo AP, et al. Comparison of the effectiveness of bacterial culture, 16S rDNA directed polymerase chain reaction, and checkerboard DNA-dNA hybridization for detection of Fusobacterium nucleatum in endodontic infections[J]. J Endod,2002, 28(2): 86-89.
[10] Haugland RA, Siefring SC, Wymer LJ, et al. Comparison of Enterococcus measurements in freshwater at two recreational beaches by quantitative polymerase chain reaction and membrane filter culture analysis [J]. Water Res, 2005, 39(4): 559-568.
[11] Basrani B, Tjaderhane L, Santos JM, et al. Efficacy of chlorhexidine- and calcium hydroxide-containing medicaments against Enterococcus faecalis in vitro[J]. Oral Surg Oral Med Oral Pathol Oral Radiol Endod, 2003, 96(5): 618-624.
[12] Gomes BP, Souza SF, Ferraz CC, et al. Effectiveness of 2% chlorhexidinegel and calcium hydroxide against Enterococcus faecalis in bovine root dentine in vitro[J].Int Endod J,2003,36(4): 267-275.
[13] Behnen MJ, West LA, Liewehr FR, et al. Antimicrobial activity of several calcium hydroxide preparations in root canal dentin [J]. J Endod, 2001, 27(12): 765-767.
[14] Lynne RE, Liewehr FR, West LA, et al. In vitro antimicrobial activity of various medication preparations on E. faecalis in root canal dentin [J].J Endod,2003, 29(3):187-190.
[15] Awawdeh L, Al-Beitawi M, Hammad M. Effectiveness of propolis and calcium hydroxide as a short-term intracanal medicament against Enterococcus faecalis: a laboratory study[J]. Aust Endod J, 2009, 35(2): 52-58.
[16] Safavi KE, Spangberg LS, Langeland K. Root canal dentinal tubule disinfection [J]. J Endod, 1990, 16(5): 207-210.
[17] Sj?gren U, Figdor D, Sp?ngberg L, et al. The antimicrobial effect of calcium hydroxide as a short-term intracanal dressing [J]. Int Endod J, 1991, 24(3): 119-125.
[18] Bhardwaj SB, Mehta M, Gauba K. Nanotechnology: role in dental biofilms [J]. Indian J Dent Res, 2009, 20(4): 511-513.
[19] Sondi I, Salopek-Sondi B. Silver nanoparticles as antimicrobial agent: a case study on E. coli as a model for Gram-negative bacteria [J]. J Colloid Interface Sci, 2004, 275(1): 177-182.
[20] Amin RM, Mohamed MB, Ramadan MA, et al. Rapid and sensitive microplate assay for screening the effect of silver and gold nanoparticles on bacteria [J].Nanomedicine (Lond),2009,4(6): 637-643.
[21] 庄沛林, 高燕, 凌均棨, 等. 纳米银对牙本质表面粪肠球菌生物膜的杀菌作用 [J].中华口腔医学研究杂志(电子版), 2011, 5(5): 463-469.
[22] Morones-Ramirez JR, Winkler JA, Spina CS, et al. Silver enhances antibiotic activity against gram-negative bacteria [J]. Sci Transl Med, 2013, 5(190): 190ra181.