DNaseⅠ与Nd：YAP激光联合应用对根管内粪肠球菌生物膜清除作用的影响

doi:10.11958/20220653

天津医药 ›› 2023, Vol. 51 ›› Issue (1): 58-61.doi: 10.11958/20220653

DNaseⅠ与Nd：YAP激光联合应用对根管内粪肠球菌生物膜清除作用的影响

叶静¹(), 刘青², 温特¹, 杜瑞¹, 王园¹, 马贲¹, 尼娜¹^,^∆()

1 天津市天津医院口腔科（邮编300211）
2 天津医科大学口腔医院牙体牙髓科

收稿日期:2022-04-27 修回日期:2022-05-24 出版日期:2023-01-15 发布日期:2023-01-17
通讯作者: 尼娜 E-mail:812299853@qq.com;nina_tj@163.com
作者简介:叶静（1989），女，主治医师，主要从事根管内细菌生物膜方面研究。E-mail：812299853@qq.com

Effects of DNase I combined with Nd:YAP laser on the removal of enterococcus faecalis biofilm in root canals

YE Jing¹(), LIU Qing², WEN Te¹, DU Rui¹, WANG Yuan¹, MA Ben¹, NI Na¹^,^∆()

1 Department of Stomatology, Tianjin Hospital, Tianjin 300211, China
2 Department of Endodontics, School of Stomatology, Tianjin Medical University

Received:2022-04-27 Revised:2022-05-24 Published:2023-01-15 Online:2023-01-17
Contact: NI Na E-mail:812299853@qq.com;nina_tj@163.com

叶静, 刘青, 温特, 杜瑞, 王园, 马贲, 尼娜. DNaseⅠ与Nd：YAP激光联合应用对根管内粪肠球菌生物膜清除作用的影响[J]. 天津医药, 2023, 51(1): 58-61.

YE Jing, LIU Qing, WEN Te, DU Rui, WANG Yuan, MA Ben, NI Na. Effects of DNase I combined with Nd:YAP laser on the removal of enterococcus faecalis biofilm in root canals[J]. Tianjin Medical Journal, 2023, 51(1): 58-61.

摘要/Abstract

摘要：

目的研究脱氧核糖核酸酶Ⅰ（DNaseⅠ）与Nd：YAP激光联合应用对根管内粪肠球菌（E.f）生物膜的清除效果。方法建立牙本质片表面E.f感染模型，应用扫描电子显微镜（SEM）观察感染模型建立情况。将25个培养48 h的E.f生物膜样本采用随机数字表法分为5组，每组5个。A组：0.5%NaOCl;B组：0.5%NaOCl+Nd：YAP激光;C组：DNaseⅠ+0.5%NaOCl;D组：DNaseⅠ+0.5%NaOCl+Nd：YAP激光;E组：生理盐水。激光共聚焦显微镜（CLSM）观察牙本质表面E.f黏附情况，并计数绿色荧光量即为活细菌量。结果 SEM观察可见牙本质表面牙本质小管口开放，E.f黏附聚集在牙本质表面，并见到牙本质小管深层内细菌黏附。CLSM观察可见A、B、C组CLSM图像中绿色荧光的活细菌散在分布于牙本质表面，D组可见较少量的绿色荧光，E组绿色荧光的活细菌覆盖全部样本表面。D组中菌量明显小于A组和E组，C组菌量明显小于E组（P<0.01）。结论相比于单独使用0.5% NaOCl，DNaseⅠ联合Nd：YAP激光与0.5%NaOCl冲洗能更有效地处理E.f生物膜。

关键词: 粪肠球菌, 生物膜, 脱氧核糖核酸酶Ⅰ, Nd：YAP激光, 根管冲洗

Abstract:

Objective To study the effect of DNaseⅠ combined with Nd: YAP laser on the removal of enterococcus faecalis (E.f) biofilm in root canals. Methods The E.f infection model on dentin surface was established and observed by scanning electron microscope (SEM). E.f biofilm samples cultured for 48 h were divided into 5 groups by random number table method, with 5 samples in each group. The group A was treated with 0.5%NaOCl, the group B was treated with 0.5%NaOCl+Nd: YAP laser, the group C was treated with DNaseⅠ+ 0.5%NaOCl, the group D was treated with DNaseⅠ+ 0.5%NaOCl+Nd: YAP laser and the group E was treated with 0.9% normal saline. The adhesion of E.f on dentin surface was observed by laser confocal microscope (CLSM), and the amount of green fluorescence was counted, which was the amount of viable bacteria. Results SEM observation showed the dentin tubule opening on the dentin surface. E.f adhesion was observed on the dentin surface, and bacterial adhesion was observed in the deep layer of dentin tubule. CLSM observation showed that living bacteria with green fluorescence were scattered on the dentin surface in the group A, the group B and the group C. Less green fluorescence was observed in the group D. Green fluorescent live bacteria covered all the sample surface in the group E. The amount of bacteria in the group D was significantly less than that in the groups A and the groups E, and the amount of bacteria in the group C was significantly less than that in the group E (P<0.01). Conclusion Compared with 0.5% NaOCl alone, DNaseⅠ combined with Nd: YAP laser and 0.5% NaOCl can treat E.f biofilms more effectively.

Key words: Enterococcus faecalis, biofilms, deoxyribonuclease Ⅰ, Nd: YAP laser, root canal irrigation

中图分类号:

R781.3

图/表 3

图1 牙本质片表面粪肠球菌单菌种感染模型的建立

Fig.1 Establishment of a single E.f infection model on dentin discs

图2 牙本质表面感染模型的SEM图像

Fig.2 SEM images of dentin surface infection model

图3 不同方式处理后牙本质表面粪肠球菌E.f黏附的CLSM图像（SYTO-9/PI染色，×200）

Fig.3 CLSM images of E.f adhesion on dentin surface after different ways of processing (SYTO-9/PI staining, ×200)

参考文献 18

[1]	LI Q, ZHANG Q, ZOU X, et al. Evaluation of four final irrigation protocols for cleaning root canal walls[J]. Int J Oral Sci, 2020, 12(1):29. doi:10.1038/s41368-020-00091-4.
[2]	BOUTSIOUKIS C, ARIAS-MOLIZ M T. Present status and future directions - irrigants and irrigation methods[J]. Int Endod J, 2022, Suppl 3:588-612. doi:10.1111/iej.13739.
[3]	LIU T, HUANG Z, JU Y, et al. Bactericidal efficacy of three parameters of Nd:YAP laser irradiation against Enterococcus faecalis compared with NaOCl irrigation[J]. Lasers Med Sci, 2019, 34(2):359-366. doi:10.1007/s10103-018-2603-9.
[4]	CHRISTO J E, ZILM P S, SULLIVAN T, et al. Efficacy of low concentrations of sodium hypochlorite and low-powered Er,Cr:YSGG laser activated irrigation against an Enterococcus faecalis biofilm[J]. Int Endod J, 2016, 49(3):279-286. doi:10.1111/iej.12447.
[5]	SIQUEIRA J F Jr, RÔÇAS I N. Present status and future directions:Microbiology of endodontic infections[J]. Int Endod J, 2022, 55 (Suppl 3):512-530. doi:10.1111/iej.13677.
[6]	WHITCHURCH C B, TOLKER-NIELSEN T, RAGAS P C, et al. Extracellular DNA required for bacterial biofilm formation[J]. Science, 2002, 295(5559):1487. doi:10.1126/science.295.5559.1487.
[7]	OKSHEVSKY M, REGINA V R, MEYER R L. Extracellular DNA as a target for biofilm control[J]. Curr Opin Biotechnol, 2015, 33:73-80. doi:10.1016/j.copbio.2014.12.002.
[8]	CHENG F, ZHU Y. The efficacy of different instruments combined with Nd:YAP in endodontic retreatment[J]. Ann Transl Med, 2021, 9(14):1141. doi:10.21037/atm-21-2373.
[9]	SANGALLI J, JÚNIOR E, BUENO C, et al. Antimicrobial activity of Psidium cattleianum associated with calcium hydroxide against Enterococcus faecalis and Candida albicans:An in vitro study[J]. Clin Oral Investig, 2018, 22(6):2273-2279. doi:10.1007/s00784-017-2326-5.
[10]	RAMARAJ S, KIM M A, ROSA V, et al. Combined effect of melittin and DNase on enterococcus faecalis biofilms and its susceptibility to sodium hypochlorite[J]. Materials(Basel), 2020, 13(17):3740. doi:10.3390/ma13173740.
[11]	FORNAINI C, BRULAT-BOUCHARD N, MEDIONI E, et al. Nd:YAP laser in the treatment of dentinal hypersensitivity:An ex vivo study[J]. J Photochem Photobiol B, 2020, 203:111740. doi:10.1016/j.jphotobiol.2019.111740.
[12]	NAGAHASHI T, YAHATA Y, HANDA K, et al. Er:YAG laser-induced cavitation can activate irrigation for the removal of intraradicular biofilm[J]. Sci Rep, 2022, 12(1):4897. doi:10.1038/s41598-022-08963-x.
[13]	GOLOB B S, OLIVI G, VRABEC M, et al. Efficacy of photon-induced photoacoustic streaming in the reduction of enterococcus faecalis within the root canal:different settings and different sodium hypochlorite concentrations[J]. J Endod, 2017, 43(10):1730-1735. doi:10.1016/j.joen.2017.05.019.
[14]	SRIKANTH R, BANU S F, SOWNDARYA J, et al. Biosurfactant synergized with marine bacterial DNase disrupts polymicrobial biofilms[J]. Folia Microbiol (Praha), 2021, 66(5):831-842. doi:10.1007/s12223-021-00876-y.
[15]	PORTENIER I, WALTIMO T, ØRSTAVIK D, et al. The susceptibility of starved,stationary phase,and growing cells of Enterococcus faecalis to endodontic medicaments[J]. J Endod, 2005, 31(5):380-386. doi:10.1097/01.don.0000145421.84121.c8.
[16]	PANLILIO H, RICE C V. The role of extracellular DNA in the formation,architecture,stability,and treatment of bacterial biofilms[J]. Biotechnol Bioeng, 2021, 118(6):2129-2141. doi:10.1002/bit.27760.
[17]	YU M K, KIM M A, ROSA V, et al. Role of extracellular DNA in Enterococcus faecalis biofilm formation and its susceptibility to sodium hypochlorite[J]. J Appl Oral Sci, 2019, 27:e20180699. doi:10.1590/1678-7757-2018-0699.
[18]	PETRIDIS X, BUSANELLO F H, SO M, et al. Chemical efficacy of several NaOCl concentrations on biofilms of different architecture:new insights on NaOCl working mechanisms[J]. Int Endod J, 2019, 52(12):1773-1788. doi:10.1111/iej.13198.

DNaseⅠ与Nd：YAP激光联合应用对根管内粪肠球菌生物膜清除作用的影响

Effects of DNase I combined with Nd:YAP laser on the removal of enterococcus faecalis biofilm in root canals

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