二氧化钛纳米粒子联合LED光照射对口腔变异链球菌的抑制作用及其机制  

Inhibitory effect of Streptococcus mutans by titanium dioxide nanoparticles combined with LED light irradiation and its mechanism

作　　者：闫琳琳 朱琳 赵远航 宋嘉卓 邹馨颖 刘新 赵红 张志民[1] 张红[1] YAN Linlin;ZHU Lin;ZHAO Yuanhang;SONG Jiazhuo;ZOU Xinying;LIU Xin;ZHAO Hong;ZHANG Zhimin;ZHANG Hong(Department of Endodontics,Stomatology Hospital,Jilin University,Changchun 130021,China;Jilin Provincial International Cooperation Base of Oral Biomedicine,Changchun 130021,China;State Key Laboratory of Supramolecular Structure and Materials,Jilin University,Changchun 130021,China)

机构地区：[1]吉林大学口腔医院牙体牙髓科,吉林长春130021 [2]吉林省口腔生物医学国际联合研究中心,吉林长春130021 [3]吉林大学超分子结构与材料国家重点实验室,吉林长春130012

出　　处：《吉林大学学报（医学版）》2022年第4期929-937,共9页Journal of Jilin University：Medicine Edition

基　　金：吉林省发改委2020年省预算内基本建设资金项目(2020C029-2);吉林省财政厅科技项目(JCSZ2020304-2)。

摘　　要：目的:探讨对二氧化钛纳米粒子(TiO_(2)NPs)进行锌(Zn)和氮(N)共掺杂时最适掺杂浓度及Zn和N共掺杂TiO_(2)NPs(Zn-N-TiO_(2) NPs)对口腔变异链球菌的抑制作用,阐明其作用机制。方法:采用钛酸丁酯、硝酸锌和氨水分别作为钛(Ti)源、Zn源和N源,利用溶胶-水热法合成TiO_(2) NPs和不同掺杂浓度的Zn-N-TiO_(2) NPs,通过X射线衍射(XRD)、拉曼光谱、扫描电子显微镜(SEM)和紫外-可见光吸收光谱分析(UV-vis)对纳米粒子进行表征,采用Cure Rite辐射仪对牙科发光二极管(LED)光固化灯进行表征。变异链球菌分为空白对照组、单独LED光照组、TiO_(2)NPs组、1%Zn-3%N-TiO_(2)NPs(Zn1)组、3%Zn-3%N-TiO_(2)NPs(Zn3)组、5%Zn-3%N-TiO_(2)NPs(Zn5)组和7%Zn-3%N-TiO_(2)NPs(Zn7)组。将各组纳米粒子与变异链球菌菌液混合(终浓度为2 g·L^(-1))摇匀。除空白对照组,其他各组采用牙科LED光分别照射1、3和5 min,空白对照组仅用培养基处理,采用平板菌落计数法记录各组平板菌落数。选取抗菌效果最佳组(Zn3组),利用1,3-二苯基异苯并呋喃(DPBF)探针检测活性氧(ROS)释放量,分为0 min组、1 min组、3 min组和5 min组,每组取2 g·L^(-1)的Zn3-DPBF溶液100μL分别采用牙科LED光照0、1、3和5 min,观察各组溶液在波长410 nm处吸收峰值,即代表ROS释放量。变异链球菌分为对照组、N-乙酰半胱氨酸(NAC)组、Zn3组和NAC+Zn3组,LED光照射5 min后避光培养24 h,采用平板菌落计数法计数各组平板菌落数。结果:成功合成Zn-N-TiO_(2)NPs,晶体结构为锐钛矿型,TiO_(2)NPs组、Zn1组、Zn3组、Zn5组和Zn7组纳米粒子平均晶粒尺寸分别为15.6、11.3、9.8、9.4和7.3 nm;SEM观察纳米粒子为分布均匀的球形颗粒;UV-vis显示Zn3在400~500 nm波长范围内吸光度(A)值明显升高。与空白对照组比较,LED光照射5 min时Zn3组菌落数最少(P<0.05)。与0 min组比较,1 min组、3 min组和5 min组Zn3-DPBF溶液在波长410 nm处吸收峰值依次下降,表明各Objective:To investigate the optimum doping concentrations of zinc(Zn)and nitrogen(N)in co-doping of titanium dioxide nanoparticles(TiO_(2)NPs)and the inhibitory effect of co-doping of Zn,N and TiO_(2)NPs(Zn-N-TiO_(2)NPs)on the Streptococcus mutans,and to clarify its mechanism.Methods:The TiO_(2)NPs and Zn-N-TiO_(2)NPs with different doping concentrations were synthesized by sol-hydrothermal method using butyl titanate,zinc nitrate and ammonia as titanium source,Zn source and N source,respectively.X-ray diffraction(XRD),Raman spectra,scanning electron microscope(SEM),UV-visible absorption spectroscope(UV-vis)were used to characterize the nanoparticles in various groups.Cure Rite radiometer was used to characterize the dental light emitting diode curing lamp.The Streptococcus mutans were divided into blank control group,LED light irradiation group,TiO_(2)NPs group,1%Zn-3%N-TiO_(2)NPs group(Zn1 group),3%Zn-3%N-TiO_(2)NPs group(Zn3 group),5%Zn-3%N-TiO_(2)NPs group(Zn5 group),and 7%Zn-3%N-TiO_(2)NPs group(Zn7 group).The nanoparticles in each group were mixed with Streptococcus mutans bacteria solution(the final concentration of nanoparticles was 2 g·L^(-1))and were shaken well.The nanoparticles groups and the LED light irradiation group were exposed to dental LED light for 1,3,and 5 min,the blank control group was treated with culture medium only,and the number of colonies in each group was recorded by plate colony counting method.The group with the best antibacterial effect(Zn3 group)was selected.The reactive oxygen species(ROS)release amount was detected using DPBF probe,and the experiment was divided into 0 min group,1 min group,3 min group and 5 min group.A total of 100μL Zn3-DPBF solution at the concentration of 2 g·L^(-1) in each group was selected for dental LED light irradiation for 0,1,3 and 5 min,respectively,and the changes in absorption peaks at 410 nm were observed,indicating the ROS release amount.In the experiment exploring the influence of N-acetyl cystethione(NAC)in the inhibitory effect of Zn3 co

关 键 词：二氧化钛 掺杂 变异链球菌 抗菌性能 

分 类 号：R781.1[医药卫生—口腔医学]