TBO-PDT对根管内粪肠球菌抑制作用的实验研究
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摘要
目的:体外观察不同光敏剂浓度和不同激光能量密度的TBO-PDT抑制离体牙根管内粪肠球菌的效果,确定最佳光敏剂参数和激光参数,并在此基础上探讨TBO-PDT在抑制离体牙根管内粪肠球菌上的作用优势,为PDT应用于根管消毒的临床实践提供理论与实验依据。方法:对离体牙进行根管预备,接种粪肠球菌,厌氧培养21d后建立粪肠球菌感染根管模型。将其随机分组:A组为阴性对照组,以生理盐水冲洗根管;B组为阳性对照组,以5.25%NaClO冲洗根管;其余组别根据光敏剂浓度的不同,分为C1组80μg/mL、D1组90μg/mL、E1组100μg/mL、F1组110μg/mL、G1组120μg/mL,避光孵育5min后用波长为630nm功率密度为90mW/cm2半导体激光在根管内上下提拉螺旋照射90s;根据光照时间的不同,分为C2组30s、D2组60s、E2组90s、F2组120s、G2组150s,光敏剂浓度为100μg/mL避光孵育5min后,用波长为630nm功率密度为90mW半导体激光在根管内上下提拉螺旋照射各实验组牙标本,实验处理前后即刻取样,厌氧培养24h,计数平板菌落,明确实验处理前后根管内粪肠球菌的数量变化。结果:(1)不同光敏剂浓度下TBO-PDT对根管内粪肠球菌抑菌效果的影响。平板菌落计数结果显示:不同光敏剂浓度的各组抑菌率随光敏剂浓度的增加而增高。与阴性对照组相比,C1组、D1组、E1组、F1组和G1组的抑菌率显著提高,而F1组和G1组两组间抑菌率没有明显差异。(2)不同光照时间对TBO-PDT抑制实验根管内以生物膜形式存在的粪肠球菌的抑菌效果。平板菌落计数结果显示:不同光照时间的各组抑菌率随光照时间延长而增高。与阴性对照组相比,其中C2组、D2组、E2组、F2组和G2组的抑菌率明显升高,而F2组和G2组两组间抑菌率没有明显差异。结论:TBO-PDT对以生物膜形式存在的粪肠球菌有明显的抑制作用,实验证实TBO-PDT最佳光敏剂浓度为110μg/mL,最佳光照时间为120s。
Objectives:The purpose of this study was to assess the antibacterial effect of TBO-photodynamic therapy on Enterococcus faecalis which infected extracted root canals and to determine the optimum photosensitizer concentration and light dose parameters.On that basis,it was designed to study the advantage of TBO-PDT on the antibacterial effects.This study provides theoretical and experimental evidence for PDT applied to clinical practice root canal disinfection.Methods:We made root canal preparation for experimental teeth.After preparation,the specimens were infected with Enterococcus faecalis for 21 dto establish a model of Enterococcus faecalis root canal infection.The specimens were randomly divided into twelve groups.Group A was irrigated with physiological saline.Group B was irrigated with 5.25%NaClO.For other groups,they were divided into five groups according to different concentrations of photosensitizer,as Group C1 80μg/ml、Group D1 90μg/mL、Group E1100μg/mL、Group F1 110μg/ml、Group G1 120μg/mL.The specimens were incubated for 5minutes in the dark,and then exposured to light at 630 nm 90mW for 90 sin a spiral pattern.According to the irradiation time,the other specimens were divided into five groups,as Group C2 30 s,Group D2 60 s,Group E2 90 s,Group F2 120 s,and Group G2 150 s.The specimens were incubated for 5minutes in the dark with a 100μg/mL concentration of photosensitizer,and then exposured to light at630 nm 90mW for certain time in a spiral pattern.Take Samples immediately before and after the experimental treatment.Anaerobic culture them for 24 hand count the colonies in the plates to clear the changes in the number of Enterococcus faecalis in experimental root canals after treatment.Results:(1)The antibacterial effects of TBO-PDT in different photosensitizer concentration groups.Plate colony count results showed that:Different photosensitizer concentration groups antibacterial rates were increased when the concentration of the photosensitizer was increased.Compared with the negative control group,the antibacterial rates of Group C1,D1,E1,F1 and G1were significantly increased.However,the antibacterial rates between Group F1 and G1 were not significantly different.(2)The antibacterial effects of TBO-PDT in different illumination time groups.Plate colony count results showed that:Different photosensitizer concentration groups antibacterial rates were increased with the illumination time extension.Compared with the negative control group,the antibacterial rates of Group C2,D2,E2,F2 and G2were significantly increased.However,the antibacterial rates between Group F2 and G2were not significantly different.Conclusion:TBO-PDT significantly inhibited Enterococcus faecalis which exist in the form of biofilm.Our study confirmed the best photosensitizer concentration of TBO-PDT is 110μg/ml,and the optimal illumination time is120 s.
Objectives:The purpose of this study was to assess the antibacterial effect of TBO-photodynamic therapy on Enterococcus faecalis which infected extracted root canals and to determine the optimum photosensitizer concentration and light dose parameters.On that basis,it was designed to study the advantage of TBO-PDT on the antibacterial effects.This study provides theoretical and experimental evidence for PDT applied to clinical practice root canal disinfection.Methods:We made root canal preparation for experimental teeth.After preparation,the specimens were infected with Enterococcus faecalis for 21 dto establish a model of Enterococcus faecalis root canal infection.The specimens were randomly divided into twelve groups.Group A was irrigated with physiological saline.Group B was irrigated with 5.25%NaClO.For other groups,they were divided into five groups according to different concentrations of photosensitizer,as Group C1 80μg/ml、Group D1 90μg/mL、Group E1100μg/mL、Group F1 110μg/ml、Group G1 120μg/mL.The specimens were incubated for 5minutes in the dark,and then exposured to light at 630 nm 90mW for 90 sin a spiral pattern.According to the irradiation time,the other specimens were divided into five groups,as Group C2 30 s,Group D2 60 s,Group E2 90 s,Group F2 120 s,and Group G2 150 s.The specimens were incubated for 5minutes in the dark with a 100μg/mL concentration of photosensitizer,and then exposured to light at630 nm 90mW for certain time in a spiral pattern.Take Samples immediately before and after the experimental treatment.Anaerobic culture them for 24 hand count the colonies in the plates to clear the changes in the number of Enterococcus faecalis in experimental root canals after treatment.Results:(1)The antibacterial effects of TBO-PDT in different photosensitizer concentration groups.Plate colony count results showed that:Different photosensitizer concentration groups antibacterial rates were increased when the concentration of the photosensitizer was increased.Compared with the negative control group,the antibacterial rates of Group C1,D1,E1,F1 and G1were significantly increased.However,the antibacterial rates between Group F1 and G1 were not significantly different.(2)The antibacterial effects of TBO-PDT in different illumination time groups.Plate colony count results showed that:Different photosensitizer concentration groups antibacterial rates were increased with the illumination time extension.Compared with the negative control group,the antibacterial rates of Group C2,D2,E2,F2 and G2were significantly increased.However,the antibacterial rates between Group F2 and G2were not significantly different.Conclusion:TBO-PDT significantly inhibited Enterococcus faecalis which exist in the form of biofilm.Our study confirmed the best photosensitizer concentration of TBO-PDT is 110μg/ml,and the optimal illumination time is120 s.
引文
[1]DOMENICO RICUCCI,HOHN RUSSO,MICHAEL RUTBERG,et al.A prospective cohort study of endodontic treatment of 1369root canals:results after 5years[J].Oral surgery,Oral Medicine,Oral Pathology,Oral Radiol and Endodontology,2011,112(6):825-842.
[2]GOMES B P,PINHEIRO E T,SOUSA E L,et al.Enterococcus faecalis in dental root canals detected by culture and by polymerase chain reaction analysis[J].Oral Surgery,Oral Medicine,Oral Pathology,Oral Radiology,and Endodontology,2006,102(2):247-253.
[3]DISTEL J W,HATTON J F,GILLESPIE M J.Biofilm Formation in Medicated Root Canals[J].Journal of Endodontic,2002,28(10):689-693.
[4]SOUKOS N S,CHEN P S Y,MORRIS J R,et a1.Photodynamic therapy for endodontic disinfection[J].Journal of Endodontic,2006,32(10):979-984.
[5]FOSCHI F,FONTANA C R,RUGGIERO K,et a1.Photodynamic inactivation of enterococcus faecalis in dental root canals in vitro[J].Lasers Surg Med,2007,39(10):782-787.
[6]KATIE FISHER,CAROL PHILLIPS.The ecology,epidemiology and virulence of Enterococcus facealis[J].Microbiology,2009,155(6):1749-1757.
[7]SOUZA L C,BRITO P R,OLIVEIRA J C,et a1.Photodynamic therapy with two diferent photosensitizers as a supplement to procedures in promo-ting intraeanal reduction of Enterococcus facealis[J].Jendod,2010,36(2):292-296.
[8]KRANZ S,GUELLMAR A,VOLPEL A,et al.Photodynamic suppression of enterococcus faecalis using the photosensitizer m THpC[J].Lasers in Surgery and Medicine,2011(43):241-248.
[9]SOUKOSN S,CHEN P S Y,MORRIS J T,et al.Photodynamic therapy for endodontic disinfection[J].Journal of Endodontics,2006,32(10):979-984.
[10]ALEJANDRO RIOS,JIANING HE,GERALD N,et al.Evaluation of photodynamic therapy using a light-emitting diode lamp against enterococcus faecalis in Extracted Human Teeth[J].Journal of Endodontics,2011,37(6):856-859.
[11]ZOLFAGHARI P S,SAMANTHA PACKER.In vivo killing of staphylococcus aureus using a light-activated antimicrobial agent[J].BMC Microbiology,2009,9(27):1-8.
[2]GOMES B P,PINHEIRO E T,SOUSA E L,et al.Enterococcus faecalis in dental root canals detected by culture and by polymerase chain reaction analysis[J].Oral Surgery,Oral Medicine,Oral Pathology,Oral Radiology,and Endodontology,2006,102(2):247-253.
[3]DISTEL J W,HATTON J F,GILLESPIE M J.Biofilm Formation in Medicated Root Canals[J].Journal of Endodontic,2002,28(10):689-693.
[4]SOUKOS N S,CHEN P S Y,MORRIS J R,et a1.Photodynamic therapy for endodontic disinfection[J].Journal of Endodontic,2006,32(10):979-984.
[5]FOSCHI F,FONTANA C R,RUGGIERO K,et a1.Photodynamic inactivation of enterococcus faecalis in dental root canals in vitro[J].Lasers Surg Med,2007,39(10):782-787.
[6]KATIE FISHER,CAROL PHILLIPS.The ecology,epidemiology and virulence of Enterococcus facealis[J].Microbiology,2009,155(6):1749-1757.
[7]SOUZA L C,BRITO P R,OLIVEIRA J C,et a1.Photodynamic therapy with two diferent photosensitizers as a supplement to procedures in promo-ting intraeanal reduction of Enterococcus facealis[J].Jendod,2010,36(2):292-296.
[8]KRANZ S,GUELLMAR A,VOLPEL A,et al.Photodynamic suppression of enterococcus faecalis using the photosensitizer m THpC[J].Lasers in Surgery and Medicine,2011(43):241-248.
[9]SOUKOSN S,CHEN P S Y,MORRIS J T,et al.Photodynamic therapy for endodontic disinfection[J].Journal of Endodontics,2006,32(10):979-984.
[10]ALEJANDRO RIOS,JIANING HE,GERALD N,et al.Evaluation of photodynamic therapy using a light-emitting diode lamp against enterococcus faecalis in Extracted Human Teeth[J].Journal of Endodontics,2011,37(6):856-859.
[11]ZOLFAGHARI P S,SAMANTHA PACKER.In vivo killing of staphylococcus aureus using a light-activated antimicrobial agent[J].BMC Microbiology,2009,9(27):1-8.