[1]马莹莹,邹朝晖.光动力疗法对大鼠龋齿模型的防龋效果及机制[J].新乡医学院学报,2018,35(1):016-21.[doi:10.7683/xxyxyxb.2018.01.004]
　MA Ying-ying,ZOU Zhao-hui.Effect and mechanism of photodynamic therapy on dental caries in rats[J].Journal of Xinxiang Medical University,2018,35(1):016-21.[doi:10.7683/xxyxyxb.2018.01.004]
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光动力疗法对大鼠龋齿模型的防龋效果及机制
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《新乡医学院学报》[ISSN:1004-7239/CN:41-1186/R]

卷:
35
期数:
2018年1
页码:
016-21
栏目:
基础研究
出版日期:
2018-01-05

文章信息/Info

Title:
Effect and mechanism of photodynamic therapy on dental caries in rats
作者:
马莹莹1; 邹朝晖2
（1.郑州市口腔医院牙体牙髓科，河南　郑州　450000；2.天津医科大学口腔医学院牙体牙髓科，天津　300070）
Author(s):
MA Ying-ying1; ZOU Zhao-hui2
（1.Department of Dentistry，Zhengzhou Stomatological Hospital，Zhengzhou 450000，Henan Province，China；2.Department of Dentistry，School of Stomatology，Tianjin Medical University，Tianjin 300070，China）
关键词:
大鼠; 龋齿; 血卟啉单甲醚; 光动力疗法; Keyes计分
Keywords:
rat; dental caries; hematoporyrin monomethyl ether; photodynamic therapy; Keyes score
分类号:
R781.1
DOI:
10.7683/xxyxyxb.2018.01.004
文献标志码:
A
摘要:
目的　观察光动力疗法（PDT）对大鼠龋齿模型的防龋效果，并探讨其防龋机制。方法　将以变异链球菌为致龋菌建立大鼠龋齿模型成功的40只21 d鼠龄、断奶的无特定病原体级Wistar雄性大鼠随机分为生理盐水组、血卟啉单甲醚（HMME）-PDT组、单纯激光组、单纯光敏剂组和氟化钠组，每组8只。生理盐水组大鼠用浸有150 μL生理盐水的小棉球反复涂搽磨牙各面5 min；HMME-PDT组大鼠用浸有150 μL 40 mg·L-1 HMME的小棉球放于磨牙各牙面上，避光孵育5 min后，用532 nm的半导体激光垂直照射磨牙各面，功率密度为140 mW·cm-2，照射时间为 90 s，能量密度为12.6 J·cm-2；单纯激光组大鼠不进行光敏剂HMME处理，仅用532 nm的半导体激光垂直照射磨牙各面，照射时间、功率密度、能量密度均同HMME-PDT组；单纯光敏剂组大鼠仅用浸有150 μL 40 mg·L-1 HMME的小棉球放于磨牙各牙面上，避光孵育5 min，不进行激光照射；氟化钠组大鼠用浸有150 μL 2 g·L-1氟化钠溶液的小棉球反复涂搽磨牙各面5 min；各组大鼠每周处理1次，连续4周，每次处理后及时采集大鼠口腔变异链球菌进行培养，平板菌落计数记录细菌抑制情况，连续4周后处死大鼠，记录龋齿Keyes计分，扫描电镜观察牙齿表面形态。结果　时间因素对大鼠口腔变异链球菌的生长无影响（F=0.11，P>0.05），处理因素对大鼠口腔变异链球菌的生长有影响（F=230.89，P< 0.05），且处理和时间因素存在交互效应（F=6.36，P<0.05）。第1、2、3、4周处理后，HMME-PDT组和氟化钠组大鼠牙齿变异链球菌菌落数均少于生理盐水组、单纯光敏剂组和单纯激光组（P<0.05）；第1、2、3周处理后，HMME-PDT组大鼠牙齿变异链球菌菌落数均少于氟化钠组（P<0.05）。各组大鼠平滑面龋齿均未检出Dx级龋损；HMME-PDT组和氟化钠组大鼠平滑面龋齿均未检出Dm级龋损。HMME-PDT组、氟化钠组大鼠平滑面龋齿E、Ds级龋损的Keyes计分均显著低于生理盐水组、单纯光敏剂组和单纯激光组（P<0.05），但HMME-PDT组与氟化钠组大鼠平滑面龋齿E、Ds级龋损的Keyes计分比较差异无统计学意义（P>0.05）。各组大鼠窝沟龋齿中，仅生理盐水组检出Dx级龋损。HMME-PDT组、氟化钠组大鼠窝沟龋齿E、Ds、Dm 级龋损Keyes计分均显著低于生理盐水组、单纯光敏剂组和单纯激光组（P<0.05），但HMME-PDT组与氟化钠组大鼠之间窝沟龋齿E、Ds、Dm 级龋损的Keyes计分比较差异无统计学意义（P>0.05）。扫描电镜显示，生理盐水组和单纯光敏剂组大鼠牙釉质表面粗糙，凸凹不平，有大量凹坑状脱矿区和划痕；单纯激光组大鼠牙釉质表面相对光滑，有少量圆形凹坑并可见凹坑内熔融状结构；氟化钠组大鼠牙釉质表面相对光滑，脱矿区较少，并可见到脱矿区的再矿化；HMME-PDT组大鼠牙釉质表面较平整，点状凹陷减少，深脱矿区少，且脱矿区域大多表浅。结论　HMME-PDT可通过减少致龋菌数量、改善牙齿表面结构等途径对大鼠致龋模型起到明显的预防作用。
Abstract:
Objective　To evaluate the effect of photodynamic therapy （PDT） on dental caries in rats，and to investigate the mechanism.Methods　Forty male Wistar rats （21 days old，specific pathogen-free，weaned） with dental caries induced by Streptococcus mutans were randomly divided into saline group，hematoporphyrin monomethyl ether （HMME）-PDT group，simple laser group，simple photosensitizer group and sodium fluoride group，with eight rats in each group.The molars of rats in the saline group were smear repeatedly for 5 minutes with cotton balls containing 150 μL normal saline.The molars of rats in HMME-PDT group were incubated for 5 minutes without light by placing the cotton balls containing 150 μL HMME （40 mg·L-1） on the tooth surface，then molars were irradiated vertically by 532 nm semiconductor laser，the power density was 140 mW·cm-2，the irradiation time was 90 s，and the energy density was 12.6 J·cm-2.The molars of rats in the simple laser group were irradiated vertically by 532 nm semiconductor laser；the irradiation time，power density and energy density were the same as those in HMME-PDT group；but the molars were not treated with HMME.The molars of rats in the simple photosensitizer group were smear repeatedly for 5 minutes with cotton balls containing 150 μL HMME （40 mg·L-1），but the molars were not irradiated by laser.The molars of rats in the sodium fluoride group were smear repeatedly for 5 minutes with cotton balls containing 150 μL fluorinse （2 g·L-1）.The rats in each group were treated once a week for 4 weeks.Streptococcus mutans were collected from oral cavity of rats after each treatment，and bacterial inhibition was recorded by plate count.The rats were killed after 4 weeks，and the Keyes caries score was recorded.The surface morphology of the teeth was observed by scanning electron microscope.Results　The time factor had no effect on the growth of oral Streptococcus mutans （F=0.11，P>0.05），the treatment factor had effect on the growth of oral Streptococcus mutans （F=230.89，P<0.05），and there was interaction effect between treatment factor and time factor （F=6.36，P<0.05）.The colony number of Streptococcus mutans in HMME-PDT group and sodium fluoride group was lower than that in saline group，photosensitizer group and simple laser group at the first，second，third and fourth week after treatment （P<0.05）.The colony number of Streptococcus mutans in HMME-PDT group was lower than that in sodium fluoride group at the first，second and third week after treatment （P<0.05）.The Dx caries lesion of smooth surface caries wasn′t found in each group.The Dm caries lesion of smooth surface caries wasn′t found in the HMME-PDT group and sodium fluoride group.The Keyes scores of E and Ds caries lesion of smooth surface caries in the HMME-PDT group and sodium fluoride group was lower than that in the saline group，simple photosensitizer group and simple laser group（P<0.05），but there was no significant difference in the Keyes scores of E and Ds caries lesion of smooth surface caries between the HMME-PDT group and sodium fluoride group（P>0.05）.The Dx caries lesion of pit and fissure caries was found in the saline group only.The Keyes score of E，Ds and Dm caries lesion of pit and fissure caries in the HMME-PDT group and sodium fluoride group was lower than that in the saline group，simple photosensitizer group and simple laser group（P<0.05），but there was no significant difference in the Keyes scores of E，Ds and Dm caries lesion of pit and fissure caries between the HMME-PDT group and sodium fluoride group（P>0.05）.The results of scanning electron microscopy showed that the enamel surface was rough，uneven and there was a large number of scratch in the saline group and simple photosensitizer group.Relatively the enamel surface in the simple laser group was smooth，and there was a small circular scallops and dents was found in the structure；in the sodium fluoride group，the enamel surface was relatively smooth，take off the mining area was less，and remineralization was found.In the HMME-PDT group，the enamel surface was smooth，pitting was reduced and take off the mining area was small.Conclusion　HMME-PDT had significant inhibitive efficacy on caries development in rats by reducing the number of cariogenic bacteria and improving the structure of the teeth.

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更新日期/Last Update: 2018-01-05