利用激光共聚焦扫描显微镜观察根管外细菌的分布和活性
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国家自然科学基金青年基金(81300868);江苏省高校自然科学研究项目(13KJB320008);江苏高校优势学科建设工程资助项目(2014-37)


Observation of extraradicular bacterial localization and viability by confocal laser scanning microscopy
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    摘要:

    目的:利用激光共聚焦扫描显微镜观察慢性根尖周炎患牙根尖区根管外细菌的分布范围和活性。方法:临床采集10例需拔除的伴有慢性根尖周炎的单根牙 (实验组) 和5例因正畸需要拔除的健康单根牙 (对照组)。所有样本首先经LIVE/DEAD■ BacLightTM死活菌染色试剂盒染色,各样本包埋后由根尖向冠方切取5张1 mm厚的切片。每张切片上随机选择3个视野,用激光共聚焦扫描显微镜观察每张切片上根管外细菌的分布情况和细菌的活性。结果:对照组中,牙骨质有较弱的荧光信号,通过设定观察条件,以对照组切片上无荧光信号为基线进行观察。实验组10例样本中,8例样本的根尖区切片的牙根外表面可观察到由显示绿色荧光信号的活菌和显示红色荧光信号的死菌共同构成的根管外生物膜。根管外细菌主要分布于距根尖2~3 mm范围内的根面牙骨质上。在68%的被观察视野中,活菌的绿色荧光的强度强于死菌的红色荧光。结论:根管外生物膜由活菌和死菌共同构成,分布于部分慢性根尖周炎患牙的根尖区牙骨质外表面上。

    Abstract:

    Objective:To verify the existence of extraradicular biofilm of teeth with radiographically visible chronic periapical lesion and to observe the distribution and viability of extraradicular bacteria using confocal laser scanning microscopy(CLSM). Methods:Ten single-rooted teeth with periapical radiolucent areas were collected. Five single-rooted teeth extracted for orthodontic reasons were selected as controls. All of the roots were embedded in methylmethacrylate after staining with LIVE/DEAD■ BacLightTM Bacterial Viability Kits. Then,five evenly distributed transverse sections(1 mm thick)were cut from the apex to the corona. Finally,the sections were examined by CLSM. Results:There were weak auto-fluorescence signals in the control group. After adjusting the viewing conditions to obtain a baseline,at which no fluorescence was detectable in the control group specimens,8 of the 10 disease group teeth showed positive fluorescence signals on the external root surfaces. The distribution of extraradicular bacteria was extensive within a 2~3 mm range from the root tip to the corona. On 68% of optical views,the intensity of green fluorescence signals was stronger than that of red fluorescence signals. Conclusion:There were live and dead extraradicular bacteria on the external root surfaces of teeth with chronic periapical periodontitis.

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唐志娟,王 娟.利用激光共聚焦扫描显微镜观察根管外细菌的分布和活性[J].南京医科大学学报(自然科学版),2015,(2):249-253

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  • 收稿日期:2014-04-23
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  • 在线发布日期: 2015-02-13
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