不同类型感染根管内的细菌学分析

发布时间：2018-06-28 14:23

本文选题：感染根管 + 牙本质　；参考：《南京大学》2014年硕士论文

【摘要】：第一部分不同类型感染根管内的细菌定植状态观察目的：通过扫描电镜观察不同类型感染根管内细菌定植状况。材料与方法：临床上收集新鲜拔除的25颗离体牙,分为3组：牙髓炎组(A组)、8例；根尖周炎组(B组)、10例；根管治疗失败组(C组)、7例。拔除后,用刀片去净残留的牙周膜和其余附着物,生理盐水冲洗干净,用2.5%磷酸戊二醛固定4h后,用低速切割机在釉牙骨质界下截除牙冠,高速金刚砂车针水冷却下将标本沿牙(根)长轴预备两条互相平行的纵形深沟,沟深约2mm,用口外牙挺沿预备的凹槽轻撬将标本一分为二。样本经4℃,0.1mol/L PBS溶液漂洗,乙醇梯度脱水(50%,60%,70%,80%,90%,100%梯度递增,每次15min),二氧化碳临界点干燥,离子溅射仪喷金后置扫描电镜(日本日立S-2250N,电压20kV)下观察。每个根管分为根管冠1/3段(离根尖9mm),根管中1/3段(离根中6mm),根管尖1/3段(离根尖3mm),用解剖刀做定位,从低倍到高倍对根管全长、根管壁、牙本质小管进行观察；记录观察结果并照像。结果：A组：细菌分布主要限于根管冠1/3段牙本质壁(75%),未进入牙本质小管内。B组：细菌分布于根管全长的牙本质壁,可见细菌侵入牙本质小管。C组：细菌分布主要集中于根管根尖1/3段牙本质壁,大多以生物膜形式存在,多数牙本质小管塌陷,可见细菌不同深度得定植在在牙本质壁。细菌侵入牙本质小管的深度比较：在根管中段,BC两组没有明显差异(P0.05)；在根管尖段,C组细菌侵入牙本质小管深度较B组深(P0.05)。结论：牙髓炎细菌感染主要局限于根冠部,细菌为侵入牙本质小管；根尖周炎细菌感染可分布于根管全长,细菌进入牙本质小管内；根管治疗失败的根管感染以根尖为主,以生物膜形式存在,细菌定植于牙本质壁内。第二部分不同类型感染根管内的4种厌氧菌的检出分析目的：应用16S rDNA-PCR技术检测不同类型感染根管内4种厌氧菌定植状况。材料与方法：临床上收集新鲜拔除30颗离体牙,分为3组：牙髓炎组(A组)、10例；根尖周炎组(B组)、10例；根管治疗失败组(C组)、10例。拔除后,用刀片去净残留的牙周膜和其余附着物,生理盐水冲洗干净,用低速切割机在釉牙骨质界下截除牙冠,然后用解剖刀做定位,在垂直于根管长轴的方向上,每个根管分为根管冠中段(离根尖6mm),根管尖段(离根尖3mm),用无菌金刚砂片进行截断：每个根管截断的表面使用过氧化氢和次氯酸钠溶液消毒,接着使用5%硫代硫酸钠溶擦洗；放入装有lml的冻存管中,保存在液氮24h后,用高温高压灭菌的高密度白布包裹,锤子敲碎成碎块,所有碎块组织收集到装有1mlPBS的1.5m1离心管中,-80度保存。提取样本细菌基因组DNA,用PCR扩增细菌16S rDNA基因片段的方法检测细菌种类,计算检出率。结果：通用引物检测30病例,都能检测到待检细菌,检出率达100%。在A组,根管冠中段能检测到牙髓卟啉单胞菌P. endodontalis (P.e)、具核梭杆菌F. nucleatum (F. n)和中间普氏菌P. intermedia (P. i)三种细菌,其中P.e的检出率最高,而根管尖段只检测到P.e的存在,而且检出率比较低(10%),明显低于根管冠中段(P0.05)。在B组,在检出P. e、F. n、P. i的同时,我们还检测出粪肠球菌E. faecalis (E. f),在根管冠中段的检出率分别是30%、20%、70%和10%和在根管尖段的检出率分别是40%、40%、80%和10%,都无明显统计学差异(P0.05),其中都是P.i的检出率最高。在C组,E.f在根管尖段细菌的检出率(60%)比在根管冠中段细菌的检出率(10%)高,两者之间具有明显的统计学意义(P0.05),其他三种细菌的检出率都比较低。结论：根管感染是多重细菌感染；其中这4种细菌是感染根管的优势菌；牙髓炎的根管冠中段以牙髓卟啉单胞菌为主,根管尖段细菌检出率很低；根尖周炎的感染根管内细菌在根管冠中段和根管尖段的4种细菌检出率无明显差别,都是以中间普氏菌为主,根管治疗失败的根管冠中段的4种细菌检出率均很低,根管尖段的粪肠球菌检出率最高。
[Abstract]:The first part of the bacterial colonization in the root canal of different types of infection: through scanning electron microscopy to observe the bacterial colonization in the root canal of different types of infection. Materials and methods: 25 fresh extracted teeth were collected and divided into 3 groups: the pulpitis group (group A), 8 cases, the periapical periodontitis group (group B), and the root canal therapy failure group. C group), 7 cases, after extraction, use the blade to clean the residual periodontal membrane and the rest of the attachment, rinse the physiological saline, and use the 2.5% phosphate glutaraldehyde to fix the 4h, use the low speed cutting machine to remove the crown under the glazed cementum boundary, and prepare the specimens along the tooth (root) long shaft to prepare two parallel longitudinal deep gully with the long axis of the tooth (root), and the depth of the ditch is about 2mm. The specimens were divided into two parts along the prepared grooves. The samples were rinsed by 4 degrees centigrade, 0.1mol/L PBS solution, gradient dehydration of ethanol (50%, 60%, 70%, 80%, 90%, 100% gradient, each time 15min), the critical point of carbon dioxide was dried, and the ion sputter apparatus was observed under the scanning electric mirror (Japanese Hitachi S-2250N, voltage 20kV). Each root canal was divided into root canal. 1/3 segment (from root tip 9mm), 1/3 segment of root canal (6mm from root), 1/3 segment of root canal (3mm), using anatomic knife to locate, observe root canal full length, root canal wall, dentinal tubule from low to high times, record observation results and image. Results: A group: the distribution of bacteria is mainly limited to the dentine wall (75%) of root canal crown 1/3 segment (75%), not into dentine. .B group in the canaliculus: bacteria are distributed in the dentinal wall of the root canal, and bacteria invade the dentinal tubule.C group: the distribution of bacteria is mainly concentrated on the 1/3 segment of root canal root, most of which exist in the form of biofilm, most of the dentinal tubules collapse, and the bacteria can be colonized at the dentine wall at different depths. Bacteria invade the dentinal tubule. Depth comparison: in the middle part of the root canal, there was no significant difference in the BC two groups (P0.05). In the root canal, group C bacteria invaded the dentinal tubules deeper than the B group (P0.05). Conclusion: the bacterial infection of the pulpitis is mainly confined to the root crown, the bacteria are intruded into the dentinal tubule, and the bacterial infection of the root tip is distributed in the root canal, and the bacteria enter the dentine. In the canaliculus, root canal infection was mainly root canal infection, with biofilm in the form of biofilm, bacteria colonized in dentin wall. Second anaerobes in second different types of infection root canal were detected and analyzed: using 16S rDNA-PCR technique to detect 4 anaerobic bacteria colonization conditions in different types of infected root canals. Materials and methods: 30 fresh teeth were extracted and divided into 3 groups: the pulpitis group (group A), 10 cases, the periapical periodontitis group (group B), 10 cases, the root canal therapy failure group (group C) and 10 cases. After the extraction, the residual periodontal membrane and other attachment were removed with the blade, the saline was washed and dried, and the crown was cut off under the glazed cementum boundary with the low speed cutting machine, and then the solution was used. Each root canal was divided into the middle part of the root canal (6mm) and the root canal (from the root tip 3mm). The root canal was truncated with the aseptic sands: the surface of each root canal was truncated by hydrogen peroxide and sodium hypochlorite solution, and then 5% sodium thiosulfate was used to scrub; then LML was put into the tube. In the storage tube, after stored in the liquid nitrogen 24h, the high density white cloth wrapped in high temperature and high pressure was wrapped, the hammer was broken into fragments, all the fragments were collected in the 1.5m1 centrifuge tube with 1mlPBS, and -80 degree was preserved. The bacterial genome DNA was extracted and the bacterial 16S rDNA gene fragment was amplified by PCR to detect the bacterial species and calculate the detection rate. The results were as follows: the results were as follows: Using primers to detect 30 cases, the detected bacteria were detected, the detection rate was 100%. in group A, the P. endodontalis (P.e), F. nucleatum (F. n) and P. intermedia (P.) were detected in the middle segment of the root canal, among which the detection rate was the highest and the root canal pointed only the presence of the bacteria. And the detection rate was low (10%), obviously lower than that of the middle root canal (P0.05). In group B, P. e, F. n, P. I were detected, and we detected the E. faecalis (E. f) of Enterococcus faecalis, and the detection rates in the middle segment of the root canal were 30%, 20%, 70% and 10%, and the detection rates were 40%, 40%, 80% and 10% in the root canal, respectively. .05), among them, the detection rate of P.i was the highest. In group C, the detection rate of E.f in root canal tip bacteria (60%) was higher than that of bacteria in the root canal crown (10%), and there was significant statistical significance (P0.05), and the detection rates of other three kinds of bacteria were low. The dominant bacteria in the root canal; the middle segment of the root canal of the pulpitis was dominated by porphyromoninomonas pulp, and the detection rate of the root canal tip bacteria was very low; there were no significant differences in the detection rates of 4 bacteria in the root canal and root canal tip of the root canal. All were 4 kinds of root canal in the root canal. The detection rate of bacteria was very low, and the detection rate of Enterococcus faecalis was the highest in the apical segment.
【学位授予单位】：南京大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R780.2

【共引文献】