本文選題：感染根管 + 牙本質(zhì)　； 參考：《南京大學(xué)》2014年碩士論文

【摘要】：第一部分不同類(lèi)型感染根管內(nèi)的細(xì)菌定植狀態(tài)觀(guān)察目的：通過(guò)掃描電鏡觀(guān)察不同類(lèi)型感染根管內(nèi)細(xì)菌定植狀況。材料與方法：臨床上收集新鮮拔除的25顆離體牙,分為3組：牙髓炎組(A組)、8例；根尖周炎組(B組)、10例；根管治療失敗組(C組)、7例。拔除后,用刀片去凈殘留的牙周膜和其余附著物,生理鹽水沖洗干凈,用2.5%磷酸戊二醛固定4h后,用低速切割機(jī)在釉牙骨質(zhì)界下截除牙冠,高速金剛砂車(chē)針?biāo)鋮s下將標(biāo)本沿牙(根)長(zhǎng)軸預(yù)備兩條互相平行的縱形深溝,溝深約2mm,用口外牙挺沿預(yù)備的凹槽輕撬將標(biāo)本一分為二。樣本經(jīng)4℃,0.1mol/L PBS溶液漂洗,乙醇梯度脫水(50%,60%,70%,80%,90%,100%梯度遞增,每次15min),二氧化碳臨界點(diǎn)干燥,離子濺射儀噴金后置掃描電鏡(日本日立S-2250N,電壓20kV)下觀(guān)察。每個(gè)根管分為根管冠1/3段(離根尖9mm),根管中1/3段(離根中6mm),根管尖1/3段(離根尖3mm),用解剖刀做定位,從低倍到高倍對(duì)根管全長(zhǎng)、根管壁、牙本質(zhì)小管進(jìn)行觀(guān)察；記錄觀(guān)察結(jié)果并照像。結(jié)果：A組：細(xì)菌分布主要限于根管冠1/3段牙本質(zhì)壁(75%),未進(jìn)入牙本質(zhì)小管內(nèi)。B組：細(xì)菌分布于根管全長(zhǎng)的牙本質(zhì)壁,可見(jiàn)細(xì)菌侵入牙本質(zhì)小管。C組：細(xì)菌分布主要集中于根管根尖1/3段牙本質(zhì)壁,大多以生物膜形式存在,多數(shù)牙本質(zhì)小管塌陷,可見(jiàn)細(xì)菌不同深度得定植在在牙本質(zhì)壁。細(xì)菌侵入牙本質(zhì)小管的深度比較：在根管中段,BC兩組沒(méi)有明顯差異(P0.05)；在根管尖段,C組細(xì)菌侵入牙本質(zhì)小管深度較B組深(P0.05)。結(jié)論：牙髓炎細(xì)菌感染主要局限于根冠部,細(xì)菌為侵入牙本質(zhì)小管；根尖周炎細(xì)菌感染可分布于根管全長(zhǎng),細(xì)菌進(jìn)入牙本質(zhì)小管內(nèi)；根管治療失敗的根管感染以根尖為主,以生物膜形式存在,細(xì)菌定植于牙本質(zhì)壁內(nèi)。第二部分不同類(lèi)型感染根管內(nèi)的4種厭氧菌的檢出分析目的：應(yīng)用16S rDNA-PCR技術(shù)檢測(cè)不同類(lèi)型感染根管內(nèi)4種厭氧菌定植狀況。材料與方法：臨床上收集新鮮拔除30顆離體牙,分為3組：牙髓炎組(A組)、10例；根尖周炎組(B組)、10例；根管治療失敗組(C組)、10例。拔除后,用刀片去凈殘留的牙周膜和其余附著物,生理鹽水沖洗干凈,用低速切割機(jī)在釉牙骨質(zhì)界下截除牙冠,然后用解剖刀做定位,在垂直于根管長(zhǎng)軸的方向上,每個(gè)根管分為根管冠中段(離根尖6mm),根管尖段(離根尖3mm),用無(wú)菌金剛砂片進(jìn)行截?cái)啵好總�(gè)根管截?cái)嗟谋砻媸褂眠^(guò)氧化氫和次氯酸鈉溶液消毒,接著使用5%硫代硫酸鈉溶擦洗；放入裝有l(wèi)ml的凍存管中,保存在液氮24h后,用高溫高壓滅菌的高密度白布包裹,錘子敲碎成碎塊,所有碎塊組織收集到裝有1mlPBS的1.5m1離心管中,-80度保存。提取樣本細(xì)菌基因組DNA,用PCR擴(kuò)增細(xì)菌16S rDNA基因片段的方法檢測(cè)細(xì)菌種類(lèi),計(jì)算檢出率。結(jié)果：通用引物檢測(cè)30病例,都能檢測(cè)到待檢細(xì)菌,檢出率達(dá)100%。在A(yíng)組,根管冠中段能檢測(cè)到牙髓卟啉單胞菌P. endodontalis (P.e)、具核梭桿菌F. nucleatum (F. n)和中間普氏菌P. intermedia (P. i)三種細(xì)菌,其中P.e的檢出率最高,而根管尖段只檢測(cè)到P.e的存在,而且檢出率比較低(10%),明顯低于根管冠中段(P0.05)。在B組,在檢出P. e、F. n、P. i的同時(shí),我們還檢測(cè)出糞腸球菌E. faecalis (E. f),在根管冠中段的檢出率分別是30%、20%、70%和10%和在根管尖段的檢出率分別是40%、40%、80%和10%,都無(wú)明顯統(tǒng)計(jì)學(xué)差異(P0.05),其中都是P.i的檢出率最高。在C組,E.f在根管尖段細(xì)菌的檢出率(60%)比在根管冠中段細(xì)菌的檢出率(10%)高,兩者之間具有明顯的統(tǒng)計(jì)學(xué)意義(P0.05),其他三種細(xì)菌的檢出率都比較低。結(jié)論：根管感染是多重細(xì)菌感染；其中這4種細(xì)菌是感染根管的優(yōu)勢(shì)菌；牙髓炎的根管冠中段以牙髓卟啉單胞菌為主,根管尖段細(xì)菌檢出率很低；根尖周炎的感染根管內(nèi)細(xì)菌在根管冠中段和根管尖段的4種細(xì)菌檢出率無(wú)明顯差別,都是以中間普氏菌為主,根管治療失敗的根管冠中段的4種細(xì)菌檢出率均很低,根管尖段的糞腸球菌檢出率最高。
[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.
【學(xué)位授予單位】：南京大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類(lèi)號(hào)】：R780.2

【共引文獻(xiàn)】

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