根管預(yù)備對(duì)感染根管形成根尖生物膜影響的體外研究
發(fā)布時(shí)間：2018-08-17 10:19
【摘要】：牙髓根尖周病是一類主要由細(xì)菌感染引起的、發(fā)生于牙髓及根尖周圍組織的炎癥性疾病，是口腔常見多發(fā)病，占口腔內(nèi)科門診的60%。根管治療術(shù)是保存患牙，治療牙髓根尖周病的首選方法。目前根管治療的成功率為68~85%。根管治療失敗主要表現(xiàn)為難治性根尖周炎和根管治療后疾病，而根尖生物膜被認(rèn)為是導(dǎo)致難治性根尖周炎和根管治療后疾病的主要原因。 目前較多的學(xué)者研究了根尖生物膜的結(jié)構(gòu)組成、來源，根尖生物膜來源于感染根管已達(dá)成共識(shí)，但對(duì)于感染根管形成根尖周炎及根尖生物膜機(jī)制的研究甚少，是否與拔髓、根管預(yù)備有關(guān)，尚無定論。根尖周標(biāo)本的采集方法主要依靠拔除患牙和根尖外科手術(shù)，但是能通過這兩種方法取材的病例有限，限制了對(duì)上述問題的研究。因此本實(shí)驗(yàn)擬建立根管-根尖周復(fù)合體體外模型，從體外研究根管預(yù)備對(duì)感染根管形成根尖生物膜的影響，以期指導(dǎo)臨床治療。 目的： 1.建立根管-根尖周復(fù)合體體外模型。 2.體外研究根管預(yù)備對(duì)感染根管形成根尖生物膜的影響。 方法： 1.將因正畸減數(shù)拔除的健康單根前磨牙密封于盛有LB固體培養(yǎng)基的無菌小瓶內(nèi)，使牙根的根尖1/3置于培養(yǎng)基中，制備成根管-根尖周復(fù)合體體外模型25個(gè)。建模后即刻隨機(jī)抽取5個(gè)模型通過PCR技術(shù)檢測(cè)根尖周有無細(xì)菌。將余下的20個(gè)模型隨機(jī)分為對(duì)照組（n=10）及實(shí)驗(yàn)組（n=10），實(shí)驗(yàn)組做開髓處理，對(duì)照組不做任何處理。自然放置于空氣中，在建模后21d檢測(cè)根管內(nèi)和根尖周有無細(xì)菌，及根尖周內(nèi)毒素（終點(diǎn)顯色法）。 2.按照上述方法制作根管-根尖周復(fù)合體體外模型（n=115）。于建模后即刻隨機(jī)抽取5個(gè)模型通過PCR技術(shù)檢測(cè)根尖周有無細(xì)菌。將余下的110個(gè)模型隨機(jī)分為對(duì)照組（n=25）和實(shí)驗(yàn)組（n=85），對(duì)照組不做處理，實(shí)驗(yàn)組離體牙開髓，模型暴露于空氣中21天。實(shí)驗(yàn)組隨機(jī)抽取5個(gè)模型以檢測(cè)根管內(nèi)有無細(xì)菌存在，確定已形成感染根管；對(duì)照組隨機(jī)抽取5個(gè)模型以檢測(cè)根尖周有無細(xì)菌，以確定模型的封閉效果。將實(shí)驗(yàn)組余下的80個(gè)模型分為4組。第1組不做處理，第2組拔髓，第3組按離體牙工作長度預(yù)備根管，第4組超出根尖孔預(yù)備根管。根管預(yù)備后，所有模型均放置于空氣中。根管預(yù)備后即刻、7d、14d、21d用PCR檢測(cè)根尖周有無細(xì)菌，若檢測(cè)到細(xì)菌則使用掃描電子顯微鏡觀察離體牙根尖外表面有無根尖生物膜形成；根管預(yù)備后即刻、7d、21d用終點(diǎn)顯色法檢測(cè)根尖周內(nèi)毒素含量。 結(jié)果： 1.在建模后21d，實(shí)驗(yàn)組根管內(nèi)檢測(cè)到細(xì)菌，對(duì)照組根管內(nèi)未檢測(cè)到細(xì)菌，而所有模型根尖區(qū)均未檢測(cè)到細(xì)菌；實(shí)驗(yàn)組根尖周內(nèi)毒素含量增加,與對(duì)照組比較，經(jīng)統(tǒng)計(jì)學(xué)分析差異有統(tǒng)計(jì)學(xué)意義（P0.01）。 2.根管預(yù)備后即刻、7d、14d，根尖周均未檢測(cè)到細(xì)菌；根管預(yù)備后21d，，第3組和第4組根尖周檢測(cè)到細(xì)菌，在離體牙根尖外表面通過掃描電鏡觀察到生物膜的存在。與第1組相比，預(yù)備后即刻，第2組根尖周內(nèi)毒素含量的差異無統(tǒng)計(jì)學(xué)意義（P0.05）；而第3組和第4組，根尖周內(nèi)毒素含量減少，第4組減少更顯著，經(jīng)統(tǒng)計(jì)學(xué)分析差異有統(tǒng)計(jì)學(xué)意義(P0.01)。預(yù)備后7天、21天，與第1組相比，第2組根尖周內(nèi)毒素含量的差異無統(tǒng)計(jì)學(xué)意義；第3組和第4組根尖周內(nèi)毒素含量增加顯著，第4組增加更顯著，經(jīng)統(tǒng)計(jì)學(xué)分析差異有統(tǒng)計(jì)學(xué)意義（P0.01）。 結(jié)論： 1.成功建立根管-根尖周復(fù)合體體外模型。不干擾感染根管時(shí)，感染根管內(nèi)的細(xì)菌不易到達(dá)根尖周，而感染根管內(nèi)的細(xì)菌首先是通過分泌的內(nèi)毒素等致病因子到達(dá)根尖周引起根尖周炎。 2.全長預(yù)備或者超出根尖孔預(yù)備根管比只拔髓更容易導(dǎo)致感染根管內(nèi)的細(xì)菌擴(kuò)散至根尖周，且根尖周的內(nèi)毒素含量增加更顯著。
[Abstract]:Periodontal disease is a kind of inflammatory disease mainly caused by bacterial infection. It occurs in the pulp and periapical tissues. It is a common oral disease, accounting for 60% of the outpatient department of oral medicine. Refractory periapical periodontitis and disease after root canal therapy are the main causes of refractory periapical periodontitis and disease after root canal therapy.
At present, many scholars have studied the structure and composition of the apical biofilm. The origin of the apical biofilm is from the infected root canal. There is a consensus that the apical biofilm originates from the infected root canal. However, little research has been done on the mechanism of periapical inflammation and biofilm formation in the infected root canal. In order to guide the clinical treatment, a root canal-periapical complex model was established to study the effect of root canal preparation on the formation of apical biofilm in vitro.
Objective:
1. to establish an in vitro model of root canal periapical complex.
2. the effect of root canal preparation on root canal biofilm formation was studied in vitro.
Method:
1. The healthy single premolar extracted by orthodontic subtraction was sealed in a sterile vial containing LB solid medium. One third of the root tip was placed in the culture medium. 25 in vitro models of the root canal-periapical complex were prepared. Five models were randomly selected to detect the presence or absence of bacteria in the periapical zone immediately after modeling. The machine was divided into control group (n = 10) and experimental group (n = 10). The experimental group was treated with pulpotomy, and the control group was not treated with any treatment.
2. In vitro model of root canal-periapical complex (n=115) was made according to the above method. Five models were randomly selected to detect bacteria around the apex by PCR immediately after modeling. The remaining 110 models were randomly divided into control group (n=25) and experimental group (n=85). The control group was not treated, the experimental group was pulp-opening, and the model was exposed to air 21. The experimental group was randomly divided into four groups: the first group was not treated, the second group was pulped, and the third group was treated according to the isolated teeth. After root canal preparation, all the models were placed in the air. The bacteria around the apex were detected by PCR immediately, 7 days, 14 days and 21 days after root canal preparation. If bacteria were detected, the apical biofilm formation was observed by scanning electron microscope. 7d and 21d were used to detect endotoxin content in periapical by terminal color test.
Result:
1. Bacteria were detected in the root canals of the experimental group and the control group 21 days after the model was established. Bacteria were not detected in the root canals of the control group, but not in all the apical regions of the model.
2. Bacteria were not detected in the periapical area immediately after root canal preparation, 7 days and 14 days, and bacteria were detected in the periapical area of the third and fourth groups 21 days after root canal preparation, and biofilm was observed on the external surface of the root apex in vitro by scanning electron microscope. There was no significant difference in endotoxin content between the third group and the fourth group, especially in the fourth group (P 0.01). The difference was statistically significant (P0.01).
Conclusion:
1. Successful establishment of an in vitro model of the root canal-periapical complex. Without interfering with the infected root canals, the bacteria in the infected root canals are not easy to reach the periapical periodontitis, but the bacteria in the infected root canals reach the periapical periodontitis by secreting endotoxins and other pathogenic factors.
2. The bacteria in the infected root canals were more likely to spread to the periapical zone and the endotoxin content in the periapical zone increased more significantly than that in the pulp extraction alone.
【學(xué)位授予單位】：重慶醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：R781.05
[Abstract]:Periodontal disease is a kind of inflammatory disease mainly caused by bacterial infection. It occurs in the pulp and periapical tissues. It is a common oral disease, accounting for 60% of the outpatient department of oral medicine. Refractory periapical periodontitis and disease after root canal therapy are the main causes of refractory periapical periodontitis and disease after root canal therapy.
At present, many scholars have studied the structure and composition of the apical biofilm. The origin of the apical biofilm is from the infected root canal. There is a consensus that the apical biofilm originates from the infected root canal. However, little research has been done on the mechanism of periapical inflammation and biofilm formation in the infected root canal. In order to guide the clinical treatment, a root canal-periapical complex model was established to study the effect of root canal preparation on the formation of apical biofilm in vitro.
Objective:
1. to establish an in vitro model of root canal periapical complex.
2. the effect of root canal preparation on root canal biofilm formation was studied in vitro.
Method:
1. The healthy single premolar extracted by orthodontic subtraction was sealed in a sterile vial containing LB solid medium. One third of the root tip was placed in the culture medium. 25 in vitro models of the root canal-periapical complex were prepared. Five models were randomly selected to detect the presence or absence of bacteria in the periapical zone immediately after modeling. The machine was divided into control group (n = 10) and experimental group (n = 10). The experimental group was treated with pulpotomy, and the control group was not treated with any treatment.
2. In vitro model of root canal-periapical complex (n=115) was made according to the above method. Five models were randomly selected to detect bacteria around the apex by PCR immediately after modeling. The remaining 110 models were randomly divided into control group (n=25) and experimental group (n=85). The control group was not treated, the experimental group was pulp-opening, and the model was exposed to air 21. The experimental group was randomly divided into four groups: the first group was not treated, the second group was pulped, and the third group was treated according to the isolated teeth. After root canal preparation, all the models were placed in the air. The bacteria around the apex were detected by PCR immediately, 7 days, 14 days and 21 days after root canal preparation. If bacteria were detected, the apical biofilm formation was observed by scanning electron microscope. 7d and 21d were used to detect endotoxin content in periapical by terminal color test.
Result:
1. Bacteria were detected in the root canals of the experimental group and the control group 21 days after the model was established. Bacteria were not detected in the root canals of the control group, but not in all the apical regions of the model.
2. Bacteria were not detected in the periapical area immediately after root canal preparation, 7 days and 14 days, and bacteria were detected in the periapical area of the third and fourth groups 21 days after root canal preparation, and biofilm was observed on the external surface of the root apex in vitro by scanning electron microscope. There was no significant difference in endotoxin content between the third group and the fourth group, especially in the fourth group (P 0.01). The difference was statistically significant (P0.01).
Conclusion:
1. Successful establishment of an in vitro model of the root canal-periapical complex. Without interfering with the infected root canals, the bacteria in the infected root canals are not easy to reach the periapical periodontitis, but the bacteria in the infected root canals reach the periapical periodontitis by secreting endotoxins and other pathogenic factors.
2. The bacteria in the infected root canals were more likely to spread to the periapical zone and the endotoxin content in the periapical zone increased more significantly than that in the pulp extraction alone.
【學(xué)位授予單位】：重慶醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：R781.05
【參考文獻(xiàn)】
相關(guān)期刊論文 前5條
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