本文選題：光動力療法 + 單態(tài)氧�。� 參考：《天津醫(yī)科大學(xué)》2014年碩士論文

【摘要】：目的： 根管消毒是根管治療中的關(guān)鍵步驟,其目的是徹底清除根管內(nèi)的感染源、預(yù)防再感染,目前根管消毒還未達(dá)到非常理想的程度,因此有必要尋找一種更有效、徹底的根管消毒方法。光動力療法(photodynamic therapy, PDT)是光、光敏劑和氧共同參與,可產(chǎn)生強氧化性的單態(tài)氧殺傷致病微生物的一種新療法,它具有抗菌作用強、抗菌譜廣、作用迅速、安全性好且不會產(chǎn)生耐藥性等優(yōu)點。EDTA(乙二胺四乙酸二鈉)作為根管沖洗劑具有滲透性強、能去除根管內(nèi)玷污層等優(yōu)點。本研究結(jié)合光動力療法與EDTA的優(yōu)點將二者聯(lián)合應(yīng)用,首先對根管內(nèi)感染的主要致病菌懸液進(jìn)行作用,篩選出二者聯(lián)合時光動力療法的最佳效應(yīng)參數(shù),然后在體外建立根管感染模型,探究二者聯(lián)合進(jìn)行根管消毒的效果,期望找到一種更加安全、有效的根管消毒方法。方法： 1.將糞腸球菌標(biāo)準(zhǔn)株(E. faecalis ATCC29212)復(fù)蘇厭氧培養(yǎng)48h,調(diào)整濃度制備成1×109CFU/mL的菌懸液。 2.菌懸液經(jīng)17%EDTA處理后,用波長532nm半導(dǎo)體激光器分別照射30s、60s、90s、120s與不同濃度光敏劑血卟啉單甲醚(hematoporphyrin monomethylether, HMME)20μg/ml、40μg/ml、60μg/ml進(jìn)行作用,選擇出EDTA預(yù)處理后光動力療法的最佳參數(shù)。 3.收集因正畸拔除的離體前磨牙75顆,采用冠向下法進(jìn)行根管預(yù)備,將糞腸球菌在根管內(nèi)連續(xù)培養(yǎng)21天建立糞腸球菌感染模型。 4.將感染模型隨機(jī)均分為5組,A組：PDT處理組；B組：EDTA處理組；C組：PDT聯(lián)合EDTA處理組；D組：NaClO陽性對照組；E組：生理鹽水陰性對照組。各組處理前、處理后分別用紙尖在根管內(nèi)取樣,進(jìn)行平板菌落計數(shù),統(tǒng)計分析,評價PDT聯(lián)合EDTA進(jìn)行根管消毒的即刻效果。 5.進(jìn)行回復(fù)實驗,在處理后的根管內(nèi)注滿無菌液體培養(yǎng)基,培養(yǎng)7天后用紙尖取樣,統(tǒng)計檢出細(xì)菌的根管個數(shù),統(tǒng)計分析,評價PDT聯(lián)合EDTA進(jìn)行根管消毒的遠(yuǎn)期效果。 6.掃描電子顯微鏡觀察處理后的根管內(nèi)壁形態(tài),進(jìn)一步驗證PDT聯(lián)EDTA進(jìn)行根管消毒的效果。結(jié)果： 1.在本實驗條件下,糞腸球菌菌懸液經(jīng)17%EDTA預(yù)處理后光動力療法的最佳效應(yīng)參數(shù)為：光敏劑HMME濃度40μg/ml；波長532nm、功率密度0.14W/cm2半導(dǎo)體激光器連續(xù)照射90s。 2.PDT聯(lián)合17%EDTA對感染根管內(nèi)糞腸球菌殺傷結(jié)果顯示,與生理鹽水陰性對照組相比,PDT處理組、EDTA處理組、PDT聯(lián)合EDTA處理組、NaC1O陽性對照組根管內(nèi)糞腸球菌數(shù)目均顯著減少(P0.05)；單獨PDT組殺菌率為89.77%,與NaClO陽性對照組相比無顯著差異(P0.05)；PDT聯(lián)合EDTA組殺菌率達(dá)到100%,明顯高于單獨PDT處理組、單獨EDTA處理組、NaClO陽性對照組和生理鹽水陰性對照組(P0.05)。 3.回復(fù)實驗結(jié)果顯示,生理鹽水陰性對照組與單獨EDTA處理組中全部根管都檢出了細(xì)菌,有菌根管比例達(dá)100%；單獨PDT處理組和NaClO陽性對照組檢出細(xì)菌根管個數(shù)分別為12個和13個,有菌根管比例分別為80.00%和86.67%；PDT聯(lián)合EDTA組檢出細(xì)菌根管數(shù)僅為4個,有菌根管比例僅為26.67%,明顯少于單獨PDT處理組、單獨EDTA處理組、NaClO陽性對照組和生理鹽水陰性對照組(P0.05)。 4.掃描電子顯微鏡圖像顯示：PDT聯(lián)合EDTA組根管內(nèi)壁無玷污層及細(xì)菌附著,全部牙本質(zhì)小管口呈開放狀態(tài)；牙本質(zhì)小管通暢,無玷污栓阻塞,內(nèi)壁光滑無細(xì)菌附著。 結(jié)論： 1.光動力療法聯(lián)合EDTA對體外游離的糞腸球菌有很好的殺滅作用,糞腸球菌菌懸液經(jīng)17%EDTA預(yù)處理后光動力療法的最佳效應(yīng)參數(shù)為：光敏劑HMME濃度40μg/ml；波長532nm、功率密度0.14W/cm2半導(dǎo)體激光器連續(xù)照射90s。 2.光動力療法聯(lián)合EDTA進(jìn)行根管消毒,不但能去除根管內(nèi)玷污層,而且還能有效殺滅根管內(nèi)糞腸球菌,并且有較好的遠(yuǎn)期效果,是一種高效的根管消毒方法,有望為提高臨床根管消毒效率提供一條新途徑。
[Abstract]:Objective:
Root canal disinfection is a key step in root canal therapy. The aim is to thoroughly remove the source of infection in the root canal and prevent reinfection. The root canal disinfection has not reached a very ideal level. Therefore, it is necessary to find a more effective and thorough root canal disinfection method. Photodynamic therapy (PDT) is a common reference to light, photosensitizer and oxygen. It can produce strong oxidizing mono oxygen to kill pathogenic microorganisms. It has the advantages of strong antibacterial effect, broad antibacterial spectrum, rapid action, good safety and no resistance to resistance..EDTA (ethylenediamine tetra acetic acid two) has the advantages of strong permeability and can remove the stain layer in root canal. The advantages of dynamic therapy and EDTA are combined with the two people. First, the main pathogenic bacteria suspension in the root canal infection is used, and the best effect parameters of the two combined time dynamic therapy are screened out, then the root canal infection model is established in vitro, and the effect of the two joint root canal elimination is explored, hoping to find a safer and more effective method. Method of root canal disinfection.
1. the E. faecalis ATCC29212 strain of Enterococcus faecium was resuscitation in anaerobic culture 48h and adjusted to 1 * 109CFU/mL suspension.
After the 2. bacteria suspension was treated by 17%EDTA, the wavelength 532nm semiconductor laser was used to irradiate 30s, 60s, 90s, 120s and the 20 micron (hematoporphyrin monomethylether, HMME) monomethyl ether (hematoporphyrin monomethylether, HMME) with different concentration of photosensitizer (hematoporphyrin monomethylether, HMME), and 60 micron g/ml. The best parameters of the photodynamic therapy after pretreatment were selected.
3. a total of 75 anterior molars extracted from orthodontic were collected, and the root canal preparation was carried out by the crown method. The Enterococcus faecalis was cultured in the root canal for 21 days to establish a model of Enterococcus faecalis infection.
4. group of infection models were randomly divided into 5 groups, group A: PDT treatment group; group B: EDTA treatment group; C group: PDT combined with EDTA treatment group; D group: NaClO positive control group; E group: normal saline negative control group. Before treatment, each group was treated with paper tip in root canal to carry out flat plate count, statistical analysis and evaluation PDT combined EDTA. The immediate effect of sterilizing the root canal.
5. the recovery experiment was carried out. The root canal was filled with the culture medium of sterile liquid in the root canal after treatment. After 7 days of culture, the sample was sampled with paper tip. The number of root canal of bacteria was detected statistically. The long-term effect of PDT combined with EDTA for root canal disinfection was evaluated.
6. scanning electron microscope (SEM) was used to observe the morphology of the root canal, and further verify the effect of root canal disinfection with PDT EDTA.
1. under the conditions of this experiment, the best effect parameters of photodynamic therapy for Enterococcus faecalis suspension after 17%EDTA preconditioning are: the concentration of photosensitizer HMME is 40 g/ml, the wavelength is 532nm, and the power density 0.14W/cm2 semiconductor laser irradiate 90s. continuously.
The killing results of 2.PDT combined with 17%EDTA in infected root canal Enterococcus showed that compared with the negative control group, the number of Enterococcus faecalis in the NaC1O positive control group decreased significantly (P0.05) in the PDT treatment group, the EDTA treatment group and the PDT combined EDTA treatment group, and the single PDT group was 89.77%, and there was no significant difference compared with the NaClO positive control group. In group EDTA, the bactericidal rate of PDT combined with EDTA was 100%, which was significantly higher than that of the single PDT treatment group, EDTA treatment group, NaClO positive control group and normal saline negative control group (P0.05).
3. the results of the 3. recovery test showed that all the root canals in the normal saline negative control group and the single EDTA treatment group were detected bacteria, and the proportion of the mycorrhizal tube was 100%. The number of bacterial root canal in the single PDT treatment group and the NaClO positive control group were 12 and 13 respectively, and the proportion of the mycorrhizal tube was 80% and 86.67%; the PDT combined with the EDTA group detected the bacteria. The root canal number was only 4, and the proportion of mycorrhizal tube was only 26.67%, obviously less than the single PDT treatment group, the single EDTA treatment group, the NaClO positive control group and the normal saline negative control group (P0.05).
4. scanning electron microscope images showed that the inner wall of PDT combined with EDTA group had no tarnished layer and bacteria attachment, all dentinal tubules were open, the dentinal tubules were unobstructed, no tarnishing embolus was blocked, and the inner wall was smooth and no bacteria attached.
Conclusion:
1. photodynamic therapy combined with EDTA has a good killing effect on the free Enterococcus faecalis in vitro. The best effect parameters of photodynamic therapy after 17%EDTA pretreatment are as follows: the HMME concentration of photosensitizer is 40 g/ml, the wavelength is 532nm, and the power density 0.14W/cm2 semiconductor laser is continuously irradiated with 90s.
2. photodynamic therapy combined with EDTA for root canal disinfection can not only remove the smear layer in the root canal, but also effectively kill the Enterococcus faecalis in the root canal, and has a good long-term effect. It is an efficient method of root canal disinfection. It is expected to provide a new way to improve the efficiency of clinical root canal disinfection.
【學(xué)位授予單位】：天津醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2014
【分類號】：R781.05

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