Microflora profiling of infected root canal before and after treatment using culture-independent methods

This study aimed to profile the microflora in infected root canals before and after root canal treatment using culture-independent methods. Six infected root canals in single-rooted teeth with periapical lesions from five subjects were included. Quantification of total bacteria was performed by real-time PCR with primers targeting 16S rRNA genes. PCR products with universal 16S rRNA gene primers were cloned and partially sequenced, and bacterial identification at the species level was performed by comparative analysis with the GenBank database. The concentration of extracted DNA before treatment was higher than that after root canal treatment, although the difference was not statistically significant. Sequence analysis revealed that oral bacteria such as Fusobacterium, Streptococcus, Olsenella, and Pseudoramibacter detected in cases before root canal treatment disappeared after treatment. These results suggest that the root canal microflora are distinct before and after root canal treatment, and that treatment changes the microflora in both quantity and quality.     

两种根管冲洗液对狗牙根管内细菌抗菌作用的实验研究

目的观察2%洗必泰溶液和3%双氧水2种根管冲洗液对狗牙根管内粪肠球菌、溶血链球菌、微小消化链球菌、中间普氏菌及具核酸杆菌的影响,为临床根管治疗提供参考。方法选择成年健康杂种狗3只,共有24个实验牙,48个实验牙根。于狗牙根管内接种粪肠球菌、溶血链球菌、微小消化链球菌、中间普氏菌及具核酸杆菌。对狗牙完成根管治疗。于治疗后12个月拍摄根尖X线片,并记录牙齿和根尖周组织的临床表现。将患牙随机分为3组,每组7颗患牙,去除根管内充填物并进行根管预备,实验一组用2%洗必泰溶液冲洗根管,实验二组用3%双氧水冲洗根管,对照组用0.9%NaC l溶液冲洗根管。分别在根管预备前及根管预备冲洗后对根管内细菌取样,培养,鉴定并记录细菌菌落数量,测定根管内细菌变化情况。结果根管预备冲洗后3组根管内的细菌量均较根管预备前显著下降（P〈0.05）。2%洗必泰溶液和3%双氧水2种根管冲洗液的杀菌效果明显好于0.9%NaC l溶液（P〈0.05）,2%洗必泰溶液明显好于3%双氧水（P〈0.05）。结论2%洗必泰溶液是有效的根管冲洗药物,可明显减少狗牙根管内粪肠球菌、溶血链球菌、微小消化链球菌、中间普氏菌及具核酸杆菌的数量,但不能完全清除根管内的细菌。     

Microbiological study and scanning electron microscopic analysis of root canal wall dentin following pumped Diodium Nd:YAG laser irradiation

The capability of Nd:YAG laser in sterilizing root canals and the alterations of dentinal walls induced by laser treatment were investigated. Thirty root canals were infected by P. aeruginosa ATCC 27853 and thirty canals by A. naeslundii CH-12. Within each infection, 4 groups were selected on the basis of the treatment. Among them, 2 test groups (TGs) were treated by Nd:YAG laser at 15 Hz for 15 s, using 2 different settings: 1 Watt/70 Joule and 1.5 Watt/100 Joule, respectively (n = 10 each). The other 2 groups, used as controls (CGs), were: untreated (positive control, n = 5) and sterilized by 5.25% NaClO group (negative control, n = 5). Observations under scanning electron microscope (SEM) and quantitative bacterial counts were performed. These analyses were performed once per group after infections and treatments. Laser treatments significantly reduced the number of both bacteria. SEM investigation showed melting and crystallization of canal dentin over 1.5 W/100 J. Laser irradiation has a bactericidal effect but it does not completely sterilize the root canal as NaClO 5.25% solution does if the goal of treatment is also to avoid alterations of dentinal walls.     

奥硝唑用于感染根管消毒的细菌学研究

目的　观察和探讨新型抗厌氧菌药物奥硝唑消毒感染根管的抗菌效果。方法　选择感染根管患牙98颗,常规根管预备后,随机分为试验组(奥硝唑组)和对照组(甲醛甲酚组)各4 9颗。分别在根管预备前、预备后及封药后进行根管内细菌标本采样、培养、分离与鉴定,测定根管内细菌变化及检出情况,并观察封药后临床症状和体征的变化。结果　2组封药后,根管内细菌数量及检出率均较封药前明显降低( P<0 .0 5 ) ,根管内细菌数量及检出率比较,差异无显著性( P>0 .0 5 ) ,奥硝唑的近期临床疗效优于甲醛甲酚。结论　奥硝唑是一种较安全、有效的根管消毒药物     

Microbiota in the apical root canal system of tooth with apical periodontitis

Background

Apical periodontitis (AP) is essentially an inflammatory disease of microbial etiology primarily caused by infection of the pulp and root canal system. Variation of the bacterial communities caused by AP, as well as their changes responding to dental therapy, are of utmost importance to understand the pathogensis of the apical periodontitis and establishing effective antimicrobial therapeutic strategies. This study aims to uncover the composition and diversity of microbiota associated to the root apex to identify the relevant bacteria highly involved in AP, with the consideration of root apex samples from the infected teeth (with/without root canal treatment), healthy teeth as well as the healthy oral.

Methods

Four groups of specimens are considered, the apical part of root from diseased teeth with and without root canal treatment, and wisdom teeth extracted to avoid being impacted (tooth healthy control), as well as an additional healthy oral control from biofilm of the buccal mucosa. DNA was extracted from these specimens and the microbiome was examined through focusing on the V3-V4 hypervariable region of the 16S rRNA gene using sequencing on Illumina MiSeq platform. Composition and diversity of the bacterial community were tested for individual samples, and between-group comparisons were done through differential analysis to identify the significant changes.

Results

We observed reduced community richness and diversity in microbiota samples from diseased teeth compared to healthy controls. Through differential analysis between AP teeth and healthy teeth, we identified 49 OTUs significantly down-regulated as well as 40 up-regulated OTUs for AP.

Conclusion

This study provides a global view of the microbial community of the AP associated cohorts, and revealed that AP involved not only bacteria accumulated with a high abundance, but also those significantly reduced ones due to microbial infection.

     

Er,Cr:YSGG激光照射对根管封闭效果影响的实验研究

探讨常规根管预备后使用不同能量参数Er,C r:YSGG激光进行根管内照射对根管封闭效果的影响,为此激光在根管治疗领域的临床应用提供重要的实验依据。选择新拔除的单根管牙80颗,分成4组,每组20颗牙。常规根管预备后,分别使用1 W、2 W、3 W Er,C r:YSGG激光于有水条件下进行根管内照射,另选一组不做任何处理,作为阴性对照。扫描电子显微镜下观察根管内壁形态学改变后,常规根管充填,检测根管微渗漏情况。采用单因素方差分析及q检验进行统计学处理。结果可见:(1)形态学观察:与对照组相比,激光照射均可有效去除根管内壁的玷污层及碎屑;牙本质小管口清晰可见;未见熔融及炭化等热损伤改变;其中3 W激光处理后根管内壁形态最为理想。(2)微渗漏检测:与对照组相比,各激光组根管微渗漏深度均显著减少(P<0.05);但各激光组间比较无显著性差异(P>0.05)。综上,使用Er,C r:YSGG激光进行根管内照射可达到较为理想的根管清洁目的,显著改善根管充填后的封闭效果,因此有望在临床上应用提高根管治疗的成功率。     

Distinction of Prevotella intermedia and Prevotella nigrescens from endodontic bacteremia through their fatty acid contents

Prevotella nigrescens has recently been recognized as a new species distinct from Prevotella intermedia. The distinction is based largely on DNA-DNA hybridization, electrophoretic migration of malate and glutamate dehydrogenase, and peptidase and lipase activities of type strains. Gas chromatography of cellular fatty acids can be a useful adjunct for characterization and identification of bacterial species. In the present study, cellular fatty acid profiles were determined for seven strains of P. intermedia and six strains of P. nigrescens. Six of these 13 strains were isolated from the root canal and blood of three patients during endodontic therapy of teeth with Asymptomatic apical periodontitis. The bacteria were cultivated anaerobically in 10 mL prereduced anaerobically sterilized peptone-yeast extract-glucose broth for 24 h. Dried cells of each isolate were methanolysed and their fatty acid contents determined by the Microbial Identification System software package by MIDI. The data were treated by principal component analysis, which distinguished P. nigrescensfromP. intermedia. Cellular fatty acid profiles of these strains of the species in blood matched the profiles of their respective root canal isolates, as demonstrated by Euclidean Distance Square assessment. This suggested that the organisms in the root canal had spread to the bloodstream during endodontic treatment.     

Growth of oral Streptococcus species and Actinomyces viscosus in human saliva.

Microorganisms in dental plaque live in constant association with saliva. The role of saliva in the adherence of bacteria to the teeth and the antibacterial properties of saliva have been well investigated; less interest has been shown in the possible role of saliva as a substrate for oral microorganisms. In this study it was shown that saliva can serve as a growth medium for oral Streptococcus spp. and Actinomyces viscosus. The cell production of these organisms on saliva was carbohydrate limited. The doubling times for growth on glucose-supplemented saliva (4 to 5 mmol/liter) ranged from 1.6 to 4.0 h. The availability of carbohydrate sources for the oral microflora is discussed in relation to microbial growth in the oral cavity.     

2种不同器械扩锉后根管壁的细菌学研究

目的通过对临床病例的研究,比较减速镍钛机扩和普通不锈钢K型锉清洁根管的能力。方法选取临床需要根管治疗的患牙60例,随机分为2组,经开髓拨髓后分别用镍钛减速机扩和不锈钢K型及H型锉进行根管预备,预备完成后用灭菌棉捻擦拭管壁,作微生物检查。结果2组方法扩锉后对根管内微生物的变化做统计学分析,差异有显著性(P<0.05)。结论镍钛减速机扩对根管的清洁能力高于普通不锈钢扩锉,值得临床的推广。     

脉冲Nd∶YAG激光在根管预备中的临床应用——与超声根管预备、传统手动根管预备比较

为了观察脉冲Ｎｄ∶ＹＡＧ激光在根管预备（以下简称根备）中的作用,我们用Ｎｄ∶ＹＡＧ激光辅助,对３０颗患牙进行根备,根备后即时根充。并与超声根备后即时根充、传统根备后即时根充比较,结果表明：激光组的术后疼痛反应少于超声组与传统组,远期疗效与超声组相仿,而优于传统组。     

新型激光光敏根管消毒剂在牙本质中渗透性的研究

目的：观察新型激光光敏根管消毒剂、甲苯胺蓝和亚甲基蓝在牙本质中的渗透效果,评价三种光敏剂在牙本质中的渗透性。方法：收集新鲜拔除的离体单根管牙90颗,根管预备后随机分为三组。每组30颗。A组为新型激光光敏根管消毒剂组;B组为甲苯胺蓝组;c组为亚甲基蓝组。A、B、C三组实验根管内分别用浸有饱和的新型激光光敏根管消毒剂、甲苯胺蓝和亚甲基蓝的棉捻在根管内停留60秒。沿牙体长轴颊舌纵向劈开,倒置荧光显微镜下观察并拍照,用Photoshop8．01软件测量三种光敏剂渗透入牙本质的平均深度。结果：新型激光光敏根管消毒剂在牙本质内的渗透平均深度为553．25μm,甲苯胺蓝在牙本质内的渗透平均深度为350．75μm,亚甲基蓝在牙本质内的渗透平均深度为168．25μm。结论：新型激光光敏根管消毒剂在牙本质渗透性明显优于甲苯胺蓝和亚甲基蓝,具有良好的牙本质渗透性。     

儿童乳牙根管感染的细菌学分析

对18例3～8岁儿童乳牙的根管感染以无菌技术进行定量取样,按种于12种选择性培养基和2种非选择性培养基上,进行需氧、微需氧和厌氧培养,并对细菌菌落计数。对牙髓拟杆菌和牙龈拟杆菌作半定量免疫荧光染色计数;并对其中9例病牙进行了菌相分析。检出的所有细菌中,厌氧菌占绝对优势;其中检出率较高的菌为:产黑色素拟杆菌属,厌氧革兰氏阳性球菌,微需氧革兰氏阳性球菌等.本试验证明,儿童乳牙根管感染是以厌氧菌为主的混合感染,其中以产黑色素拟杆菌属等最常见.     

三种根管充填材料对 乳牙根尖周炎常见细菌的抑菌性

目的评估和比较3种乳牙根管充填材料对乳牙根尖周炎常见细菌的抗菌功效。方法在体外实验中,采用纸片扩散法检测并观察3种乳牙根管充填材料及蒸馏水作用于粪肠球菌、牙龈卟啉单胞菌和具核梭杆菌后形成的抑菌环直径(mm),用SPSS 24.0软件进行统计分析。结果三重抗生素糊剂对乳牙根尖周炎根管中3种细菌均显示出最强的抗菌潜力,其次是Vitapex?、氢氧化钙糊剂,结果差异具有统计学意义(P0.05);蒸馏水对3种细菌是非抑制性的。粪肠球菌对3种药物反应最敏感,其次是牙龈卟啉单胞菌、具核梭杆菌,结果差异具有统计学意义(P0.05)。结论三重抗生素糊剂是最推荐用于治疗乳牙根尖周炎的根管充填材料。     

联合方法测量工作长度应用于颌骨囊肿受累牙根管治疗的研究

目的：研究颌骨囊肿受累牙根管治疗中采用联合方法测量根管工作长度的效果。方法：选取36 例颌骨囊肿受累患牙需术前 根管治疗的患者,共135 颗患牙,186 个根管,随机分为两组,实验组（93 个根管）采用电测法联合纸尖法测量根管工作长度,对照 组（93 个根管）则采用电测法联合X 线法测量根管工作长度。对比两组根管充填后的充填效果。结果：实验组的根管恰填率为 90.32%,高于对照组的79.57%（P＜0.05）。结论：采用电测法联合纸尖法测量颌骨囊肿受累牙根管工作长度,准确率高,根管充填 效果好且安全。     

Evaluation of the strain identity between isolates from caries lesions and root canals in early childhood caries cases

Early childhood caries (ECC) has become a serious medical problem worldwide in the last decade. Bacterial microflora of the dental plaque and oral cavity is considered an important factor in the formation and progression of dental caries. The aim of this study was strain typing and comparison of bacterial isolates retrieved from caries lesions and root canal contents of the same teeth. In total, 18 pairs of presumptive streptococci and lactobacilli retrieved from dental caries and root canals isolated from ECC-affected children, were selected on the basis of biotyping results and rep-PCR fingerprinting with (GTG)₅ primer. Strain typing was further done using the RiboPrinter microbial characterization system (DuPont Qualicon). The automated ribotyping determined 14 pairs of the strains (77.8 %) to be identical. The results obtained confirmed that identical bacterial strains colonized both the decayed dental surface and the necrotic content of the dental pulp cavity during the cariogenesis. Our finding supports the assumption that bacteria could penetrate through the damaged dental surface to the inner parts of the teeth.     

Expression of the short fimbriae of Porphyromonas gingivalis is regulated in oral bacterial consortia

The Mfa1 protein of Porphyromonas gingivalis is the structural subunit of the short fimbriae and mediates coadhesion between P. gingivalis and Streptococcus gordonii. We utilized a promoter-lacZ reporter construct to examine the regulation of mfa1 expression in consortia with common oral plaque bacteria. Promoter activity of mfa1 was inhibited by S. gordonii, Streptococcus sanguinis and Streptococcus mitis. In contrast, Streptococcus mutans, Streptococcus cristatus, Actinomyces naeslundii, Actinobacillus actinomycetemcomitans and Fusobacterium nucleatum did not affect mfa1 expression. Expression of SspA/B, the streptococcal receptor for Mfa1, was not required for regulation of mfa1 promoter activity. Proteinaceous molecule(s) in oral streptococci may be responsible for regulation of Mfa1 expression. Porphyromonas gingivalis is capable of detecting heterologous organisms, and responds to selected organisms by specific gene regulation.     

糖尿病患者拔牙创区微生物分布的临床研究

探讨糖尿病患者拔牙术后创区微生物分布情况 ,为临床防治术后感染提供理论依据。随机选取 3 9例已确诊为糖尿病的拔牙患者 ,在术后即刻采取拔牙创的渗出物 ,按 1 0倍系列稀释 ,培养微生物 ,然后进行分离、纯化和鉴定菌。结果从标本中培养出 1 87株微生物 ,鉴定到属或种 ,共获得 1 1种不同的细菌 ,常见的 7种。研究结果表明 ,糖尿病患者拔牙创区常见微生物依次为链球菌、萘瑟氏菌、梭杆菌、产黑色素拟杆菌、拟杆菌 ,韦荣菌属、二氧化碳嗜纤维菌属等。其中 ,链球菌、萘瑟氏菌为优势菌。     

The conditioning role of saliva in streptococcal attachment to hydroxyapatite surfaces

The solution properties of saliva and its role in conditioning both the substrate and the bacterial surface have been investigated with regard to the attachment of oral streptococci to hydroxyapatite surfaces. Saliva from eight subjects was used and the attachment of three organisms, Streptococcus mutans strains FA-1 (serotype b) and KPSK2 (serotype c) and S. sanguis T175-1, was studied. An adsorbed salivary layer on a hydroxyapatite surface substantially reduced the affinity of the organisms for the surface. Adsorbed saliva on the bacterial surfaces, however, tended to increase the organisms' affinity for saliva-coated apatite. The source of saliva was important in determining the extent of inhibition of attachment. The data indicated that the negatively charged and hydrophilic nature of salivary conditioning films was important in controlling bacterial adsorption to hydroxyapatite. The results also suggested that hydrophobic salivas could promote binding of the more hydrophobic bacteria known to be early colonizers of the teeth.     

速效牙痛宁酊辅助根管治疗对慢性牙髓炎患者口腔健康相关生活质量和血清Toll样受体4、CX3C趋化因子配体1的影响

摘要 目的：观察速效牙痛宁酊辅助根管治疗对慢性牙髓炎患者口腔健康相关生活质量和血清CX3C趋化因子配体1（CX3CL1）、Toll样受体4（TLR4）的影响。方法：选取衡水市中医院2019年9月~2021年6月期间收治的87例慢性牙髓炎患者（患牙87颗）,根据随机数字表法分为观察组44例（患牙44颗,给予速效牙痛宁酊辅助根管治疗）、对照组43例（患牙43颗,给予根管治疗）。对比两组临床疗效,观察两组疼痛症状和口腔健康相关生活质量改善情况,比较两组血清TLR4、CX3CL1水平变化,并记录两组不良反应发生情况。结果：观察组的临床总有效率高于对照组（P<0.05）。治疗2周后、4周后,两组视觉模拟量表（VAS）评分降低,且观察组较对照组低（P<0.05）。治疗4周后,两组口腔健康影响程度量表（OHIP-14）各领域（社交障碍、心理障碍、功能限制、生理障碍、残障、生理疼痛、心理不适）评分及总分降低,且观察组低于对照组（P<0.05）。治疗4周后,两组血清CX3CL1、TLR4水平下降,观察组较对照组低（P<0.05）。与对照组比较,观察组并发症总发生率较低（P<0.05）。结论：速效牙痛宁酊辅助根管治疗慢性牙髓炎患者,可有效减轻疼痛症状,提高口腔健康相关生活质量,作用机制可能与调节血清TLR4、CX3CL1水平有关。