牙周炎患者龈下菌斑菌群的高通量测序分析

目的利用高通量测序(high?throughput sequencing,HTS)分析广泛型侵袭性牙周炎(generalized ag?gressive periodontitis,GAgP)和重度慢性牙周炎(severe chronic periodontitis,SCP)龈下菌群的组成,利用生物信息学分析其多样性及功能,并观察牙周基础治疗前后龈下菌群的变化。方法选择2018年9月至2019年5月在昆明医科大学附属口腔医院牙周病科就诊的11例GAgP患者和14例SCP患者作为研究对象。在基线及牙周基础治疗后第6周采集龈下菌斑样本并提取DNA,进行MiSeq测序,采用QIIME(quantitative insights in mi?crobial ecology)、Mothur及SPSS等软件分析群落信息,并进行LEfSe差异分析(linear discriminant analysis Effect size,LEfSe)、Network网络分析,使用KEGG PATHWAY数据库(https://www.kegg.jp/kegg/pathway.html)对群落功能进行预测。结果基线时,GAgP和SCP患者的优势菌群相似,均包括拟杆菌门、卟啉单胞菌属和牙髓卟啉单胞菌等。治疗后第6周,GAgP和SCP患者菌群变化相似,由于牙周袋变浅,拟杆菌门、卟啉单胞菌属和牙髓卟啉单胞菌等革兰阴性菌相对丰度降低,变形菌门、放线菌属和空间罗氏菌等革兰阳性菌相对丰度增加;其中放线菌是GAgP治疗后龈下菌群中显著增加的生物标志物;链球菌属是联系牙周炎相关菌属和牙周健康相关菌属的重要菌属。群落功能预测表明基础治疗可降低GAgP和SCP患者群落中的氨基酸代谢、甲烷代谢和肽酶等功能。结论GAgP和SCP患者的龈下菌群相似,链球菌属作为早期定植菌,在龈下菌群由健康向失调转变的过程中可能发挥着促使菌斑生物膜形成及成熟的重要作用;牙周基础治疗可以改变GAgP和SCP患者龈下菌群的组成和结构,降低群落多样性,降低氨基酸代谢、甲烷代谢和肽酶等群落功能。     

Nd: YAP激光治疗牙周炎的疗效评价及其对龈下微生物的影响

目的 比较龈下刮治和根面平整术(scaling and root planning, SRP)联合Nd:YAP激光与单纯SRP治疗对牙周炎患者疗效的差异和龈下微生物的影响。方法 选择符合纳入标准的牙周炎患者。每位患者龈上洁治1周后为本研究基线,随机半口分组。试验侧采用SRP联合Nd: YAP激光治疗;对照侧仅采用SRP治疗。每位患者选择一组同颌同名单根牙,进行龈沟液(gingival crevicular fluid, GCF)和龈下菌斑取样。分别在基线、治疗完成后6周和12周进行探诊深度(probing depth, PD)和探诊出血(bleeding on probing, BOP)的数据采集、龈沟液及龈下菌斑样本的采集。ELISA法检测龈沟液中炎症因子白细胞介素6(interleukin-6,IL-6)的浓度。16S rDNA高通量测序对各龈下菌斑样本的菌群构成进行分析和比较。结果 试验侧与对照侧的各项临床指标较基线时均有显著改善(P＜0.05),但两组间无明显差异。两组的IL-6浓度较基线均显著下降,治疗后6周,试验侧的 IL-6浓度明显低于对照组(P＜0.05)。龈下菌斑α多样性分析,治疗后6周及12周,对照侧的Shannon指数高于基线,Simpson指数低于基线(P＜0.05)。测序结果显示,治疗后6周,试验侧的普氏菌属(Prevotella)和Saccharibacteria_incertae_incerta_sedis的相对丰度较基线显著下降;密螺旋体属(Treponema)的相对丰度低于对照侧,嗜二氧化碳噬细胞菌属(Capnocytophaga)的相对丰度高于对照侧(P＜0.05),12周时无明显差异。结论 在本研究中,Nd:YAP激光联合SRP和单纯SRP治疗牙周炎均有明显效果,而两种治疗方法在临床指标上无明显差异,辅助使用Nd:YAP激光在短时间内更有利于GCF IL-6浓度的降低和部分牙周致病菌的控制。     

光动力疗法结合龈下喷砂对2型糖尿病牙周炎的临床疗效研究

目的:探究光动力疗法(photodynamic therapy, PDT)结合龈下喷砂对2型糖尿病(type 2 diabetes mellitus, T2DM)合并牙周炎患者的临床治疗效果。方法:选取2021年7月～2023年1月本院收治的82例2型糖尿病伴牙周炎患者,随机均分为对照组(n=41)和研究组(n=41),对照组给予牙周基础治疗,研究组则加用光动力疗法结合龈下喷砂,对比治疗前和治疗后3个月患者牙周临床指标、龈沟液细胞因子水平和龈下菌斑菌群结构的变化。结果:治疗后,研究组出血指数(bleeding index, BI)、探诊深度(probing depth, PD)和临床附着丧失(clinical attachment loss, CAL)低于对照组(P<0.05);C反应蛋白(C reactive protein, CRP)、抵抗素低于对照组(P<0.05),超氧化物歧化酶(superoxide dismutase, SOD)高于对照组(P<0.05)。菌群分析显示治疗3个月后研究组变形菌门、拟杆菌门相对丰度低于对照组(P<0.05),厚壁菌门相对...     

超声和手工龈下刮治对龈下微生物影响的研究

目的 :比较Perioproline超声龈下刮治和手工龈下刮治对龈下微生物的影响 ,评价Periopro line超声龈下刮治系统的临床疗效和工作效率。方法 :选取慢性成人牙周炎患者 7名 ,共 2 0个左右基本对称牙被纳入研究 ,随机分入实验组 (超声组 )和对照组 (手工组 )。记录治疗时间和治疗前、治疗后 0d、7d和 14d的PD ,BOP值 ,及采集龈下菌斑样本培养鉴定 ,观察比较总菌落形成单位 (CFU)和牙周可疑致病菌检出率及检出量的变化。结果 :两组在治疗后PD、BOP值及龈下菌斑总量 (CFU)和牙周可疑致病菌数量及检出率均显著下降 (P <0 .0 5) ;治疗前后各时段组间均无明显差别 ;超声组的治疗时间明显少于手工组 (P <0 .0 5)。结论 :Perioproline超声龈下刮治和手工龈下刮治均可有效减少龈下微生物的量 ,改善患牙的牙周临床状况 ,且两者间无差异 ,但Perioproline超声龈下刮治明显较手工刮治节省时间     

Ⅱ型糖尿病合并牙周炎患者龈下菌斑微生物群落分析

目的：本研究旨在比较Ⅱ型糖尿病合并慢性牙周炎患者与无糖尿病的牙周炎患者龈下菌斑微生物构成。方法：12名患者分为糖尿病合并慢性牙周炎组（T2DM＋CP组）与慢性牙周炎组（CP组）2组,各6人。记录所有患者基本信息及牙周临床参数,包括年龄、性别、探诊深度和附着丧失。根据探诊深度和附着丧失取患病位点的菌斑样本。PCR检测7种牙周可疑致病菌。采用DGGE分离扩增的16SrDNA片段。结果：两组结果显示两组牙周参数无显著差异。两组7种细菌检出率相似。所有对象中均检出牙龈卟啉单胞菌、福塞坦氏菌、齿垢密螺旋体和中间普氏菌,而具核梭杆菌在两组中均有一个样本未检出。变黑普氏菌在T2DM＋CP组的2个样本中检出,而CP组有4个样本检出。伴放线共聚菌在所有样本中均未检测到。DGGE分析结果示两组间条带数量及树状聚类分析均无显著差异。结论：Ⅱ型糖尿病合并牙周炎患者龈下菌斑的牙周可疑致病菌的检出情况以及DGGE分析与无糖尿病患者相似。     

伴放线放线杆菌在龈下菌斑和颊黏膜分布研究

目的:比较伴放线放线杆菌(actinobac illus actinomycetem com itans,A.a)在不同类型牙周炎患者龈下菌斑和颊黏膜中的分布。方法:通过聚合酶链反应(polym erase chain reaction,PCR)对侵袭性牙周炎患者(AgP)、慢性牙周炎患者(CP)、牙周健康者口腔龈下菌斑和颊黏膜中的A.a进行检测,分析该菌分别在两部位的相对含量。结果:AgP组菌斑和颊黏膜样本中A.a阳性检出率均为41.7%,分别高于CP组(菌斑16.7%、颊黏膜10.0%)和牙周健康组(菌斑和颊黏膜均为0%)。AgP组A.a在菌斑和颊黏膜的相对含量分别为38.5%和22.2%,高于CP组(菌斑19%、颊黏膜12.75%)。结论:A.a不仅存在于龈下菌斑中,也能够粘附于颊黏膜;A.a是AgP的主要优势菌也参与了CP的菌群组成。     

牙周致病菌在冠心病患者龈下菌斑中的分布

目的:分析牙周致病菌在冠心病患者龈下菌斑中的分布情况,及冠心病患者龈下菌斑中牙周致病菌的分布与慢性牙周炎(chronic periodontitis,CP)病变程度之间的关系。方法:收集44例患冠心病并伴有CP患者的龈下菌斑,采用Chelex-100法提取细菌DNA,以聚合酶链反应(polymerase chain reaction,PCR)法检测龈下菌斑中牙龈卟啉单胞菌(Porphyromonas gingivalis,Pg)、伴放线菌嗜血菌(Haemophilus actinomycetemcomitans,Ha)、具核梭杆菌(Fusobacterium nucleatum,Fn)、中间普氏菌(Prevotella intermedia,Pi)、福赛斯坦纳菌(Tannerella forsythensis,Tf)五种牙周炎相关致病菌。结果:44例冠心病患者的龈下菌斑中各牙周致病菌的检出率分别为:Pg 19(43.18%)、Ha 9(20.45%)、Pi 27(61.36%)、Fn 38(86.36%)、Tf 41(93.18%);轻度CP患者1例,Fn 1例(100%);中度CP患9例,其中Pg6(66.67%)、Ha 2(22.22%)、Pi 7(77.78%)、Fn 9(100%)、Tf 8(88.89%);重度CP患者34例,其中Pg13(38.24%)、Ha 7(20.59%)、Pi 20(58.82%)、Fn 28(82.35%)、Tf 33(97%)。结论:Tf、Fn可能在冠心病的发生发展中起着重要作用,冠心病患者中CP的发病和进展可能有其特定的细菌学病因。     

慢性牙周炎龈下菌斑和唾液微生物群落分析

目的:运用DGGE技术检测慢性牙周炎龈下菌斑和唾液微生物群落结构,分析其菌群之间的关系。方法:选取34名慢性牙周炎志愿者,分别采集龈下菌斑和唾液样本,采用PCR扩增细菌16Sr RNA V3区后,运用DGGE技术分析各组样本微生物群落结构。通过数字化软件Im age J将DGGE凝胶图谱转成数字信息,对所得矩阵进行主成分分析(Princ ipal ComponentAnalysis,PCA)、聚类分析(C luster Analysis)和偏最小二乘法(Partial Least Squares,PLS)分析。结果:DGGE图谱显示:龈下菌斑DGGE条带数目为11～28,,平均19;唾液DGGE条带数目为11～24,平均17;PCA结果显示:龈下菌斑与唾液比较有明显的差异;聚类分析显示:龈下菌斑、唾液形成5个明显聚类群,在同一聚类群中相似度高,在不同聚类群中相似度低,表示龈下菌斑与唾液菌群之间有显著差异;PLS结果显示:慢性牙周炎龈下菌斑与唾液菌群有显著差异。结论:DGGE是一种能直观显示微生物群落的指纹技术。通过该技术能检测牙周、唾液微生物群落的结构和组成,本结果显示,慢性牙周炎龈下菌斑与唾液微生物群落结构有明显差异。     

Er: YAG激光对2型糖尿病相关性牙周炎患者龈下菌群动态变化的研究

目的 动态观察2型糖尿病相关性牙周炎患者经Er: YAG激光治疗前后龈下菌群的变化,并与不伴全身疾病的慢性牙周炎龈下菌群进行比较。方法 收集11例2型糖尿病相关性牙周炎患者的13对患牙（26个位点）作为试验组,分别进行Er: YAG激光治疗和超声治疗,采集治疗前及治疗后1、3个月的龈下菌斑;同时收集11例牙周状况相近的不伴全身疾病的中重度慢性牙周炎患者13个位点的龈下菌斑作为对照组,分析试验组是否有菌群种类变化。提取龈下菌斑DNA,进行变性梯度凝胶电泳分离及条带回收测序。结果 试验组与对照组患牙龈下菌斑的优势致病菌存在差异,分别为中间普雷沃菌和福赛斯坦纳菌。激光组治疗前后龈下菌群构成也发生改变,治疗后1个月,有的条带表达减弱或消失,并有新条带出现;测序结果表明,新出现的条带为放线菌的一种,减弱、消失的为福赛斯坦纳菌。结论 Er: YAG激光治疗前后龈下菌群的构成发生变化,治疗后1个月是关键时期;治疗后3个月,激光治疗在阻止细菌再定植方面可能更具优势。     

不同牙周状况龈下菌斑中福赛斯坦纳菌分布

目的：分析牙周健康者和慢性牙周炎病人龈下菌斑中福赛斯坦纳菌的分布情况,探讨该菌与不同牙周状况的关系。方法：收集111例牙周健康者、108例慢性牙周炎病人,病变位点和健康位点共327个龈下菌斑,采用16S rRNA PCR方法检测福赛斯坦纳菌的分布。结果：牙周炎病人病变位点福赛斯坦纳菌检出率为56.5%,显著高于牙周健康者（10.8%,P〈0.001）。慢性牙周炎病人健康位点福赛斯坦纳菌检出率为3.7%,显著低于病变位点（P〈0.001）,牙周健康者该菌的检出率比牙周炎健康位点高（P〈0.05）。结论：龈下菌斑中福赛斯坦纳菌的存在与牙周炎症的发生有密切关系。     

光动力疗法对慢性牙周炎龈下牙周致病菌的影响

目的：应用PeriowaveTM光动力杀菌系统对慢性牙周炎患者进行治疗,通过龈下菌斑中牙周致病菌比例的变化,评价光动力疗法（photodynamic therapy,PDT）治疗慢性牙周炎的临床效果。方法：选取60名慢性牙周炎患者,分别给予SRP＋1次PDT（A组）、SRP＋2次PDT（B组）或单纯SRP（C组）治疗。利用real-time PCR技术检测A、B、C三组在治疗前、治疗后6周、治疗后12周龈下菌斑中牙周致病菌P.g、A.a、T.f所占比例的变化。结果：治疗后6周,A、B两组牙周致病菌P.g在总菌中的比例都有非常显著降低（p〈0.01）,C组有显著降低（p〈0.05）;治疗后12周,A、B组仍有非常显著的降低（p〈0.01）,与C组相比有非常显著差异（p〈0.01）,但A、B两组之间没有差异（p〉0.05）;而仅在治疗后12周,B组的A.a相对于总菌的比例与基线相比有显著降低（p〈0.05）,且这一变化显著大于A、C两组（p〈0.05）;在治疗后6周,A、B组T.f相对于总菌含量的下降与基线相比,明显大于C组（p〈0.05）,治疗后12周,A、B两组T.f相对于总菌的含量仍有非常显著的下降（p〈0.01）,但A组和B组之间都没有差异。结论：PDT对P.g、A.a和T.f3种牙周致病菌都有杀灭作用,但P.g、T.f对PDT更为敏感,PDT可以作为治疗慢性牙周炎的辅助方法。     

甘氨酸龈下喷砂辅助治疗老年伴糖尿病牙周炎疗效分析

目的:观察甘氨酸龈下喷砂辅助治疗老年伴糖尿病牙周炎后,牙周炎症的改善情况以及糖尿病相关指标的变化.方法:2018年6月～2019年6月于浙江医院口腔科就诊的40例老年伴糖尿病牙周炎患者(男20例,女20例,年龄60～78岁)为研究对象.随机分为对照组(超声龈下刮治+根面平整术)和实验组(超声龈下刮治+根面平整术+甘氨酸...     

福赛斯坦纳菌在2型糖尿病患者龈下菌斑中的分布

目的 观察福赛斯坦纳菌(T.forsythensis)在2型糖尿病患者不同牙周状况下的分布及其与相关影响因素的关系,探讨T.forsythensis在慢性牙周炎与2型糖尿病中的作用.方法采集160例不同牙周状况的2型糖尿病患者的龈下集合菌斑,采用酚-氯仿法提取细菌DNA,应用聚合酶链反应法对T.forsythensis...     

Description of the subgingival microbiota of periodontally untreated Mexican subjects: chronic periodontitis and periodontal health

BACKGROUND: Recent studies have suggested that changes in the prevalence and/or proportion of distinct microorganisms characterize the subgingival microbial profiles of populations around the world. At present, no information is available on the subgingival microbiota of Mexican subjects. The purpose of the present study was to determine the microbial composition of subgingival plaque in Mexican subjects with untreated chronic periodontitis. METHODS: A total of 44 chronic periodontitis and 20 periodontally healthy subjects (who were currently non-smokers) were selected. Clinical measurements including plaque accumulation, gingival erythema, bleeding on probing, suppuration, probing depth, and attachment level were recorded at six sites of every tooth. Up to 28 subgingival plaque samples were obtained from each subject and individually analyzed to determine the levels, proportion, and prevalence of 40 microbial species using the checkerboard DNA-DNA hybridization technique. RESULTS: Porphyromonas gingivalis, Treponema denticola, and Tannerella forsythensis were the only species that presented higher mean levels in periodontitis subjects. The proportions of P. gingivalis (P<0.001), T. forsythensis (P<0.01), and red complex species (P. gingivalis, T. forsythensis, and T. denticola; P<0.001) as a group were also significantly higher in periodontitis subjects. Periodontally healthy subjects harbored a significantly larger proportion of Actinomyces species (P<0.05). No significant differences were detected in the percentage of carriers of any of the species tested. CONCLUSIONS: Our results revealed that the subgingival microbiota of untreated chronic periodontitis Mexican subjects was characterized by increases in the level, prevalence, and proportion of classic periodontal pathogens. However, the prevalence and proportion of specific microbial species varied significantly from the results of other reports on subjects from different geographical locations.     

Non-surgical periodontal therapy with adjunctive topical doxycycline: a double-masked, randomized, controlled multicenter study. II. Microbiological results

BACKGROUND: Topical application of active substances offers an additional option in periodontal therapy. The aim of this study was to examine the influence of the administration of a novel, biodegradable 14% doxycycline gel on microbiological findings, in connection with scaling and root planing. METHODS: One hundred ten patients in three centers (Frankfurt and Heidelberg, Germany; Nijmegen, The Netherlands) with moderate to advanced periodontitis were evaluated in this randomized, double-masked, split-mouth clinical trial. In each patient, three test teeth were randomly assigned to one of three treatment modalities: 1) scaling and root planing (SRP) alone; 2) SRP with subgingival placebo gel (VEH); or 3) SRP and 14% doxycycline gel (DOX). Subgingival plaque samples were analyzed at baseline and 3 and 6 months after therapy for Actinobacillus actinomycetemcomitans (A.a.), Tannerella forsythensis (T.f.), Porphyromonas gingivalis (P.g.), and Treponema denticola (T.d.) using a RNA probe. Samples from 10 patients were tested for resistance against doxycycline, amoxicillin/clavulanic acid, cefoxitin, clindamycin, and metronidazole using agar diffusion testing. RESULTS: The largest decrease in pathogens was found after 3 months, with the most pronounced differences between DOX and SRP (P <0.05). At 6 months, pathogens were still reduced markedly in all groups. Treatment results were consolidated for VEH and DOX, with a slight deterioration for SRP (DOX versus SRP: P <0.001). Resistance was observed to amoxycillin/clavulanic acid, cefoxitin, clindamycin, and metronidazole (four isolates) but not to doxycycline. CONCLUSION: The addition of subgingival instillation of a 14% doxycycline gel resulted in pronounced reduction of periodontal pathogens after 3 months and stabilizing results up to 6 months after therapy. Resistance to doxycycline was not induced.     

Clinical and microbiological effect of scaling and root planing in smoker and non-smoker chronic and aggressive periodontitis patients

OBJECTIVES: To compare the effects of scaling and root planing (SRP) on clinical and microbiological parameters at selected sites in smoker and non-smoker chronic and generalized aggressive periodontitis patients. MATERIALS AND METHODS: Clinical parameters including probing depth (PD), relative attachment level (RAL), and bleeding upon probing (BOP), and subgingival plaque samples were taken from four sites in 28 chronic periodontitis (CP) and 17 generalized aggressive periodontitis (GAgP) patients before and after SRP. Polymerase chain reaction assays were used to determine the presence of A. actinomycetemcomitans, Porphyromonas gingivalis, Tannerella forsythensis, Prevotella intermedia and Treponema denticola. RESULTS: Both CP and GAgP non-smokers had significantly greater reduction in pocket depth (1.0+/-1.3 mm in CP smokers versus 1.7+/-1.4 mm in non-smokers, p=0.007 and 1.3+/-1.0 in GAgP smokers versus 2.4+/-1.2 mm in GAgP non-smokers, p<0.001) than respective non-smokers, with a significant decrease in Tannerella forsythensis in CP sites (smokers 25% increase and non-smokers 36.3% decrease, p<0.001) and Prevotella intermedia at GAgP sites (smokers 25% reduction versus 46.9% in non-smokers, p=0.028). CONCLUSION: SRP was effective in reducing clinical parameters in both groups. The inferior improvement in PD following therapy for smokers may reflect the systemic effects of smoking on the host response and the healing process. The lesser reduction in microflora and greater post-therapy prevalence of organisms may reflect the deeper pockets seen in smokers and poorer clearance of the organisms. These detrimental consequences for smokers appear consistent in both aggressive and CP.     

单纯机械治疗对慢性牙周炎患者龈下菌群微生态的影响

目的 采用16S rRNA高通量测序技术,探讨单纯机械治疗对慢性牙周炎龈下菌群微生态的影响.方法 纳入广泛型中重度慢性牙周炎患者,接受口腔卫生宣教,并同时进行龈上洁治.一周后进行超声结合手工龈下刮治及根面平整,在基线、治疗后3个月和治疗后6个月,记录临床指标并采集龈下菌斑.利用Illu-mina MiSeq平台进行16...     

Effects of metronidazole plus amoxicillin as the only therapy on the microbiological and clinical parameters of untreated chronic periodontitis

AIM: To determine the effect of metronidazole plus amoxicillin (M+A) as the sole therapy, on the subgingival microbiota of chronic periodontitis. MATERIAL AND METHODS: Twenty-two patients with untreated chronic periodontitis were randomly assigned to a group that received M+A for 7 days, or to a group receiving scaling and root planing (SRP) and two placebos. Clinical measurements including sites with plaque, bleeding on probing (BOP), probing depth (PD) and attachment level (AL) were made at baseline, 3, 6, 9 and 12 months. Subgingival plaque samples were taken from all teeth at baseline 3, 6, 9 and 12 months for the counts of 40 subgingival species using checkerboard DNA-DNA hybridization. RESULTS: Mean PD was reduced from 2.80+/-0.45 at baseline to 1.95+/-0.05 at 12 months (P<0.001) and from 2.39+/-0.41 to 1.95+/-0.10 (P<0.001) in the M+A- and SRP-treated patients, respectively. Corresponding values for relative mean AL were 10.07+/-1.30-9.77+/-0.34 (P<0.001) and 9.94+/-0.28-9.77+/-0.26 (P<0.001). Percentage of sites exhibiting BOP were 40.6+/-18.3-14.0+/-1.4 (P<0.001), and 38.5+/-5.1-19.0+/-2.8 (P<0.001) in the M+A and SRP groups, respectively. Mean total DNA probe counts and counts of the majority of the 40 test species were significantly reduced over time in both groups, with no significant differences detected at any time point between groups. At 12 months many of the species were still present at significantly lowered levels compared with their baseline counts in both groups. CONCLUSIONS: Changes in clinical and microbiological parameters were similar after receiving systemically administered M+A as the sole therapy or after receiving SRP only.