染色法检测与牙周病相关的龈下细菌

牙周炎时,龈下菌斑细菌组成的检查是判定牙周病活跃性很重要的辅助手段。当前已广泛使用暗视野显微镜检测龈下微生物,特别是对螺旋体和可动菌的检测。相差显微镜也用于此目的,但不如暗视野检查法清晰。暗视野显微镜检法和褶差显微镜检法独特之处是可在一张片子上同时观察标本的螺旋体和可动菌,是其它方法不能比拟的。但是,暗视野显微镜的     

两种椅旁显微镜检查方法的比较

对17名牙周炎患者的85份龈下菌斑样本,用暗视野显微镜检查法和刚果红涂片染色法,进行方法学的比较,探讨它们的优缺点和应用中的注意问题。结果表明:两种方法对球菌、螺旋体和其它细菌的检查无显著性差异,此两种方法对以形态学为基础的微生物分类检查具有平行意义,两法各有其优缺点,如同时应用,可以互相补充。     

龋齿、牙周炎患牙和健康牙的菌斑生物膜特征

目的研究牙面菌斑生物膜特征与口腔疾病的关系。方法选择牙周健康而牙冠严重龋坏的龋齿5颗、无龋损而极度松动的牙周炎患牙6颗及正畸原因拔除的健康牙4颗,在扫描电镜下,观察分析龈上、龈下及移行生态区的菌斑生物膜特征。结果龋齿、牙周炎患牙和健康牙的牙面均观察到细菌混合物组成的菌斑生物膜,健康牙菌斑生物膜以球菌为主,放线菌和短杆菌少量;龋齿牙的龋坏处为坏死组织和细菌,龋边缘及龈沟处的球菌和短杆菌较健康牙多;牙周炎患者牙菌斑生物膜的细菌种类多,在龈上、龈下移行处可见典型的玉米棒状菌斑或以杆菌为主的紧密附着菌斑,龈下可见球菌、杆菌、梭菌及螺旋体等构成的复杂菌斑。结论龋齿、牙周炎患牙和健康牙菌斑生物膜细菌组成、集聚秩序和立体结构不同,菌斑生物膜的形成与细菌的附着、集聚、生长有关,也与局部病变密切相关。     

纳米TiO2陶瓷涂层托槽周边菌斑附着研究

目的：评估纳米TiO2陶瓷涂层托槽周边菌斑附着情况。方法：选择20例正畸拔牙矫治病例,于待拔除前磨牙粘接自行研制的纳米TiO2陶瓷涂层直丝弓托槽。托槽粘接后1周、2周和3周拔除测试牙,电镜扫描观察托槽周边区域、托槽龈方区域菌斑附着。结果：①托槽周边区域存在大量多余粘接剂,表面粗糙且粘接剂．釉质间存在明显间隙;②大量菌斑附着,托槽龈方区域清洁困难,菌斑附着较多;③托槽粘接后2周、3周时,托槽周边区域粘接剂表面菌斑成熟,而托槽龈方近牙颈部区域菌斑尚处于形成初期。结论：托槽粘接后,托槽周边区域及龈方近牙颈部区域是菌斑聚集的主要区域。     

控制菌斑防治牙龈炎100例疗效观察

通过临床研究,控制菌斑防治龈炎是有效的,现报道如下。1材料和方法1.1病例选择为我院牙周病门诊患者100例(男78,女22),年龄20~50岁,随机将患者分为实验组和对照组,每组各50例。两组在职业、性别、年龄上无明显差异。1.2临床各项观察指标按O′Leary的菌斑控制记录卡计算菌斑指数。按L e(1967)提出的标准,将龈炎分为4级。1.3微生物学检查取患者龈下菌斑和少许分泌物,混匀于事先备好的20 g/L刚果红溶液玻片中,均匀推片干燥后,用盐酸熏蒸至变为蓝色,在高倍显微镜下计数200个微生物球菌、杆菌、螺旋体比例。1.4实验方法和统计学处理将实验组和…     

牙周炎龈下菌斑肽酶的椅旁快速检测

目的观察龈下菌斑肽酶活性与牙周临床指标的相关性，以探讨牙周炎活动性的监测方法。方法利用Periocheck椅旁快速检测60例成人牙周炎患者共72个位点龈下菌斑中牙龈卟啉菌、福塞氏类杆菌、齿垢密螺旋体来源的肽酶活性，与标准比色卡比较，区分阳性和阴性；在波长666nh下测吸光度值，定量反映肽酶活性。结果牙周炎位点肽酶活性(0.43±0.15)明显高于健康对照组(0.13±0.03)，差异有显著性(P＜0.01)；肽酶活性高低与探诊深度、附着丧失、牙龈指数呈正相关关系。结论Periocheck方法能够快速、简便、灵敏地检测龈下菌斑肽酶活性，反映牙周临床指标，可用于牙周炎活动性的监测。     

牙周炎患者龈下菌斑与口臭相关性的研究

目的　了解口臭与牙周炎患者龈下菌斑组成的关系。方法　用H2S测定仪Halimeter与专业医师鼻测两种方法对210例牙周炎患者进行口气测定,选取符合纳入标准的20例口臭患者作为实验组,另随机选取10例无口臭的牙周炎患者作为对照组。收集两组患者邻面间隙的颈缘菌斑和龈下菌斑行刚果红负性染色及厌氧培养,对培养出的主要细菌作分离及鉴定。所有实验结果采用SPSS 10.0进行统计学分析。结果　①两组患者的菌斑指数、牙龈出血指数、牙周袋深度无显著性差异。②两组患者邻面间隙的颈缘菌斑和龈下菌斑中螺旋体所占百分比均有显著性差异。③两组患者邻面间隙的颈缘菌斑及龈下菌斑中的细菌检出量均无显著性差异。④两组患者邻面间隙的颈缘菌斑及龈下菌斑中主要细菌的检出率均无显著差异;与口臭相关的细菌(牙龈卟啉单胞菌、韦荣菌和二氧化碳噬纤维菌)的构成比在邻面间隙的颈缘菌斑中为实验组显著高于对照组,而在龈下菌斑中两组无显著差异。结论　并非所有的牙周炎患者均有口臭;牙周炎患者邻面间隙的颈缘菌斑中细菌的构成比与口臭的关系更密切。     

悬突对牙周状态及龈下菌斑的影响

目的：从微生态学角度探讨悬突引起牙周炎的机制，揭示悬突对牙周组织的危害性。方法：检测568个有悬突牙及568个对照牙的G1和PD，采集龈下菌斑以刚果红染色，测其龋下菌群的细菌组成。结果：悬突牙组的GI、PD明显高于对照组，其螺旋体、杆菌及球菌较对照牙组有显著性差异。结论：悬突改变牙周微生态环境，使龋下菌群细菌组成发生变化；临床治疗中要加强悬突的防治。     

种植体周围龈下菌群的初步研究

作者将４８颗二段式圆柱形骨内种植体根据骨吸收程度分为甲组（０～２ｍｍ）乙组（〉２ｍｍ）另选健康天然牙为对照组（丙组）取龈沟内容物置于生理盐水内离心，用暗视野及Ｇｒａｍ染色镜检，对菌群进行了分析，研究，结果显示：健康种植体龈下菌群Ｇ球菌占８０％，而可动菌和螺旋菌极少，与对照组的天然牙相似。而不健康种植全组与对照组有较大的差异，以Ｇ杆菌占优势，多达６０％，螺旋体比例也显著增高。提示Ｇ＾－杆菌和螺旋体可     

乳牙菌斑附着部位的分析

目的观察分析全口乳牙的菌斑附着状况。方法应用菌斑染色片对120名5岁幼儿进行乳牙菌斑检查。结果120名儿童PI均值为1.69±0.01，男、女间无显著性差异；上后牙颊面PI最高；乳磨牙菌斑总量高于乳前牙；上颌牙唇颊侧菌斑附着多于腭侧、下颌牙舌侧多于唇颊侧。结论上述菌斑附着多的牙及牙面需强化刷牙。     

Comparison of plaque microflora between Chinese and Caucasian population groups

This investigation was designed to compare the predominant plaque micro-organisms from a Chinese group of patients exhibiting periodontitis with an age-, sex- and periodontal disease-matched Caucasian group of patients. In addition to race, the 2 population groups differed with respect to diet and oral hygiene habits, or effectiveness at removing plaque. Clinical measurements were determined along with an evaluation for micro-organisms in supragingival and subgingival plaque. Although the Chinese and Caucasian population groups were similar with respect to composition of micro-organisms in subgingival plaque, notable differences were observed in supragingival plaque. The Chinese group had higher mean proportions of spirochetes, motile rods. Fusobacterium spp. and dark-pigmented Bacteroides species, while the Caucasian group had higher mean proportions of cocci, total Actinomyces spp., A. viscosus and total Streptococcus spp. in supragingival plaque. The microbial differences observed in supragingival plaque may be explained at least in part, if not totally, by the higher plaque index scores of the Chinese versus Caucasian population groups.     

Ultrastructure of developing subgingival plaque in beagle dogs

The aim of this study was to describe the ultrastructure of developing subgingival plaque. In 6 beagle dogs, healthy gingiva prevailed after a pre-experimental period of intensive plaque control. At the start of the experiment, all oral hygiene measures were withdrawn. Biopsies comprising buccal gingiva and adjacent dental tissues were obtained from premolars and molars on days 0, 4, 7, 21, and 28 and processed for electron microscopy. Sections of day 0 exhibited a junctional epithelium in close contact with the tooth and absence of subgingival aggregations of bacteria on the tooth surface. For each of the subsequent periods studied, some sections did not exhibit subgingival plaque. In the remaining sections with subgingival plaque, almost all bacteria appeared in aggregates attached to the tooth surface. The number of bacteria at the orifice of the gingival sulcus increased with time. Further apically, the micro-organisms formed either a continuous layer of even thickness or, more frequently, discrete microcolonies along the tooth surface. The bacteria comprised practically entirely Gram-negative cocci, rods or spirochetes. The latter were present either mixed with the other types in the discrete microcolonies or constituted the bulk of the continuous layer of plaque. In the apical part of the subgingival plaque, a shift from Gram-negative cocci and rod during the early stages of gingival inflammation, to a predominance of spirochetes in later stages was noted. The study shows that in dogs, starting from a plaque- and gingivitis-free baseline, a predominantly Gram-negative subgingival flora may form within a few days after withdrawal of all oral hygiene measures and that a shift in the bacterial composition of the subgingival plaque takes place during a 4-week period of no oral hygiene.     

Combined antibiotic (metronidazole) and mechanical treatment effects on the subgingival bacterial flora of sites with recurrent periodontal disease

5 patients in maintenance, 1-3 years after periodontal therapy who showed sites with reinfected pockets and bleeding despite regular recall visits were selected. Darkfield microscopy from 3 sites in each patient showed an average of 41% spirochetes and 21% motile rods. Probing depths ranged from 7 to 9 mm and loss of clinical attachment from 6 to 13 mm in these sites. The patients were given 3 times 250 mg/day of metronidazole (Flagyl) for 10 days. Darkfield microscopy and microbiological cultures of the subgingival plaque were performed twice prior to the study, at the end of the medication and after 3 weeks, 3 and 6 months. The plaque and gingival indices, probing depth and loss of clinical attachment were recorded. During the medication and at 3 and 6 months, the teeth were scaled and root planed. The samples were obtained with 3 paper points and cultured anaerobically in the glove box on non-selective and selective media and representative bacterial colonies identified by aerobic growth, gram stain and rapid biochemical tests. Presumptive pathogenic micro-organisms including Bacteroides were identified. The % of spirochetes, motile rods and non-motile bacteria were enumerated by darkfield microscopy. The clinical results show that administration of metronidazole and repeated root planing significantly decreased gingival inflammation, probing depth and loss of clinical attachment in reinfected sites. After treatment, these sites harbored significantly less spirochetes and more non-motile bacteria, while motile rods tended to return to baseline levels with time. The combined antibiotic and mechanical therapy resulted in a statistically significant decrease of gram-negative rods, Fusobacteria and Bacteroides gingivalis over 6 months.(ABSTRACT TRUNCATED AT 250 WORDS)     

Use of monoclonal antibodies to enumerate spirochetes and identify Treponema denticola in dental plaque of children, adolescents and young adults

Identification of spirochetes in dental plaque is difficult. Not all spirochetes can be cultured and microscopic analysis based on darkfield or phase optics cannot determine the genus and species of individual bacterial cells. The purpose of this study was to use monoclonal antibodies in an immunoenzyme technique to stain spirochetes in dental plaque. Separate mAb were used to estimate total spirochetes and relative numbers of 2 distinct types of Treponema denticola. Plaque samples were collected from 40 subjects grouped by age. Results showed that older subjects are more likely to have spirochetes, to have more spirochetes and to have more diverse populations or spirochetes than younger subjects. Our studies suggest that T. denticola may be the first treponeme to colonize the primary dentition, that T. denticola appears to comprise a major proportion of all spirochetes at all ages and that two distinct serotypes of T. denticola are found to coexist in plaque from most children.     

Effect of metronidazole on development of subgingival plaque and experimental periodontitis

Active tissue destruction in experimental periodontitis has been positively correlated with subgingival spirochetes and total number of organisms. The present study was designed to inhibit spirochete populations and evaluate the effect upon periodontal destruction. Metronidazole was administered orally to four squirrel monkeys (100 mg/kg/bwt) for 17 days. After 3 days, marginal periodontitis was induced around bicuspids and molars by tying silk ligatures at the gingival margins. Subgingival plaque samples were taken baseline and 14 days after ligature placement. Dark-field microscopy quantitated motile forms, spirochetes, straight and curved rods, filament, cocci and fusiforms. Periodontal destruction was evaluated at 2 weeks by histometric analysis of connective tissue attachment, crestal alveolar bone and infiltrated connective tissue. Bacterial and histometric comparisons were made with experimental periodontitis data from four animals (control) which had not received metronidazole. Subgingival plaque prior to periodontitis induction was dominated by cocci, but fusiforms and straight rods were also present. Straight rods formed a greater proportion of the plaque which developed in metronidazole-receiving animals. The absence of spirochetes and motile rods contrasted with the control group where spirochetes were the predominant type. The total number of bacteria was also larger in the control animals. Histometric analysis showed that areas of infiltrated supracrestal connective tissue were similar in both groups. Loss of connective tissue attachment and alveolar bone was significantly less in experimental animals, and the latter values did not differ significantly from baseline dimensions. The results indicated that the subgingival bacterial populations which developed during metronidazole administration did not result in an experimental periodontitis.     

Microbial colonization patterns of loosely adherent subgingival plaque in adult periodontitis

Periodontal plaque is a complex bacterial ecosystem that carries an innate history of colonization, selection and maturation. Detailed examination of this balanced environment can reveal developmental sequences and certain interrelationships, much like an archeological record, that can provide insight in the understanding of plaque formation and maturation. For the present investigation, methods are employed which enable the retrospective elucidation of the historical data of plaque development and the nature of bacterial interactions. Nonparametric statistical methods are used to analyze risk, agreement and interdependence, following analytical techniques which are well established in medical epidemiology, but not generally employed in dentistry. The fundamental concept is that many organisms which are present in plaque prefer or require a pre-existing bacterial miliue for colonization and growth to steady-state level. Plaque samples and Ramfjord attachment level measurements were obtained from 60 adult periodontitis patients. Loosely adherent plaque was sampled and different morphotypes were enumerated by darkfield microscopy. The colonization of small spirochetes (S-SP) within the loosely adherent plaque was essential for the colonization of medium spirochetes (M-SP), odds ratio = 15.7 and filaments (FIL), odds ratio = 22.2. Thus, a temporal colonization sequence is inferred for FIL and M-SP, both requiring S-SP as a prerequisite morphotype. Medium spirochetes, in turn, are required for fusiform (FUS) colonization. M-SP also enhance the colonization of FIL and large motile rods (L-MO-R) within the loosely adherent plaque. These morphotypes were inferred to be sequentially interdependent, each preferring or requiring the presence of the preceding morphotype.(ABSTRACT TRUNCATED AT 250 WORDS)     

Darkfield microscopy of subgingival plaque of an urban black population with poor oral hygiene

A low socio-economic community residing in Crossroads, Cape Town, consists of people who originated from the Eastern Cape areas of the Ciskei and Transkei. These individuals have had virtually no dental care, with the exception of emergency treatment for pain. A darkfield microscopic study of a random sample of 52 individuals was undertaken to determine the predominant morphological forms in the subgingival plaque of this population. Spirochetes were found to constitute 42.1% and motile rods, 7.7%, of the total darkfield microscopic count. Clinical assessment of the periodontal status of 100 individuals revealed the following mean values; plaque index (PI) = 1.44, gingival index (GI) = 1.31, probing depth (PD) = 2.35 mm and loss of attachment = 0.64 mm. Subgingival calculus deposits were present in 42% of the group and little or no mobility of the teeth was evident. Poor oral hygiene resulted in 90% of the tooth surfaces of the sampled population in Crossroads being covered with plaque, yet these individuals appear to be resistant to periodontitis. The high proportions of spirochetes and motile rods found in the subgingival plaque of this group were not indicative of periodontitis and therefore, fail to confirm the findings of other investigators.     

The non-specific theory in microbial etiology of inflammatory periodontal diseases

In the absence of toothbrushing, the gingival crevice is colonized by a complex indigenous microflora causing gingivitis, a non-specific inflammation. Subgingival plaque may develop by downgrowth into the inflamed pocket of those micro-organisms from supragingival plaque which can multiply there. By direct microscopy, increased proportions of motile rods and spirochetes have been found in diseased pockets. Cultures on selective media have demonstrated increased prevalence of various gram-negative rods. Cultures on non-selective media have revealed the complexity and variability of the subgingival microflora, comprising more than 200 species. Destructive periodontitis is the result of subgingival colonization, which is favored by such ecological changes as plaque accumulation, gingivitis, and gingival exudate. These changes increase the numbers of micro-organisms and alter their proportions, but no single species appears in active sites which is not also commonly present in inactive sites. The subgingival micro-organisms have several virulence factors which promote colonization of the pockets, destroy host defense mechanisms, and provoke inflammation. It appears that different combinations of indigenous bacteria, rather than just a single species, can produce the pathogenic potential necessary to cause progression from gingivitis to destructive periodontitis.     

The effect of supragingival plaque control on the composition of the subgingival flora in periodontal pockets

The effect of mechanical supragingival plaque control on the composition of the subgingival microflora in untreated 4-6 mm deep periodontal pockets was investigated. 13 subjects with chronic periodontitis were recruited for the study. Periodontally-diseased sites were subjected to professional plaque control 3 x weekly for a period of 3 weeks. Contralateral sites received no prophylaxis and served as controls. No instructions in oral hygiene procedures were given to the patients who maintained their habitual oral hygiene regime during the observation period. Clinical examination and darkfield microscopic analysis of bacterial samples were performed every week. The PlI scores for the experimental sites were reduced markedly, while those for the control sites remained stable throughout the observation period. No changes in the other clinical parameters were detected during the study. The composition of the subgingival microflora at the control sites did not change during the experimental period. In contrast, at the test sites, the proportion of spirochetes+motile rods decreased continuously. This decrease reached statistical significance at the end of the experiment. The results indicate that at periodontally diseased sites with an established subgingival ecosystem, supragingival plaque removal may influence the composition of the subgingival microflora.     

The microbial morphotypes associated with periodontitis health and adult periodontitis: composition and distribution

Subgingival plaque samples were obtained from 162 sites in 27 adult periodontitis patients and 162 sites in 27 healthy patients using a standardized lavage technique. The distribution of 10 different microbial morphotypes was determined by darkfield microscopy. The lavage technique selectively samples the loosely adherent plaque at the base of the periodontal pocket and not the tooth-associated, adherent plaque. This standardized technique permits quantitative comparisons of numerical density of morphotype composition at different sites, in addition to qualitative comparisons or relative proportions. There was a significant positive association between the numerical density of each morphotype within the non-adherent plaque and the number of sites at which the organism was detected in both healthy and diseased subjects. A previously undescribed darkfield morphotype, has been detected with this method. This morphotype, a small motile coccobacillus (S-MO-CB) has been found to be the numerically dominant species in both health and disease. This morphotype has been recovered in pure culture following passage through a 0.4 mu filter and includes organisms of the Wollinella and Campylobacter genus. Non-motile organisms comprised less than 1-2% of the sample from healthy and diseased sites. Motile forms, such as spirochetes, had a high frequency of detection in healthy individuals. Analysis of pooled plaque samples revealed that the prevalence of cocci and fusiforms was significantly elevated in patients with healthy periodontium, as compared to patients with adult periodontitis. In adult periodontitis patients, the frequency of occurrence of medium spirochetes, filaments and small nonmotile rods was significantly elevated in pooled plaque. Analysis of individual sites indicated that the proportion and numerical density of most morphotypes within the non-adherent plaque were not significantly different in disease as compared to health. Disease is characterized by an increased % of small spirochetes and fusiforms at each site. At diseased sites which harbor small spirochetes, the numerical density is elevated four-fold, as compared to healthy sites which have small spirochetes. The numerical density of other morphotypes is not significantly different comparing healthy sites to diseased sites. Thus, the increase in the % of small spirochetes in disease in due to a site-localized four-fold increase in numerical density within the non-adherent plaque.