Streptococcal diversity in oral biofilms with respect to salivary function

OBJECTIVE: In a previous study, we screened 149 subjects and established four groups high or low for salivary killing of oral bacteria, and for aggregation and live and dead adherence of oral bacteria (as a combined factor). Caries scores were significantly lower in both High Aggregation-Adherence groups. In this study we looked at the effects of those differences in salivary function on the quantity and diversity of oral biofilm streptococci. DESIGN: Subjects from those four groups were recalled for collection of overnight oral biofilm from buccal upper central incisors, lingual lower central incisors, buccal upper and lower first molars, and lingual upper and lower first molars. At each site, groups were compared for total biofilm (by DNA concentration), total streptococci (by quantitative PCR), and streptococcal diversity (by Streptococcus-specific denaturing gradient gel electrophoresis). RESULTS: Total biofilm DNA and total streptococci were correlated. Both were highest on buccal molar surfaces and lowest on lingual lower central incisors, and both were significantly lower in the High Aggregation-Adherence groups (particularly at the buccal molar site). Fifty distinct bands were observed in denaturing gradient gels. There was great diversity within and between sites. Three major bands were present in almost every person at every site. Densities for two of those bands were significantly lower in both High Aggregation-Adherence groups. Other less-prevalent bands also showed the same pattern. CONCLUSION: These findings are consistent with our caries results in suggesting that differences in salivary function can influence the quantity and composition of streptococci in oral biofilms.     

Distribution and prevalence of mutans streptococci in the human dentition

The prevalence of mutans streptococci was determined in 14,859 samples of plaque from all available tooth surfaces in 114 subjects. The clinical examination included location of incipient caries lesions, fillings, and crowns. Mutans streptococci were detected on 40% of all tooth surfaces. The frequency distribution of mutans streptococci and the level of colonization showed a decreasing gradient from molars to incisors for buccal, lingual, occlusal, and approximal surfaces. The location and number of approximal restorations were closely related to the colonization level of mutans streptococci except for second and third molars. Restored surfaces tended to be more colonized by mutans streptococci than sound surfaces, except for occlusal surfaces. A high prevalence of mutans streptococci was found in plaque samples from tooth-colored fillings, especially on buccal and lingual surfaces.     

Streptococci dominate the diverse flora within buccal cells

Previously, we reported that intracellular Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, and Tannerella forsythensis were present within buccal epithelial cells from human subjects, as lesser components of a polymicrobial flora. In this study, we further characterized that intracellular flora by using the same double-labeling techniques to identify Fusobacterium nucleatum, Prevotella intermedia, oral Campylobacter species, Eikenella corrodens, Treponema denticola, Gemella haemolysans, Granulicatella adiacens, and total streptococci within buccal epithelial cells. All those species were found within buccal cells. In every case, species recognized by green-labeled species-specific probes were accompanied by other bacteria recognized only by a red-labeled universal probe. Streptococci appeared to be a major component of the polymicrobial intracellular flora, being present at a level from one to two logs greater than the next most common species (G. adiacens). This is similar to what is observed in oral biofilms, where diverse species interact in complex communities that often are dominated by streptococci.     

Protective properties of salivary pellicles from two different intraoral sites on enamel erosion

The purpose of this study was to investigate the protective effect and ultrastructure of salivary pellicles formed in vivo near the orifices of the ducts of parotid and submandibular/sublingual salivary glands. Pellicles were formed by exposing bovine enamel slabs to the oral environment at the buccal aspect of the upper first molars and at the lingual aspect of the lower incisors in 3 subjects over periods of 24 h. Enamel specimens with and without 24-hour pellicles were immersed in citric acid (0.1 and 1%) for periods ranging from 30 s to 5 min, and processed for measurement of surface microhardness (SMH) and transmission electron microscopy (TEM). In comparison to uncovered enamel specimen significantly less decrease in SMH due to acid exposure was observed in pellicle-coated enamel specimens. Pellicles formed at the buccal aspect of the upper molars were less effective in protecting the enamel against acid-induced softening as compared to pellicles formed at the lingual aspect of the lower incisors only after 5 min exposure in 1% citric acid. TEM analysis showed that pellicle layers were dissolved continuously due to acid exposure. However, even after 5 min exposure to 1% citric acid, a residual pellicle layer could be detected on the enamel surface. In conclusion, site-dependent differences of buccally and lingually in vivo formed 24-hour pellicles have minor importance concerning the pellicle-induced protection of the enamel surface against erosive changes.     

Localized aggressive periodontitis in primary dentition: a case report

BACKGROUND: A 5-year-old Japanese boy presented with persistent gingival inflammation and severe mobility of the right lower primary incisors. Due to severe alveolar bone loss and a deep periodontal pocket (5 mm), the incisors were extracted at the second visit. METHODS: Clinical, radiographic, histological, and microbiological examinations were carried out. Then, the polymerase chain reaction (PCR) technique was employed to detect specific periodontal pathogens. The chemotactic activity of polymorphonuclear neutrophils was also measured. RESULTS: Tannerella, Capnocytophaga, Fusobacterium, and Eikenella sp. were recovered from the subgingival microflora around the right lower incisors, while A. actinomycetemcomitans, Tannerella forsythensis (formerly Bacteroides forsythus), Prevotella nigrescens, Campylobacter rectus, and Capnocytophaga gingivalis were detected using the PCR method. Further chemotaxis assay revealed that neutrophil function was depressed compared with that of healthy controls. CONCLUSIONS: Although inflammation remained around the right primary second molars, the bone loss was controlled by periodic professional mechanical teeth cleaning (PMTC), subgingival irrigation, and local antibiotic application. The probing depths of all teeth, including permanent incisors and molars, were within 2.5 mm.     

Occurrence of Porphyromonas gingivalis and Tannerella forsythensis in periodontally diseased and healthy subjects

BACKGROUND: The purpose of this study was to compare the prevalence and level of Porphyromonas gingivalis (P. gingivalis) and Tannerella forsythensis (T. forsythensis) in subgingival plaque samples from both healthy individuals and periodontal patients in different age groups. METHODS: A total of 498 subgingival plaque samples were studied. These samples were collected from 407 individuals diagnosed with periodontal disease (210 adult periodontitis [AP], 78 rapidly progressive periodontitis [RPP], and 119 refractory periodontitis [Ref-P] cases) and 91 healthy (H) subjects. P. gingivalis and T. forsythensis were detected by indirect immunofluorescent assay using species-specific polyclonal antisera to P. gingivalis strain (FDC 381) and T. forsythensis strain (FDC 335). The prevalence of P. gingivalis and T. forsythensis was compared by chi-square analysis. Differences in P. gingivalis and T. forsythensis levels among various periodontal status and age groups was determined by one-way analysis of variance and Fisher's multiple comparison tests. The association between the presence of P. gingivalis or T. forsythensis in different periodontal status and age groups was measured using odds ratios. RESULTS: P. gingivalis was found in 85.7% (P < 0.0001) and T. forsythensis in 60.7% (P = 0.0002) of diseased subjects compared to 23.1% and 39.6%, respectively, in healthy subjects. P. gingivalis, but not T. forsythensis, was detected more frequently in any diseased group than in the H group in every age group (P<0.0001). No significant difference was found in the prevalence of P. gingivalis and T. forsythensis among age groups, except T. forsythensis was more prevalent in the age group of 40 to 59 years than in the age group < 20 years (chi2 = 3.93, P = 0.047) in the H group. The mean level of P. gingivalis and T. forsythensis was significantly higher in diseased groups than in the H group (P < 0.0001). The odds ratio for P. gingivalis in the AP group (25.0) was greater than any other group for P. gingivalis or T. forsythensis compared to the H group. CONCLUSIONS: These data suggested that P. gingivalis is closely associated with the pathogenesis of periodontitis and that it may not be a normal inhabitant of periodontally healthy subjects. T. forsythensis is also important in the pathogenesis of periodontal disease; however, whether it causes periodontal disease or is a secondary invader of periodontal lesions remains to be determined.     

Identification of periodontopathic bacteria in gingival tissue of Japanese periodontitis patients

INTRODUCTION: The identification of invading periodontopathic bacteria in tissues is important to determine their role in the pathogenesis of periodontal disease. The objective of this study was to identify periodontopathic bacteria in diseased gingival tissue of periodontitis patients. METHODS: Subgingival plaque and gingival tissue were collected from 32 generalized chronic periodontitis (CP), 16 generalized aggressive periodontitis (GAgP) and eight localized aggressive periodontitis (LAgP) patients. Detection frequencies and quantities of Porphyromonas gingivalis, Actinobacillus actinomycetemcomitans and Tannerella forsythensis were investigated by polymerase chain reaction. The prevalences of Streptococcus oralis and Streptococcus sobrinus were also examined and the distribution of A. actinomycetemcomitans serotypes was observed. RESULTS: P. gingivalis and T. forsythensis were detected in approximately 70% of tissue samples and 50% of plaque samples in the three periodontitis groups. Prevalence of A. actinomycetemcomitans in tissue samples was higher in the LAgP (63%) group than in either the CP (16%) or the GAgP (38%) group. A. actinomycetemcomitans serotype c was detected in 50% of LAgP patients. Detection frequencies of S. oralis and S. sobrinus were markedly low in both plaque and tissue samples from all three periodontitis groups. Amounts of P. gingivalis, A. actinomycetemcomitans and T. forsythensis in the tissue samples were not different among the three periodontitis groups. CONCLUSION: P. gingivalis, A. actinomycetemcomitans and T. forsythensis can localize in diseased gingival tissue and may be involved in periodontal tissue destruction. Serotype c is the predominant serotype of A. actinomycetemcomitans in Japanese LAgP patients.     

Regional differences in the detection rate of periodontopathic bacteria in supragingival plaque

Although there are numerous reports on detection of bacteria causing periodontal diseases within the oral cavity, most of these treat the oral cavity as a single entity. The objective of this study was to determine whether there was site-specificity in the detection of certain bacteria in supragingival plaque from different regions of the oral cavity. The subjects were six adults with no recognized oral problems. We examined eight regions; the labial and lingual surfaces of the upper and lower anterior teeth and the upper and lower first molars. The target bacteria included five species of bacteria that cause periodontal diseases. After professional medical tooth cleaning (PMTC), the volunteers performed no oral prophylaxis for three days. Then we collected about 1 μl of plaque from each of the regions by excavators. We searched for the five species of bacteria by immunoslot-blot assay and compared the results from the respective regions. We discovered that the detection rate was very small for all five bacterial species on the lingual surface of the lower anterior teeth, which is known to have good salivary clearance. On the labial surface of the upper anterior teeth, which is commonly believed to have poor salivary clearance, the detection rate was higher but there was no specificity with respect to detection rate. The regions with the highest detection rates were the buccal surface of the upper and lower molars. On the lower molars, larger populations of Porphyromonas gingivalis and Prevotella nigrescens were detected. On the upper molars, Prevotella intermedia and Prevotella melaninogenica were detected. These results suggest that there is regional specificity in the presence of periodontopathic bacteria in the mouth.     

慢性牙周炎龈下菌斑中五种牙周可疑致病微生物的分布

目的研究五种牙周可疑致病微生物在慢性牙周炎患者龈下菌斑的分布。方法选择27例慢性牙周炎患者,每位患者选取牙周袋最深的两个位点作为观察位点,采集龈下微生物样本,采用多重聚合酶链反应和反杂交的方法对伴放线菌嗜血菌、牙龈卟啉单胞菌、福赛斯坦纳菌、中间普雷沃菌和齿垢密螺旋体五种微生物进行半定量检测。结果在所检测的54个位点中,牙龈卟啉单胞菌、中间普雷沃菌、福赛斯坦纳菌和齿垢密螺旋体均有较高的检出率,分别为98.15%、92.59%、100%和98.15%;伴放线菌嗜血菌检出率较低,为20.37%。牙龈卟啉单胞菌和福赛斯坦纳菌的检出量明显高于其他三种微生物,其差异有统计学意义（P<0.05）。结论慢性牙周炎患者多存在牙龈卟啉单胞菌、福赛斯坦纳菌、中间普雷沃菌和齿垢密螺旋体的同时感染,且牙龈卟啉单胞菌和福赛斯坦纳菌的感染量较高。     

Quantitative real-time polymerase chain reaction versus culture: a comparison between two methods for the detection and quantification of Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis and Tannerella forsythensis in subgingival plaque samples

OBJECTIVE: The purpose of this investigation was to validate a real-time quantitative polymerase chain reaction (PCR) assay in identifying and quantifying Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis and Tannerella forsythensis from subgingival plaque samples taken from subjects with different periodontal conditions, when compared with conventional cultural procedures. PATIENTS AND METHODS: Ninety-two adult subjects participated in this study, 32 with periodontitis, 30 with gingivitis and 30 healthy. A pooled subgingival sample was obtained from every patient. Culturing procedures were carried out using standard techniques. For real-time PCR analysis, primers were selected from sequences of the LktC (A. actinomycetemcomitans), Arg-gingipain (P. gingivalis) and BspA antigen (T. forsythensis) genes. Contingency tables were constructed to compare the qualitative results, while quantitative data were evaluated by paired t-test. RESULTS: A. actinomycetemcomitans was the least frequently recovered species with both techniques. Prevalence of P. gingivalis was low in healthy patients, increased in gingivitis and peaked in periodontitis patients. The frequency of detection of T. forsythensis showed marked differences between culture and PCR, although the same tendency of an increase in prevalence from health to gingivitis and to periodontitis was observed with both methods. Contingency tables demonstrated a good level of agreement between PCR and culture procedures for A. actinomycetemcomitans and P. gingivalis, especially in periodontitis patients. P. gingivalis culture counts were significantly higher than those obtained by PCR. The opposite was true for T. forsythensis, and statistically significant higher counts were obtained by PCR for gingivitis and periodontitis patients. CONCLUSION: This study demonstrated a good agreement between the quantitative PCR technology and the culture procedure. The high sensitivity and specificity of the quantitative PCR technology justify its use in epidemiological studies and as an adjunct in clinical diagnosis of periodontal patients.     

Relationship between mutans streptococci in saliva and their colonization of the tooth surfaces

The relationship between the salivary concentration of mutans streptococci and their prevalence on different tooth surfaces was studied in 114 subjects. Plaque samples were obtained from all tooth surfaces in the dentition and the infection magnitude of mutans streptococci was determined. The salivary concentrations of mutans streptococci correlated significantly with the number of colonized tooth surfaces and with the infection level of mutans streptococci for individual teeth or groups of tooth surfaces. The highest correlation values were found for buccal and approximal surfaces and for molars followed by premolars and anterior teeth. The 10 tooth surfaces best reflecting the salivary levels of mutans streptococci were 5 buccal and 5 approximal sites, 6 of them localized on maxillary posterior teeth. A significant positive relationship was noted between the prevalence of mutans streptococci in saliva and on the dorsum of the tongue.     

Microscopic analysis of the effects of ultrasonic instrumentation on the root surface of molar furcations

Twenty molars, 10 from the maxilla and 10 from the mandible with furcation areas type II (Staffileno) were instrumented in their inter-radicular area with P10 instrument of Cavitron. It was established that the furcations of easier access instrumentation were the lingual o lower teeth and the buccal of the upper, and the most difficult accessibility were the distal of upper molars. The instrumentation can leave grooves or deformations, depending on the instrument to the area. It was concluded that ultrasonic instrument are useful removing supragingival calculus and bacterial plaque, and it must be limited to perform such procedures.     

Incidence of selected ureolytic bacteria in human dental plaque from sites with differing salivary access

Saliva is the main source of urea in the human mouth and may be responsible for the predilection of ureolytic bacteria for certain tooth sites. As a test of this hypothesis, the ureolytic bacteria, Haemophilus parainfluenzae, Actinomyces naeslundii, Actinomyces viscosus and coagulase-negative oral staphylococci, were enumerated in supragingival plaque from various sites in each of 10 subjects. The sites sampled included the maxillary and mandibular incisors (chosen because the lower incisors are more exposed to the submandibular-sublingual secretion than the upper) and the maxillary and mandibular molars (the upper molars being closer to the source of parotid saliva). After dispersion of the plaque samples in saline, subsamples of each suspension were plated on appropriate selective media and other subsamples were taken for nitrogen analysis to measure the amount of plaque sampled. H. parainfluenzae that used urea was present in the largest numbers, A. viscosus was next and A. naeslundii and coagulase-negative staphylococci were least. The staphylococci and H. parainfluenzae were more numerous from mandibular than from maxillary incisors and from maxillary than mandibular molars, a pattern which suggests that salivary access favours their selection. The numbers of A. viscosus and A. naeslundii were not related to salivary access: A. viscosus was most numerous from the maxillary incisors, possibly because this site is normally the most acidic of the four studied and A. viscosus is strongly acidogenic and aciduric; the incidence of A. naeslundii had no relationship with site.(ABSTRACT TRUNCATED AT 250 WORDS)     

New cavity forms in first deciduous molars which maximize dentin thickness between cavity and pulp chamber

Using a total of 60 human extracted first deciduous molars (30 upper molars and 30 lower molars), we contrived a variety of ideal cavities having dentin thicknesses below the cavity (subcavitary dentin) that measure approximately 1 mm in thickness from the pulp chamber at any point of measurement and also having a retentional groove prepared in such a way that detachment of a filling material is prevented. The transparent specimens prepared from the cavity-containing teeth were cut into serial sections of 93 microns. These sections were reconstructed using a personal computer. The thickness of dentin below the cavity was measured in randomly selected sections. Results obtained were as follows: 1. An ideal thickness of the subcavitary dentin was preserved for upper first deciduous molar by preparing a retentional groove lingually or by providing a dovetail-like shape to mesio-distolingual sides in the cavity; for lower first deciduous molar, any form of cavitation worked. 2. In the upper first deciduous molars, the margins of the cavity were displaced medially to the summits of the respective cusps 1.9 mm at the buccal side and 1.2 mm at the lingual side. In the lower first deciduous molars, the buccal margin medially measured 0.9 mm and the lingual margin measured 0.6 mm at the mesial side. At the distal side, the buccal and lingual margins measured 1.2 mm and 1.0 mm, respectively. 3. The ratio of cavity width to the distance between the summits of the buccal and lingual cusps was 1/3 in upper deciduous first molars and 2/5 in the lower first deciduous molars. 4. In the upper first deciduous molars, the depths of the buccal and lingual walls of the cavity at the center of the central groove were both 1.1 mm. In the lower first deciduous molars, the cavity formed with its center at the middle of the transverse ridge had a depth of 1.5 mm at the buccal wall and a depth of 1.7 mm at the lingual wall. The cavity formed with its center at the central fossa had a depth of 1.2 mm at the buccal wall and 1.1 mm at the lingual wall. 5. The width of the gingival wall in the proximal box measured 0.6-0.7 mm in upper and lower first deciduous molars.     

Characteristics of periodontal microflora in acute myocardial infarction

BACKGROUND: Periodontitis has been linked to increased risk of cardiovascular diseases. Systemic reactions associated with cardiovascular events may depend on characteristics of the subgingival microflora in periodontitis. Our objectives were to compare the numbers of cultivable bacteria, composition of subgingival microflora and clonal distribution of Actinobacillus actinomycetemcomitans (A. actinomycetemcomitans) in two groups of patients with generalized chronic periodontitis (GCP), one with an acute myocardial infarction (AMI-GCP) and the other one without AMI (non-AMI-GCP). METHODS: In all, 150 dentate individuals were screened for suitability to this study. Subgingival bacterial samples were collected from 11 AMI-GCP and 11 non-AMI-GCP patients who had been selected using strict inclusion criteria in an attempt to exclude confounding factors and to increase comparability of periodontal conditions by matching for periodontal probing depths and attachment levels. Culture methods were used to determine the total viable counts and occurrence and proportions of six periodontal bacterial species and yeasts. Polymerase chain reaction (PCR) technique was used to detect A. actinomycetemcomitans and Porphyromonas gingivalis (P. gingivalis). Intraspecies characterization of A. actinomycetemcomitans included serotyping and genotyping. RESULTS: The mean proportions of P. gingivalis (P = 0.05) and Tannerella forsythensis (T. forsythensis) (P = 0.01) were significantly lower, but the numbers of Micromonas micros (M. micros) and A. actinomycetemcomitans were up to nine times higher and the mean total number of cultivable bacteria per sample higher (P <0.01) in AMI-GCP than in non-AMI-GCP. CONCLUSION: The findings that no target subgingival species were overrepresented but the total bacterial number was higher in AMI-GCP than non-AMI-GCP patients may provide support to the hypothesis that elevated numbers of bacteria in close vicinity to sterile parenteral area present a risk for systemic health.     

舌侧矫治器对牙周致病菌的影响

目的研究舌侧矫治器对患者牙周临床指标和牙周致病菌的影响。方法收集成年正畸治疗患者55例资料,28例使用颊侧矫治器作为对照组,27例使用舌侧矫治器作为试验组,于治疗前和治疗6个月后,分别记录菌斑指数、龈沟出血指数、探诊深度,PCR检测龈下菌斑中牙龈卟啉单胞菌(Porphyromonas gingivalis,Pg)、伴放线放线杆菌(Actinobacillus actinomycetemcomtans,Aa)、福赛斯坦氏菌(Tannerella forsythensis,Tf)3种牙周致病菌的检出率。结果治疗6个月试验组菌斑指数、龈沟出血指数、探诊深度分别为2.36±0.71、2.05±0.49、(3.43±0.56)mm,对照组分别为1.86±0.44、1.67±0.25、(2.87±0.74)mm,2组间差异均有统计学意义(P<0.05);试验组Pg、Aa检出率分别为37.0%和22.2%,对照组的Pg、Aa检出率分别为14.3%和10.7%,试验组高于对照组(P<0.05)。结论舌侧矫治器,较颊侧矫治器,对牙周临床指标影响更大,可造成更多的牙周致病菌聚集。     

The detection of Porphyromonas gingivalis, Prevotella intermedia and Actinobacillus actinomycetemcomitans in tooth, tongue and buccal mucosa plaques in children, using immunoslot blot assay (IBA)

The present study was to investigate the distribution of typical periodontpathic bacteria (Porphyromonas gingivalis, Prevotella intermedia and Actinobacillus actinomycetemcomitans) in tooth, tongue and buccal mucosa plaques in 3 to 17 Year old children. Clinical parameters (Rates of df, d, DMF, and D; plaque and gingival index) for each subject were determined prior to the collection of each site plaque. Three periodontopathic bacteria on each site samples were detected using IBA. The frequency of three bacteria for tooth plaque was higher than that for tongue or buccal mucosa plaque. The frequency of Porphyromonas gingivalis and Prevotella intermedia in supragingival plaques was significantly higher than that of corresponding ones in tongue or buccal mucosa plaques. The three bacteria also occurred more frequently in subjects aged between 10 and 14 years. Periodontopathic bacteria may be enhanced in circumpubertal children.     

长春市某小学7～12岁儿童牙周致病菌分布状态调查

目的 应用聚合酶链反应(PCR)法对儿童口腔内牙龈卟啉单胞菌(Porphyromonas gingivalis,Pg)、伴放线放线杆菌(Actinobacillus actinomycetemcomitans,Aa)分布状态进行检测,探讨检出结果与牙周临床指标之间的关系.方法 选取长春市自强小学151名7至12岁儿童为研究对象,选择右上颌中切牙唇面和右上颌第一磨牙颊面为被检部位,取龈上菌斑、记录探诊出血(bleeding on probing,BOP)、探诊深度(probing depth,PD)、牙龈指数(gingival index,GI),应用PCR法对两菌种进行检测.结果 ①儿童龈上菌斑中Pg、Aa检出率为27.6%、54.3%;②6颊面Pg、Aa的检出率(40.0%、57.9%)均高于1 唇面(15.5%、50.7%),Pg检出率差异有统计学意义(P<0.01),且与BOP、PD、GI呈正相关;③Pg检出率随年龄增长呈逐渐增高趋势,Aa检出率在11～12岁组最高,其次为7～8岁组和9～10岁组;④BOP阳性部位Pg、Aa检出率(38.3%、65.4%)均高于BOP阴性部位(23.2%、50.5%),P<0.05.在BOP阳性部位,随PD加深Pg检出率逐渐增高,特别是在PD≥4mm时,Pg检出率明显增高(P<0.05),显示Pg检出率与BOP阳性、PD增加呈正相关.结论 7～12岁儿童龈上菌斑中高频度分布着Pg、Aa;上颌前牙区与磨牙区菌丛构成不同,Pg在磨牙区定植更早;两菌种检出率随年龄增长而增加,且与牙周临床指标密切相关,儿童早期采取牙周病的预防措施是非常必要的.     

Comparison of culture and real-time PCR for detection and quantification of five putative periodontopathogenic bacteria in subgingival plaque samples

OBJECTIVES: Bacterial cultivation is a well-established method for analyzing plaque samples. Real-time polymerase chain reaction (PCR) is a novel rapid method for the identification and quantification of periodontopathogenic bacteria that has been recently introduced. In this study, we compared real-time PCR with conventional anaerobic cultivation. METHOD: A total of 78 subgingival plaque samples were harvested from pockets > or =5 mm in 22 patients with advanced chronic periodontitis and immediately transferred into transport medium. Aliquots were evaluated with species-specific probes by real-time PCR (meridol Perio Diagnostics, GABA) and anaerobic bacteria culture on selective media for the detection of Actinobacillus actinomycetemcomitans, Fusobacterium nucleatum, Porphyromonas gingivalis, Prevotella intermedia and Tannerella forsythensis. The analysis was performed by two separate, blinded examiners. RESULTS: When real-time PCR was compared with the culture method (golden standard) for the detection of putative periodontopathogenic bacteria, the sensitivity and specificity for A. actinomycetemcomitans were 67% and 100%, respectively (kappa: 0.79); for F. nucleatum 73% and 53%, respectively (kappa: 0.21); for P. gingivalis 94% and 84%, respectively (kappa: 0.77); for P. intermedia 33% and 94%, respectively (kappa: 0.26) and for T. forsythensis 92% and 56%, respectively (kappa: 0.51). Spearman's correlation coefficients for the quantitative results of both methods were 0.82 for A. actinomycetemcomitans, 0.33 for F. nucleatum, 0.83 for P. gingivalis, 0.38 for P. intermedia and 0.67 for T. forsythensis. CONCLUSION: Overall, the agreement between both test methods was excellent for A. actinomycetemcomitans and P. gingivalis, fair for T. forsythensis and poor for F. nucleatum and P. intermedia. The discrepancies in the results may be explained by the inability of cultivation methods to distinguish between close related taxa, and the problems of keeping periopathogenic bacteria viable, which is required for standard cultivation.     

Scanning electron microscopy evaluation of aligner fit on teeth

Objectives:The fitting of aligners on anchorage teeth is a crucial factor in clear aligner orthodontics. The purpose of this experimental study was to evaluate the fitting of two aligner systems, Invisalign and CA-Clear Aligner, using scanning electron microscopy (SEM).Materials and Methods:Passive aligners (Invisalign and CA-Clear Aligner) were adapted on resin casts obtained by stereolithography (STL) files of a patient, and then sectioned buccolingually. Upper and lower central incisors, upper and lower first premolars, and upper and lower first molars were the regions analyzed. Representative microphotographs of sections were taken with a scanning electron microscope (SEM); a total of 160 micrometric measurements were obtained and analyzed with ANOVA tests.Results:Invisalign provided an overall better fit on lower incisors (F = 11.48, P = .0095) and on lower molars (F = 19.93, P = .0012). Considering the different regions, Invisalign provided better fit at the gingival edge of the buccal aspect on lower incisors (F = 11.33, P = 0.0056) and at the gingival edge of the lingual aspect on upper premolars (F =5.34, P = 0.0047). On the upper molars, Invisalign provided better fit at the gingival edge of the buccal aspect, while CA-Clear Aligner provided better fit at the buccal maximum convexity, on the buccal cusp, on the occlusal groove and at the palatal cusp. On lower molars, Invisalign showed a more accurate fit at the buccal aspect points.Conclusions:Invisalign and CA-Clear Aligner exhibited comparable fit on anchorage teeth. Invisalign provided better fit at the gingival edges of aligners, while the CA-Clear Aligner provided better fit on complex occlusal surfaces.