牙周致病菌的共聚及其对人工牙根面黏附力的影响

目的比较具核梭杆菌、牙龈卟啉单胞菌、中间普雷沃菌和伴放线嗜血菌等牙周病致病菌彼此之间的共聚力大小,观察四者在具核梭杆菌介导下对人工牙根面黏附力的影响,了解牙周生物膜结构中细菌间可能存在的相互作用。方法目测具核梭杆菌、牙龈卟啉单胞菌、中间普雷沃菌和伴放线嗜血菌彼此间的共聚力,以放射性核素闪烁计数四者在具核梭杆菌黏附和未黏附状况下对胶原包被羟磷灰石（c—HA）的黏附间是否存在着差异。结果具核梭杆菌、牙龈啉单胞菌、中间普雷沃菌和伴放线嗜血菌彼此间存在着共聚作用,其中,具核梭杆菌与牙龈卟啉单胞菌、牙龈卟啉单胞菌与中间普雷沃菌间的共聚度均可达4度,具核梭杆菌与其他三菌间的共聚度均大于3度。牙龈卟啉单胞菌、中间普雷沃菌和伴放线嗜血菌在具核梭杆菌黏附的状况下对c—HA的黏附率高于其在具核梭杆菌未黏附时的黏附率,具核梭杆菌在未黏附的状况下对c—HA的黏附率高于其在黏附后的黏附率。结论具核梭杆菌、牙龈卟啉单胞菌、中间普雷沃菌和伴放线嗜血菌彼此间均存在共聚关系,具核梭杆菌可能对其他牙周病致病菌定植于牙菌斑起到了桥梁作用。     

牙周可疑致病菌对胶原包被羟磷灰石黏附的体外实验研究

目的比较对牙根面具有黏附能力的具核梭杆菌、牙龈卟啉单胞菌、中间普氏菌和伴放线菌嗜血菌对胶原包被的羟磷灰石实验膜（C- HA）的黏附能力,初步探讨以上牙周可疑致病菌在牙根表面形成菌斑生物膜的能力。方法采用同位素闪烁计数法测定上述4种细菌黏附至C- HA表面的黏附量及黏附率,比较其黏附能力。结果培养相同时间,不论是培养24 h还是培养48 h,4种不同细菌两两比较,具核梭杆菌ATCC 10953和牙龈卟啉单胞菌ATCC 33277对C- HA表面黏附率的差异无统计学意义;中间普氏菌ATCC 25611和伴放线菌嗜血菌ATCC 29523的黏附率之间差异也无统计学意义,但是具核梭杆菌ATCC 10953和牙龈卟啉单胞菌ATCC 33277对C- HA表面的黏附率显著高于中间普氏菌ATCC 25611和伴放线菌嗜血菌ATCC 29523（P<0.001）。同一种细菌,在培养不同时间即培养24 h和48 h,对C- HA表面黏附率的差异均无统计学意义。结论不同的牙周可疑致病菌对胶原包被的羟磷灰石的选择性黏附作用不同,具核梭杆菌和牙龈卟啉单胞菌对胶原有较强的亲和作用,在细菌的局部定植过程和牙周炎的进展和复发中可能发挥重要作用。     

抗牙龈卟啉单胞菌IgY的制备及抑菌效果

目的:使用牙龈卟啉单胞菌(P.g)诱导产蛋母鸡特异性IgY抗体产生及制备,特异性IgY抗体抑制P.g及其它牙周病病因菌生长.方法:采用免疫接种、水稀释、盐析、液体培养抑菌及ELISA等方法,诱导、提纯IgY抗体,抑制P.g及其它牙周病病因菌生长.结果:诱导产生的IgY抗体经硫酸铵盐析提取纯度达87.6%～89.1%,IgY特异性结合牙龈卟啉单胞菌抗原的结合效价1∶ 1 600.制备的抗牙龈卟啉单胞菌IgY抗体与牙龈二氧化碳噬纤维菌、中间普氏菌、伴放线放线杆菌、具核梭杆菌等牙周病致病菌的交叉免疫反应抗原结合效价分别为1∶ 800、1∶ 800、1∶ 6 400、1∶ 12 800.抗牙龈卟啉单胞菌IgY在5.0、1.0、0.1 g/L时,分别与牙龈卟啉单胞菌、牙龈二氧化碳噬纤维菌、伴放线放线杆菌、中间普氏菌、具核梭杆菌、粘性放线菌、变形链球菌等厌氧菌在(1×108) CFU/L和(5×108) CFU/L时培养24 h和72 h均有不同程度的抑制其生长作用.结论:牙龈卟啉单胞菌免疫产蛋母鸡诱导产生的特异性IgY抗体在一定的浓度内有抑制牙龈卟啉单胞菌生长,以及抑制多种牙周病致病菌生长的作用.牙龈卟啉单胞菌与这些牙周病病因均存在着共同抗原,其可能具有防治牙周病的前景.     

次氯酸钠溶液抗菌体积分数的测定

目的评价次氯酸钠溶液对牙龈卟啉单胞菌、中间普雷沃菌、具核梭杆菌和粪肠球菌牙髓病原菌的体外抗菌性能。方法采用琼脂稀释法测定次氯酸钠溶液对牙龈卟啉单胞菌、中间普雷沃菌、具核梭杆菌和粪肠球菌的抗菌活性,其抗菌活性以最低抑菌体积分数(MIVF)和最低杀菌体积分数(MBVF)表示。结果次氯酸钠溶液对牙龈卟啉单胞菌、中间普雷沃菌、具核梭杆菌和粪肠球菌的MIVF分别为0.035%、0.07%、0.018%和0.28%,MBVF分别为0.035%、0.07%、0.035%和0.28%,其总的抗菌效果排序分别为具核梭杆菌、牙龈卟啉单胞菌、中间普雷沃菌和粪肠球菌。结论次氯酸钠对口腔常见厌氧菌有抑制和杀灭作用,具有理想的抗菌性能。0.5%MBVF的次氯酸钠溶液的抑菌性能可满足临床需求。     

安止痛牙龈膏对牙龈卟啉单胞菌和伴放线嗜血菌及具核梭杆菌的抑菌作用

目的 体外测定安止痛牙龈膏对牙龈卟啉单胞菌、伴放线嗜血菌和具核梭杆菌的抑制活性.方法 以微量液体稀释法,测定安止痛牙龈膏对上述3种致病菌的最小抑菌质量浓度(MIC)和最小杀菌质量浓度(MBC).结果 安止痛牙龈膏对上述3种致病菌均有抑制作用.其中,对伴放线嗜血菌的抑制作用最强,MIC和MBC均为0.8g·L-1;对牙龈...     

黄芩对5种常见牙周细菌抑制作用的体外研究

目的：观察黄芩对5种常见牙周细菌的作用。方法：采用试管两倍稀释法测定黄芩甲醇提取物在体外厌氧环境对牙龈卟啉单胞菌、中间普氏菌、伴放线放线杆菌、具核梭杆菌、血链球菌的最小抑菌浓度(MIC)和最小杀菌浓度(MBC)。结果：黄芩对各实验菌株均有抑制作用，对牙周常见可疑致病菌牙龈卟啉单胞菌、中间普氏菌、伴放线放线杆菌、具核梭杆菌的MIC值为5g／L、MBC值为20g／L，对牙周有益菌血链球菌的MIC值为40g/L、MBC值为40g/L，结论：浓度为5g/L的黄芩在不破坏牙周局部生态平衡的情况下可有效抑制牙周可疑致病菌的生长。     

牙周炎的厌氧菌群的分离及对抗生素的敏感性研究

目的：调查牙周炎思考口腔厌氧菌群的分布并研究其对β内酰胺药物的敏感性。方法：选择44例牙周炎患者，对其龈下菌斑中的厌氧菌进行分离培养，用7种β内酰胺药物对分离到的厌氧菌进行敏感性试验。结果：与牙周病有关的厌氧菌分离率为100％，产黑色素普雷沃菌86．4％，中间普雷沃菌56．8％，具核梭杆菌63．6％，放线菌38．6％，微小消化链球菌52．3％，牙龈卟啉单胞菌65．9％，二氧化碳嗜纤维菌9．1％。厌氧菌对各类β内酰胺药物保持敏感。结论：产黑色素普雷沃菌、中间普雷沃菌、具核梭杆菌、微小消化链球菌、牙龈卟啉单胞菌和放线菌，是牙周炎思考牙龈沟中的优势菌；β内酰胺药物可作为厌氧菌牙周炎的治疗药物。     

壳聚糖共混物对4种牙周主要致病菌体外抑菌实验

目的：评价不同质量比的壳聚糖和葡甘聚糖共混物对4种牙周常见致病菌的抑菌作用。方法通过纸片法测定不同质量比壳聚糖/葡甘聚糖共混物对4种牙周炎致病菌牙龈卟啉单胞菌、中间普氏菌、伴放线放线杆菌和具核梭杆菌标准菌株的体外抑制实验,探讨壳聚糖/葡甘聚糖共混物对上述4种细菌的抑制作用。结果不同质量比的壳聚糖/葡甘聚糖共混物对4种牙周致病菌均有抑菌作用,但作用效果不同,随壳聚糖质量比的下降,抑菌效果也随之下降。4种不同牙周炎致病菌对壳聚糖/葡甘聚糖共混物敏感性有差异,伴放线放线杆菌敏感性最高,中间普氏菌次之,牙龈卟啉单胞菌再次之,具核梭杆菌最差。结论不同质量比的壳聚糖/葡甘聚糖共混物可不同程度抑制上述4种牙周致病菌生长。     

五倍子对5种常见牙周细菌抑制作用的体外研究

目的：观察五倍子对5种常见牙周细菌的作用。方法：采用试管二倍稀释法，测定五倍子水提取物在体外厌氧环境对牙龈卟啉单胞菌，中间普氏菌，伴放线放线杆菌，具核俊杆菌和血链球菌的最小抑菌浓度（MIC），结果：五倍子对各实验细菌均有抑制作用。对牙周常见可疑致病菌牙龈卟啉单胞菌，伴放线放线杆菌，中间普错菌，具核梭杆菌的MIC值均为3．12％，对牙周有益菌血链球菌的MIC值则为12．5％，结论：浓度为3．12％的五倍子在不破坏牙周局部生态平衡的情况下可有效抑制牙周细菌的生长。     

牙周致病菌密度感应信号系统luxS基因的检测

目的 检测牙周可疑致病菌密度感应信号系统luxS基因,了解其在牙周致病菌中的分布.方法 选取牙龈卟啉单胞菌、伴放线放线杆菌、具核梭杆菌的模式株、参考株及临床分离株作为研究对象,提取DNA,通过聚合酶链反应(PCR)、电泳鉴定和DNA测序,并利用GenBank数据库的Blast检测以上细菌luxS基因的存在情况.结果 电泳鉴定存在目的 条带,测序和Blast检测表明牙龈卟啉单胞菌PCR产物与目的 基因有高度一致性(均为99%以上),具核梭杆菌测序结果 与GenBank数据库的基因相同,伴放线放线杆菌电泳鉴定结果 显示存在目的 条带(750 bp),与参考条带大小一致.结论 本实验引物设计合理,能较好地扩增出牙龈卟啉单胞菌、具核梭杆菌、伴放线放线杆菌各实验菌株的luxS基因,为进一步研究luxS基因的功能奠定了基础.     

二甲胺四环素胶原制剂体外抗菌研究

目的研究胶原蛋白为载体复合二甲胺四环素制成的牙周缓释制剂的抗菌效果,以期寻找辅助治疗牙周炎的新制剂。方法采用杯碟法对含20mg/g盐酸二甲胺四环素的胶原牙周缓释剂和派丽奥软膏进行牙周常见致病菌的抑菌实验,测定最小抑菌浓度（MIC）和抑菌环直径。菌珠选用国际标准菌株牙龈卟啉单胞菌、中间型普里沃氏菌、巨核梭形杆菌、伴放线放线杆菌和粘性放线菌。结果胶原牙周缓释制剂和派丽奥软膏对各实验菌株均有抑菌作用,牙龈卟啉单胞菌、中间型普里沃氏菌、巨核梭形杆菌、伴放线放线杆菌的MIC为0.31mg/L,粘性放线菌的MIC为1.2mg/L。2种药物比较抑菌效果无显著性差异（P〉0.05）。结论胶原牙周缓释制剂对牙周主要致病菌具有良好的抑制作用。     

Antimicrobial profiles of periodontal pathogens isolated from periodontitis patients in The Netherlands and Spain

BACKGROUND AND AIM: Antimicrobial resistance of periodontal pathogens towards currently used antibiotics in periodontics has been investigated in a previous study. Microbial resistance in the periodontal microflora was more frequently observed in Spanish patients in comparison with Dutch patients. The aim of the present study was to compare antimicrobial susceptibility profiles of five periodontal bacteria isolated from periodontitis patients in Spain and in The Netherlands. MATERIAL AND METHODS: Subgingival plaque samples from adult patients with periodontitis were collected and cultured on selective and non-selective plates. Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, Prevotella intermedia, Fusobacterium nucleatum and Micromonas micros were isolated and used for minimal inhibitory concentration tests using the Epsilometer (E-test) technique. Eight different antibiotics were tested on all bacterial isolates. MIC50 and MIC90 values for each antibiotic and each species were determined and the percentage of resistant strains was calculated. RESULTS: Significantly higher MIC values were noted in Spanish strains of F. nucleatum for penicillin, ciprofloxacin, of P. intermedia for penicillin, amoxicillin and tetracycline, of M. micros for tetracycline, amoxicillin and azithromycin, and of P. gingivalis for tetracycline and ciprofloxacin. Based on breakpoint concentrations, a higher number of resistant strains in Spain were found in F. nucleatum for penicillin, amoxicillin and metronidazole, in Prevotella intermedia for tetracycline and amoxicillin, and in A. actinomycetemcomitans for amoxicillin and azithromycin. Resistance of P. gingivalis strains was not observed for any of the antibiotics tested both in Spain and The Netherlands. CONCLUSIONS: Differences exist in the susceptibility profiles of periodontal pathogens isolated from periodontitis patients in Spain and in The Netherlands. This implicates that antibiotic susceptibility testing is necessary to determine efficacy of antimicrobial agents. Also, clinical studies with antibiotics should take these differences into account. The information from the present study indicates that it may not be possible to develop uniform protocols for usage of antibiotics in the treatment of severe periodontitis in the European Union.     

Viability of four putative periodontal pathogens and enteric rods in the anaerobic transport medium VMGA III

The aim of this study was to determine the survival in VMGA III of Porphyromonas gingivalis, Prevotella intermedia, Fusobacterium nucleatum, Actinobacillus actinomycetemcomitans and enteric rods in laboratory cultures as well as in sub-gingival plaque samples. Laboratory strains of the 4 putative periodontal pathogens and Escherichia coli were used in the laboratory part of the study. Also, 31 subgingival plaque samples were obtained from 22 periodontal patients and stored in VMGA III. Each sample, from both the laboratory and the clinical parts, was divided into 3 portions. One portion was cultured within a few hours of collection (baseline), while the second was processed after 24 h (day 2) and the third 48 h later (day 3). The results of the clinical part indicate that the detection frequencies of all 4 periodontal pathogens and their levels in positive samples decreased, to different degrees, by day 2 and decreased further by day 3. Enteric rods were not detected in baseline samples. However, they were present in 16.1% and 22.6% of day 2 and day 3 samples, respectively. Similarly, the laboratory results demonstrate a significant decrease in the levels of the 4 periodontal pathogens tested by day 2 and day 3, whereas the opposite occurred for E. coli. P. gingivalis, P. intermedia , and F. nucleatum survived better in the presence of E. coli than alone, whereas A. actinomycetemcomitans survived less well when co-inoculated with E. coli . VMGA III appears to maintain microbial population ratios for periods up to 24 h. After 24 h, the multiplication of enteric organisms may alter the original proportions of the sample.     

Aggregatibacter actinomycetemcomitans serotype f O-polysaccharide mediates coaggregation with Fusobacterium nucleatum

BACKGROUND/AIMS: Intergeneric bacterial coaggregation may play an important role in plaque development. METHODS: In this study we investigated the coaggregation reaction between two periodontal pathogens, Aggregatibacter actinomycetemcomitans and Fusobacterium nucleatum. RESULTS: Previous studies showed that A. actinomycetemcomitans serotype b strains coaggregate with F. nucleatum strain PK1594, and that A. actinomycetemcomitans serotype b O-polysaccharide (O-PS) is the receptor responsible for coaggregation between A. actinomycetemcomitans and F. nucleatum. A. actinomycetemcomitans serotype f O-PS has been shown to be structurally and antigenically related to serotype b O-PS. In the present study we show that A. actinomycetemcomitans strain CU1060N, a serotype f strain, also coaggregated with F. nucleatum PK1594. Like coaggregation between serotype b strains and F. nucleatum, coaggregation between CU1060N and F. nucleatum was inhibited by galactose. An O-PS mutant of CU1060N failed to coaggregate with F. nucleatum. CONCLUSION: We concluded that A. actinomycetemcomitans serotype f O-PS, like serotype b O-PS, mediates coaggregation between A. actinomycetemcomitans and fusobacteria.     

Attachment of Fusobacterium nucleatum PK1594 to mammalian cells and its coaggregation with periodontopathogenic bacteria are mediated by the same galactose-binding adhesin

It has been shown that Fusobacterium nucleatum PK1594 coaggregates with Prophyromonas gingivalis PK1924 through a galactose-binding adhesin. In the present study, attachment of F. nucleatum PK1594 to a variety of mammalian cells was characterized. F. nucleatum PK1594 attached to all eukaryotic cells tested, including human buccal epithelial cells, gingival and periodontal ligament fibroblasts, HeLa cells and murine lymphocytes, macrophages, and polymorphonuclear leukocytes. These attachments were (i) inhibited by galactose, lactose and N-acetylgalactosamine and (ii) inhibited by monoclonal antibody specific for the galactose-binding adhesin of F. nucleatum PK1594. In addition, a coaggregation-defective mutant of F. nucleatum PK1594 (PK2172), which does not exhibit galactose binding activity, did not attach to the mammalian cells. Coaggregation of F. nucleatum PK1594 with P. gingivalis PK 1924 and Actinobacillus actinomycetemcomitans JP2, but not with other bacteria, showed a similar pattern with sugars, monoclonal antibody, and the adhesin-deficient mutant. The results suggest that the attachment of F. nucleatum PK1594 to mammalian cells and its coaggregation with periodontal pathogens are mediated by the same galactose-binding adhesin.     

Multicenter evaluation of tetracycline fiber therapy. III. Microbiological response

In a multicenter study of the effects of tetracycline (TC) fiber therapy, subgingival plaque samples were tested for 6 probable periodontal pathogens by DNA probe analysis. Levels of Actinobacillus actinomycetemcomitans, Eikenella corrodens, Fusobacterium nucleatum, Porphyromonas (Bacteroides) gingivalis, Prevotella intermedia (Bacteroides intermedius), and Wolinella recta were quantitatively determined in samples taken at baseline, and immediately after TC fiber removal, control fiber removal, and scaling and root planing. At untreated sites, samples were taken at baseline and 10 d later. Specificity of the DNA probe method was evaluated by testing the hybridization to 83 reference cultures. Interaction of the F. nucleatum probe with Fusobacterium periodonticum, and of the W. recta probe with Wolinella curva were the only cross-hybridizations noted. Species were detected at an average sensitivity of 2.9 x 10(4) organisms per sample. Approximately 70% of sites were initially infected with P. gingivalis and F. nucleatum; 50% with P. intermedia and E. corrodens; infections with W. recta and A. actinomycetemcomitans were less common (36% and 11% respectively). The average numbers of organisms found in the plaque samples were highest for F. nucleatum, P. gingivalis, and P. intermedia (ca. 10(6)). E. corrodens, W. recta, and A. actinomycetemcomitans occurred at 10-fold lower levels. Bacterial numbers and proportions of species in subgingival sites from the five centers did not differ appreciably. Both TC fiber therapy and scaling decreased the number of sites infected with all the monitored species. The bacterial composition at untreated sites and at sites where control fibers were placed was not significantly altered. The percentage reduction of the number of sites with detectable infection varied with each species: from 86% with W. recta to approximately 40% with P. gingivalis. Significant reduction of pocket depth and bleeding occurred at TC fiber-treated sites infected with each of the species. Significant attachment level gain occurred only at sites initially infected with P. gingivalis and treated with TC fibers.     

新型盐酸米诺环素缓释凝胶对牙周致病菌的缓释抑菌作用

目的:评价新型盐酸米诺环素缓释凝胶对牙周主要致病菌的体外缓释抑菌效果。方法:采用超声乳化法制备20 g/L盐酸米诺环素缓释凝胶及非缓释凝胶,以琼脂纸片扩散法检测二者在1、3、5 h和1、3、5、7 d七个时间点的释放液对Pg、Fn、Av、Pi.的抑菌活性,测量抑菌环直径。结果:缓释凝胶组对于Pg、Fn、Av、Pi抑菌作用均可持续到7 d;而非缓释凝胶组对Pg、Fn、Av的抑菌作用仅能维持到3 d,对Pi的抑菌作用可持续到5 d。结论:新型盐酸米诺环素纳米缓释凝胶对Pg、Fn、Av、Pi抑菌作用均可持续到7 d以上,具有较好的缓释抑菌效果。     

Prevalence of periodontal pathogens with varying metabolic control of diabetes mellitus

Abstract. The purpose of this study was to determine the prevalence of 5 periodontal pathogens in individuals with diabetes mellitus. Subjects ( n = 107) 20–70 years of age with type 1 ( n = 60) or 2 ( n = 47) diabetes mellitus were studied for the occurrence of the periodontal pathogens A. actinomycetemcomitans, F. nude-alum, E. corrodens, P. gingivalis and P. intermedia. Subgingival plaque was sampled in each subject from a single site exhibiting the greatest inflammation. The evaluation of selected periodontal bacterial pathogens was based on an immunoassay utilizing bacterial specific monoclonal antibodies. 35% of the sites harbored P. gingivalis , 28% F. nucleatum and 21% E. corrodens. A. actinomycetemcomitans and P. intermedia were found in less than 10% of the sites. Subjects for whom the probing depth at the sampled site was 4 mm were more often found to have detectable pathogens than those with a probing depth 3 mm. Diabetic factors such as duration, type and metabolic control of the disease had no statistically significant effect on the prevalence of these bacteria.