Salivary IgA response to probiotic bacteria and mutans streptococci after the use of chewing gum containing Lactobacillus reuteri.

We investigated whether ingestion of probiotic bacteria could influence salivary IgA levels, specific anti-mutans streptococci IgA levels and specific antibodies towards the ingested probiotic bacterium. The study was a randomised, double-blind, placebo-controlled trial, where the test group (n = 11) received twice daily chewing of gum containing Lactobacillus reuteri (2 × 10(8)  CFU per dose) and the control group (n = 12) received placebo. Resting saliva was collected before and after 12 weeks of treatment and 4 weeks after end of treatment. Total salivary IgA concentrations were measured by ELISA. Specific IgA reactivity was determined using a whole-cell ELISA. Results were expressed as % IgA per protein in saliva. The level of total IgA% per protein increased significantly between pretreatment levels (13.5%) and follow-up treatment levels (14.4%) within the test group only (P < 0.05). No changes were seen in the control group during the trial. The level of probiotic-reactive antibodies decreased significantly between pre- and post-treatment samples (from 12.2% to 9.0%, P < 0.05) in the test group. Similarly, the level of specific mutans streptococci antibodies decreased significantly between pre- and post-treatment samples (P < 0.05) in the test group only (for Streptococcus mutans from 20.1% to 15.0%; for Streptococcus sobrinus from 7.4% to 5.3%). Ingestion of probiotic bacteria might influence the adaptive immune response of the host.

Alpha-1-antitrypsin deficiency and periodontitis, a pilot study.

The aim of this study was to investigate if periodontal parameters and elastase in gingival crevicular fluid (GCF) are different in alpha-1-antitrypsin deficient (AATD) subjects compared to subjects with normal AAT level. Thirty subjects were included, 20 of whom with severe AATD, phenotype PiZZ. Ten AATD subjects suffered from chronic obstructive pulmonary disease (COPD, group 1) and 10 were asymptomatic (group 2). Ten control subjects, phenotype PiMM, (group 3) were recruited from a public dental clinic. The examination comprised of sampling of GCF, Gingival Index (GI), Plaque Index (PlI), probing pocket depth (PPD) and radiography. GCF was collected with paper strips (Periopaper). Plasma AAT concentration was measured by nephelometry and AAT in GCF with ELISA. Elastase activity and protein in GCF were determined by spectrophotometry. The mean values for GI, PlI, PPD and the radiological measurements did not show any statistically significant differences between the groups. AAT in plasma and GCF demonstrated very low values in groups 1 and 2 with no significant difference between these groups but a statistical difference in comparison with group 3. Elastase in GCF did not show any difference between the three groups. In conclusion, neither the periodontal parameters nor the elastase in GCF were different in AATD subjects, phenotype PiZZ, when compared to subjects with normal AAT level, phenotype PiMM, in this material.

Influence of dietary supplementation with Lactobacillus reuteri on the oral flora of healthy subjects.

UNLABELLED: Investigate the presence of Lactobacillus reuteri in saliva after supplementation with L. reuteri and the probiotic effect of L. reuteri on plaque index and supra- and subgingival microbiota. MATERIAL AND METHODS: The study included 23 healthy individuals, randomised into test or control subjects. At baseline and after 12 weeks saliva samples, plaque index and supra- and subgingival plaque samples were obtained. The test subjects were given the study product (containing L. reuteri, ATCC 55730 and ATCC PTA 5289) and the control subjects placebo for 12 weeks. Microbiological analyses were done by checkerboard DNA-DNA hybridization technique and selective culturing for lactobacilli determination. RESULTS: A significant increase in total Lactobacillus counts in saliva occurred in both groups (p < 0.05) with a significant increase of L. reuteri (p = 0.008) in the test group.Termination of intervention resulted in a wash out of L. reuteri. The control group demonstrated a statistically significant increase in PII after 12 weeks (p = 0.023) whilst there was no significant change in the test group. A significant increase was found for most bacterial species in both groups in supra- and subgingival plaque with no significant difference for any of the species between the groups. The ratio between "bad/good" supragingival bacteria decreased for the test group but this decrease did not reach significance. The corresponding ratio for subgingival bacteria decreased significantly in both groups. Supplementation of L. reuteri resulted in presence of L. reuteri in saliva but L. reuteri was washed out after termination of intervention. No significant effect on supra- or subgingival microbiota was observed. The significant increase in PII in the control group with no significant change in the test group may, however, indicate a probiotic effect of L. reuteri in this study population.

Effects of ovariectomy and aging on tooth attachment in female mice assessed by morphometric analysis.

OBJECTIVE: Non-human primates, dogs, rats, hamsters and ferrets, have frequently been used as laboratory animals in periodontal biology and pathology. In the past, mice have been used less for this purpose, but nowadays attract a lot of interest because gene knockout and transgenic technologies utilize mice primarily. In this study, we investigate the effects of ovariectomy and aging on tooth attachment in female mice. MATERIAL AND METHODS: Eight-week-old mice (n=15) were divided into three experimental groups (control, n=5; sham-operated, n=5; ovariectomy, n=5) and ovaries removed bilaterally. Attachment level, assessed by measuring alveolar bone height and apical termination of the junctional epithelium, was determined 6 weeks post-ovariectomy by digital morphometric analysis in sagittal sections of the mandible. The plasma level of the inflammation marker serum amyloid A (SAA) was determined by ELISA. In another series of experiments, tooth attachment was determined in female mice (n=7) at 8-26 weeks of age. RESULTS: Withdrawal of female sex hormone production by ovariectomy had no effect on alveolar bone height and apical termination of the junctional epithelium. The SAA level in plasma was unaffected by removal of the ovaries, suggesting that systemic inflammation is not induced by ovariectomy. Bone height was similar in mice sacrificed at 8-26 weeks of age and apical termination of the junctional epithelium was at the cemento-enamel junction at all ages. CONCLUSIONS: Removal of ovarian production of female sex hormones by ovariectomy has no influence on tooth attachment, and further tooth attachment is preserved with age in female mice.

LPS-induced MCP-1 and IL-6 production is not reversed by oestrogen in human periodontal ligament cells.

OBJECTIVE: Periodontal ligament (PDL) cells express oestrogen receptors but the functional importance of oestrogen in PDL cells exposed to bacterial endotoxins is not known. Here we investigate if the inflammation promoter lipopolysaccharide (LPS) affects PDL cell production of interleukin-6 (IL-6), monocyte chemoattractant protein-1 (MCP-1), C-reactive protein (CRP) and/or normal functional PDL cell characteristics such as collagen synthesis and cell proliferation and if oestrogen modulates the effects of LPS. METHODS: PDL cells were obtained from periodontal ligament of premolars. PDL cells were treated with Escherichia coli LPS in the absence or presence of oestrogen (17beta-oestradiol, E2). Cellular concentration of IL-6, MCP-1 and CRP was determined by enzyme-linked immunosorbent assay (ELISA). Collagen synthesis was determined by l-[3H]proline incorporation. Cell proliferation was assessed by DNA synthesis measurement using [3H]thymidine incorporation. RESULTS: Stimulation with LPS (500 ng/ml to 10 microg/ml) increased IL-6 production in a concentration-dependent manner. Lower concentration (100 ng/ml) of LPS had no effect. LPS-induced stimulation of IL-6 was not reversed by a physiologically high concentration (100 nM) of E2. LPS increased also MCP-1 production which was unaffected by E2. Treatment with E2 alone had no effect on either IL-6 or MCP-1. Stimulation with LPS had no effect on CRP. LPS did not affect collagen synthesis and cell proliferation, reflecting normal physiological properties of PDL cells. CONCLUSIONS: LPS stimulates PDL cell IL-6 and MCP-1 production but has no effect on the normal physiological properties of PDL cells. LPS-induced IL-6 and MCP-1 is not reversed by oestrogen suggesting that oestrogen exerts no anti-inflammatory effect via this mechanism.

Demonstration of mitochondrial oestrogen receptor beta and oestrogen-induced attenuation of cytochrome c oxidase subunit I expression in human periodontal ligament cells.

OBJECTIVE: Periodontal ligament (PDL) cells express oestrogen receptor beta (ERbeta) protein, but cellular functions regulated by ERbeta in these cells have not been identified. In this study we determine if ERbeta is localised to mitochondria and if oestrogen regulates mitochondrial function in human PDL cells obtained from teeth extracted for orthodontic reasons. DESIGN: Subcellular distribution of ERbeta was determined by confocal microscopy of cells co-stained with ERbeta antibody and the mitochondrion-selective probe MitoTracker and by immunogold electron microscopy. Expression of the mitochondrial enzyme cytochrome c oxidase subunit I, involved in oxidative phosphorylation, was determined by Western blotting in cells treated with or without physiological concentrations of the endogenous oestrogen 17beta-oestradiol. RESULTS: ERbeta immunoreactivity was observed both in the nuclei and the cytoplasm. MitoTracker-labelling was observed in the cytoplasm, especially in the perinuclear region, but not in the nuclei. Co-localisation of ERbeta and MitoTracker was observed in cells derived from both male and female subjects. Mitochondrial localisation of ERbeta was confirmed by immunogold electron microscopy. Cells treated with or without 17beta-oestradiol (100 nM) displayed an identical pattern of staining for mitochondria. Treatment with 100 nM 17beta-oestradiol attenuated cytochrome c oxidase subunit I expression by about 30%, while combined treatment with 17beta-oestradiol and the ER blocker ICI 182780 (10 microM) had no effect. CONCLUSION: This study demonstrates mitochondrial localisation of ERbeta and oestrogen-induced decrease in the expression of cytochrome c oxidase subunit I in human PDL cells, suggesting that oestrogen probably via ERbeta influences mitochondrial function and PDL cell energy metabolism.

Analysis of the interleukin-1 and interleukin-6 polymorphisms in patients with chronic periodontitis. A pilot study.

The aim of this study was to analyse whether the interleukin-1 (IL-1) and IL-6 gene polymorphisms were associated with the susceptibility of chronic periodontitis. Genomic DNA was obtained from 20 patients with chronic periodontitis and 31 periodontally healthy subjects. All subjects were of North European heritage. The test subjects were kept in a maintenance program after periodontal treatment but yet showing signs of recurrent disease. Genotyping of the IL-1alpha [+4845C>T], IL-1beta [-3954C>T] and IL-6 [-174G>C] polymorphisms was carried out using an allelic discrimination Assay-by-Design method on ABI PRISM 7900 Sequence Detection System. All genotypes were analyzed using the GeneMapper 2.0 software. A similar distribution of Single Nucleotide Polymorphism (SNP) was seen in both groups. Analysis by logistic regression including gender, IL-1alpha [+4845C>T], IL-1beta [-3954C>T], IL-6 [-174G>C] genotypes, the composite IL-1 genotype, the combination of the composite IL-1 genotype and the IL-6 -174G>C genotype and adjusting for smoking did not result in any statistically significant difference. SNPs in IL-1alpha [+4845C>T], IL-1beta [-3954C>T] and IL-6 [-174G>C] do not seem to increase the susceptibility to chronic periodontitis in this group of subjects.

The effect of Swedish and American smokeless tobacco extract on periodontal ligament fibroblasts in vitro.

Use of moist snuff is widespread in Sweden. In 2004 approximately 8oo,ooo Swedes were daily users which corresponds to 22% of the male population and 3% of the female population. The aim of the present study was to evaluate the effect of Swedish moist snuff extract on PDLfibroblast growth and hard tissue production and compare with moist snuff extract from USA. Periodontal ligament cells (PDL-cells) were obtained from 3 healthy subjects (1 female 14 years, 2 males 14 and 17 years) from the root surface of premolars extracted for orthodontic reasons. The cells were isolated from explants and grown in Dulbecco's Modified Eagle's Medium (DMEM) supplemented with 10% fetal calf serum (FBS) and cultivated in 37 degrees C with 5% CO2 in air. Snuff extract in concentrations 0.3%, 1% and 3% (in DMEM with 1% FBS) was tested. Cells from each individual were tested three times, each time in triplicate. Photographs were taken at o and 24 hours with a digital camera and analysed in terms of growth and morphology. Then the cell suspension was frozen and later thawed for examination of the production of alkaline phosphatase after exposure to different snuff concentrations. This in vitro study has shown that PDL cells from 3 different subjects demonstrated a reduced number of cells at exposure to 3% of both Swedish and American snuff extract. The production of alkaline phosphatase after 2 hours was similarly reduced from cells exposed to 3% snuff extract. Further studies have to be made to understand the effect of smokeless tobacco on periodontal tissues. However, from this study can be concluded that smokeless tobacco has biological effects in terms of reduced PDL cell growth and production of alkaline phosphatase

Clinical consequences of IL-1 genotype on early implant failures in patients under periodontal maintenance.

BACKGROUND: Implant failure and biologic complications such as periimplantitis are not completely avoidable. Are there any genetic and microbiologic parameters that could be used to identify patients at risk for implant failure, preferably prior to treatment? This would result in improvement of the diagnostics, treatment decision, and risk assessment. PURPOSE: The aims of this retrospective study were to describe (1) the absolute failure rate of Brånemark System implants (Nobel Biocare AB, Göteborg, Sweden) consecutively installed over a 10-year period in partially edentulous patients treated for periodontal disease prior to implant treatment and under regular professional maintenance, (2) the rate of interleukin-1 (IL-1) polymorphism in those patients who experienced at least one implant failure during the first year of function, and (3) the prevalence of periodontal pathogens in dental and periimplant sites with and without signs of inflammation. MATERIAL AND METHODS: Of 766 patients, 81 encountered at least one implant failure; 22 patients were clinically examined and were tested genetically for IL-1 genotypes. The presence of Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, and Prevotella nigrescens was analyzed. RESULTS: The absolute implant survival rate for the whole population was 95.32%; 10.57% of the patients encountered an implant loss. Implant loss in the examined group (n = 22) was 32 of 106 (30.1%); 10 (45%) of the 22 patients were smokers, and 6 (27%) of the 22 patients were IL-1 genotype positive. Patients positive for IL-1 genotype were not more prone to implant loss; however, a significant synergistic effect with smoking was demonstrated. Between patients who were IL-1 genotype positive and those who were IL-1 genotype negative, the differences in regard to bleeding on probing or periodontal pathogens did not reach statistical significance. CONCLUSION: The overall implant failure rate in a population treated and maintained for periodontal disease is similar to that of healthy subjects. A synergistic effect found between smoking and a positive IL-1 genotype resulted in a significantly higher implant loss. This indicates that further research with a larger patient group should focus on multifactorial analysis for adequate risk assessment.

The microbial outcome observed with polymerase chain reaction in subjects with recurrent periodontal disease following local treatment with 25% metronidazole gel.

The aim of this study was to evaluate the microbial outcome in patients with recurrent periodontal disease following treatment with 25% metronidazole gel using the polymerase chain reaction (PCR). Twenty subjects in a maintenance care program but with recurrent periodontal disease participated. Three months after scaling and root planing a total of 40 sites, 2 in each patient, with pocket probing depth of > or = 5 mm were selected. One site randomly selected was treated with 25% metronidazole gel (test) and the other site with a placebo gel (control). A bacterial sample was collected on paperpoint from each test and control site at baseline and 12 weeks after treatment. The following pathogens were analysed and detected with PCR:Actinobacillus actinomycetemcomitans (A.a.), Porphyromonas gingivalis (P.g.) and Prevotella nigrescens (P.n.). At baseline, A.a., P.g. and P.n. were detected in 30, 60 and 70% of all test sites and in 32, 58 and 21% of all control sites. There was a statistically significant difference between the test and control sites for P.n. at baseline. The major difference after treatment with 25% metronidazole gel was the increase of positive control sites for P.g. and P.n. However, there were no statistically significant differences in the occurrence rate of A.a., P.g. and P.n. at test and control sites after treatment. This study has shown that 25% metronidazole gel treatment did not seem to influence the microbial outcome, when PCR was used to analyse the presence/absence of A.a., P.g. and P.n. in this group of subjects with recurrent periodontal disease.

Implication of cervical enamel projection to furcation involvement in molars. A pilot clinical study.

The aim of this study was to investigate the correlation of cervical enamel projection (CEP) with furcation involvement (FI) and compare the healing response of molars with or without CEP after surgery. A total of 30 patients contributing 78 maxillary and mandibular first or second molars were included. Plaque Index (PII), Gingival Index (GI), probing pocket depth (PPD) and probing attachment level (PAL) were measured before surgery and 1 and 3 or 6 months postoperatively. During surgery, CEPs were identified and classified with a modified grading system from Masters & Hoskins (24). FI was measured horizontally from the buccal aspect into the furcation with a graduated probe to the nearest mm. Any measurement > or = 1 mm was considered as FI. CEPs were found in 33 molars (42%). Grade III CEPs were found in 14 teeth, Grade IIIb in 4 teeth, Grade II in 1 tooth and Grade I in 14 teeth. The results showed no significant correlation of CEP with FI. Nor was CEP significantly affecting the PPD and PAL 3 or 6 months after surgery. However, FI was a significant factor in the further loss of PAL after surgery. Further studies, involving larger sample size may be necessary in order to give more conclusive results.

Prevalence of cervical enamel projection and its correlation with furcation involvement in eskimos dry skulls.

The objectives of this study were to investigate the prevalence of cervical enamel projection (CEP) in molars of Eskimo dry skulls and to study the correlation of CEP with furcation involvement (FI). The material consisted of 834 upper and lower first and second permanent molars from 133 Eskimo dry skulls. CEPs were investigated from the buccal aspect of the tooth and classified according to a system modified from Masters & Hoskins (12). FI was measured horizontally from the buccal aspect into the furcation with a graduated probe to the nearest mm. Any measurement > or = 2 mm was considered to have positive FI. The result showed a presence of 72% of CEPs among the examined molars. Grade III was found in 53%, Grade II in 9% and Grade I in 11% of the 834 molars. Lower molars had a higher prevalence of CEPs (78%) than upper molars (67%). With the individual skull used as the unit for analysis, a statistically significant correlation of CEP with FI was found in upper right 2nd molar, upper left 1st molar, lower left 1st and 2nd molars and lower right 1st molar. These results may be of clinical importance since the impact of CEPs to periodontal treatment of FIs has been discussed.

Pontes removíveis (construçäo em cones) em pacientes com periodontile severa: relato de caso / Removable bridges (bulding as cones) in patients with severe periodontitis: report of a case

No presente trabalho, os autores descrevem a técnica de Pontes Removíveis (construçäo em cones), relatando um caso clínico de periodonite severa, em que foi empregada esta técnica após a terapia periodontal. Dividem o tratamento em duas partes (clínica e de laboratório), descrevendo as diversas fases técnicas adotadas e indicam o material instrumental necessário, referindo-se a necessidade de utilizaçäo de articuladores semi-ajustáveis, paralelômetro e outros pertences de registro