Neutrophils in chronic and aggressive periodontitis in interaction with Porphyromonas gingivalis and Aggregatibacter actinomycetemcomitans.

BACKGROUND AND OBJECTIVE: This study analyzed the interaction of Porphyromonas gingivalis ATCC 33277 and Aggregatibacter actinomycetemcomitans Y4 with peripheral blood polymorphonuclear neutrophils taken from patients with aggressive periodontitis and chronic periodontitis. MATERIAL AND METHODS: Peripheral blood polymorphonuclear neutrophils obtained from 12 patients with chronic periodontitis, six patients with aggressive periodontitis and 12 healthy controls were exposed to P. gingivalis and A. actinomycetemcomitans following opsonization of the bacteria using the patient's own serum. Serum immunoglobulin G (IgG) levels against both periodontopathogens were measured. Phagocytosis and killing of the bacteria, as well as the extracellular human neutrophil elastase activity, were quantified. The total amount and the extracellular release of reactive oxygen species were measured using luminol-dependent and isoluminol-dependent chemiluminescence. RESULTS: Polymorphonuclear neutrophils from patients with chronic (62.16 +/- 19.39%) and aggressive (43.26 +/- 26.63%) periodontitis phagocytosed more P. gingivalis than the healthy controls (24.43 +/- 19.87%) at the 30-min time point after exposure to the bacteria (p < 0.05). High serum IgG levels against P. gingivalis and A. actinomycetemcomitans were detected in subjects with periodontitis. Polymorphonuclear neutrophils from subjects with chronic and aggressive periodontitis released significantly more reactive oxygen species and demonstrated greater human neutrophil elastase activity in the absence of any stimulus than polymorphonuclear neutrophils from healthy controls (p < 0.05). Polymorphonuclear neutrophils in chronic periodontitis released significantly more reactive oxygen species when exposed to P. gingivalis and A. actinomycetemcomitans than polymorphonuclear neutrophils in aggressive periodontitis. CONCLUSION: High serum IgG levels against P. gingivalis and A. actinomycetemcomitans promote phagocytosis in periodontitis. The extracellular release of reactive oxygen species and neutrophil elastase by polymorphonuclear neutrophils may also contribute to damage of the surrounding periodontal tissues.

Evidence for a specific crevicular lymphocyte profile in aggressive periodontitis.

BACKGROUND AND OBJECTIVE: It is undisputed that the periodontal pocket is a particular region of the host defense that is dominated by polymorphonuclear leukocytes. However, little is known about the lymphocytes in the crevice. It was the aim of this study to analyse the proportions of T cells (CD3+), T-helper cells (CD4+), T-suppressor cells (CD8+), and B cells (CD20+) in the crevice of patients with localized aggressive periodontitis (LAP), generalized aggressive periodontitis (GAP), and generalized chronic periodontitis (CP). The results were compared with those obtained from periodontally healthy controls. MATERIAL AND METHODS: Crevicular cells were collected according to a previously described method. The lymphocyte subpopulations were analysed by using an indirect immunofluorescence method. RESULTS: Significant differences were established between the test groups and the controls regarding the mean number of CD8+ lymphocytes (LAP > CP and controls; p < 0.05) and CD20+ lymphocytes (LAP/GAP > CP, p < 0.05 and LAP/GAP > controls; p < 0.001). Significant variations in the CD4+/CD8+ ratio were observed (LAP < controls and GAP < controls; p < 0.01), as well as a correlation between the number of T cells and the degree of inflammation. CONCLUSION: In the present study, patients with LAP and patients with GAP were found to have increased numbers of crevicular T-suppressor/cytotoxic and B cells. This supports the hypothesis of a changed immune pathology in patients with aggressive periodontitis.

Effect of smoking on crevicular polymorphonuclear neutrophil function in periodontally healthy subjects.

BACKGROUND: Polymorphonuclear neutrophils (PMNs) represent the first line of cellular defences in the gingival crevice. Smoking, as probably the most important environmental risk factor for periodontitis, has been shown to adversely affect many neutrophil functions. OBJECTIVE: The aim of this study was to investigate the influence of smoking on PMN numbers and function in periodontally healthy smokers and non-smokers. METHODS: Sixty subjects were recruited: 15 non-smokers, 15 light smokers (< 5 cigarettes/day), 15 moderate smokers (5-15 cigarettes/day) and 15 heavy smokers (> 15 cigarettes/day). Full mouth plaque index, sulcus bleeding index and probing depths were measured. Crevicular washings were obtained from all subjects to harvest PMNs. Numbers of PMNs, percentage viability, and percentage phagocytosis of opsonized Candida albicans were recorded. RESULTS: Mean plaque scores and probing depths were (non-significantly) increased in smokers compared to non-smokers. Mean sulcus bleeding index scores were significantly lower in moderate (0.10 +/- 0.10) and heavy (0.07 +/- 0.11) smokers compared to non-smokers (0.14 +/- 0.13) (p < 0.05). Compared to non-smokers (1.73 +/- 1.08 x 10(6)/ml), the numbers of PMNs were higher in light (1.98 +/- 0.96 x 10(6)/ml) and moderate (2.03 +/- 1.43 x 10(6)/ml) smokers and were lower in heavy smokers (1.68 +/- 1.18 x 10(6)/ml), though there were no significant differences in PMN counts between the groups (p > 0.05). Percentage viability of PMNs was significantly lower in light (77.6 +/- 7.8%), moderate (76.5 +/- 8.2%) and heavy (75.0 +/- 6.5%) smokers compared to non-smokers (85.5 +/- 6.0%) (p < 0.05). Furthermore, the ability of PMNs to phagocytose was significantly impaired in light (58.3 +/- 4.1%), moderate (51.9 +/- 2.33%) and heavy (40.9 +/- 3.5%) smokers compared to non-smokers (74.1 +/- 4.1%) (p < 0.05), with evidence of a dose-response effect. CONCLUSION: Cigarette smoking adversely affected PMN viability and function in this periodontally healthy population.

Grapefruit consumption improves vitamin C status in periodontitis patients.

OBJECTIVE: Previous studies demonstrate a relationship between a lack of vitamin C and increased risk of periodontal disease. In the present study we examine the vitamin C plasma levels and inflammatory measures in periodontitis patients before and after the consumption of grapefruit. SUBJECTS AND METHODS: Fifty-eight patients with chronic periodontitis were assigned to the test group (non-smokers n=21, smokers n=17) and a diseased control group (non-smokers n=11, smokers n=9). Furthermore, 22 healthy subjects were recruited to compare vitamin C plasma levels between periodontally diseased and healthy subjects. Clinical evaluations, including plaque index (PI), sulcus bleeding index (SBI), probing pocket depths (PPD) and plasma vitamin C levels, were performed at baseline, and after two weeks of grapefruit consumption. RESULTS: At baseline, we observed significantly reduced plasma vitamin C levels in the test group and diseased controls in comparison with the healthy controls. On principle, smokers showed lower levels of vitamin C (mean 0.39 +/- 0.17 mg dl(-1)) than non-smokers (mean 0.56+/-0.29 mg dl(-1)). After grapefruit consumption, the mean plasma vitamin C levels rose significantly in the test group compared to the diseased controls (non-smokers: 0.87+/-0.39 mg dl(-1), smokers: 0.74+/-0.30 mg dl(-1)). Furthermore the SBI was reduced in the test group (non-smokers: from 1.68+/-0.6 to 1.05+/-0.6, p<0.001), whereas PI and PPD were unaffected. CONCLUSION: The present results show that periodontitis patients are characterised by plasma vitamin C levels below the normal range, especially in smokers. The intake of grapefruit leads to an increase in plasma vitamin C levels and improves sulcus bleeding scores. Longer term studies are necessary to determine whether other periodontal outcomes improve with such supplementation especially in smokers.

Periodontal dressing (Vocopac) influences outcomes in a two-step treatment procedure.

OBJECTIVES: It is not clear if periodontal dressing influences the long-term results in a non-surgical treatment procedure. MATERIAL AND METHODS: The periodontal parameters (pre-baseline) of 36 patients with aggressive periodontitis were obtained before the patients were treated initially (1st step) by a dental hygienist, who completely removed the supra- and subgingival concrements. Baseline parameters were raised 3 weeks after the 1st step, before the 2nd therapy step was conducted. It consisted of a non-surgical procedure, which comprised a closed full-mouth manual root curettage (root planing), immediate systemic application of metronidazole, and the placement of a periodontal dressing (Vocopac, Voco). The patients were randomized to two test groups having their periodontal packs removed after 3-4 days (group 1, n=12) and 7-8 days (group 2, n=12), respectively and a control group (n=12) without periodontal dressing. Clinical parameters were raised again after 6 and 24 months. RESULTS: Six and 24 months later, changes in probing pocket depth (PPD) and probing attachment level (PAL) were observed in all three groups compared with baseline, but the difference was significant in group 2 only. In addition, group 2 showed a greater reduction in mean PPD and also a significantly greater gain of attachment in comparison with the controls. CONCLUSION: Wound dressing has a positive effect on clinical long-term results using a two-step non-surgical procedure. Moreover, removing the dressing after 7-8 days leads to clearly better results than removing it earlier.

Adherence of Streptococcus mutans to various restorative materials in a continuous flow system.

A continuous flow system was developed to evaluate the adhesion of Streptococcus mutans ATCC 25175 to filling materials (Ariston, Tetric, Dyract, Compoglass, Vitremer, Aqua Ionofil, Ketac Fil, amalgam, Galloy and ceramics as controls). Streptococcus mutans was added to saliva-coated test specimens, and a nutrient broth permanently supplied over a time period of 48 h and then the weight of plaque, the number and viability of the bacteria adhering to the materials were determined. The weights of artificial plaque on all filling materials tested were higher than those on ceramics, the highest values were measured on the glass-ionomers. The amount of plaque correlates with the surface roughness, whereas there was no correlation of the surface roughness with the number of colony-forming units (CFU) of S. mutans. The CFU of adhering S. mutans also depends on the viability of the bacteria. The plaque on Ketac Fil contained a high number of viable bacteria. The fluorides of glass-ionomers do not efficiently prevent the attachment and the viability of S. mutans.

A 2-step non-surgical procedure and systemic antibiotics in the treatment of rapidly progressive periodontitis.

BACKGROUND: In the last few years knowledge about periodontal infections has increased enormously, nevertheless practitioners are still seeking guidelines for suitable treatment concepts. METHODS: The aim of this study was to examine the effect of doxycycline, metronidazole, and clindamycin used adjunctively in a 2-step nonsurgical procedure in patients with rapidly progressive periodontitis (RPP). The first step included scaling, root planing, and polishing (SRP) in each quadrant using 4 to 5 visits. The second step included full-mouth enhanced root planing (RP) and wound dressing in 1 or 2 visits after SRP and the beginning of antibiotic therapy. Forty-eight patients (mean age 32.4 years) with generalized RPP, with an average of 16 sites with probing depths (PD) deeper than 8 mm, and high counts of Porphyromonas gingivalis were randomly assigned to 4 different groups: group 1 (doxycycline) n = 12, group 2 (metronidazole) n = 15, group 3 (clindamycin) n = 11, and group 4 (control group; no antibiotic treatment) n = 10. Clinical evaluations, including plaque index (PI), sulcus bleeding index (SBI), probing depth (PD), clinical attachment level (CAL), and bacteriological and crevicular cell sampling, were done at baseline (BL), 3 weeks after SRP, and 6 and 24 months after RP. RESULTS: After the first step (SRP), we observed an improvement of PI and SBI in all 4 groups, but did not see any statistically significant PD reduction 3 weeks after SRP compared to baseline. However, 6 and 24 months after the second step (RP) we observed a significantly greater reduction of PD in groups 2 and 3 and a significantly greater CAL gain in comparison to groups 1 and 4. After 24 months, the attachment level gain in group 1 and group 4 was less than 1.5 mm, and less than 1.0 mm in PD site categories 6 to 9 mm and >9 mm. PI showed no significant difference between the groups throughout the period after SRP until 24 months, compared to 3 weeks after SRP. SBI decreased most in the metronidazole and clindamycin groups. P. gingivalis and Actinobacillus actinomycetemcomitans were almost completely eradicated in these 2 groups 24 months after RP. In addition, the phagocytotic capacity of crevicular polymorphonuclear neutrophils was increased in groups 2 and 3 after the second step. CONCLUSIONS: The present results show that metronidazole and clindamycin are effective antibiotics when used adjunctively in a 2-step nonsurgical procedure of scaling and root planing in RPP patients.

Altered chemotactic behavior of crevicular PMNs in different forms of periodontitis.

BACKGROUND, AIMS: Polymorphonuclear neutrophils (PMN) are the predominant host defence cells in the gingival sulcus. Previous work demonstrates that the in vitro phagocytosis of crevicular cells in localised early onset periodontitis (LEOP) and generalised early onset periodontitis (GEOP) lesions is diminished. The present study extends this work by characterizing the chemotaxis function of crevicular fluid (CF) PMNs in various forms of periodontitis. METHODS: We investigated 7 patients with LEOP, 11 patients with GEOP, 12 patients with adult periodontitis (AP) and 2 age- and sex-matched healthy control groups. The two deepest sites of each quadrant in test and control subjects were selected for crevicular sampling. Chemotaxis was performed in a micro chamber (moist atmosphere, 5% CO2, 37 degrees C, 30 min) using N-formyl-methionyl-leucyl-phenylalanine (FMLP, 1 x 10(-7) mol FMLP/l) as a chemoattractant. The total chemotaxis was defined as the difference between the number of cells migrating towards FMLP minus the number of cells migrating towards PBS, counted in 20 randomly selected fields. Membranes were examined microscopically at 400 x magnification. RESULTS: The chemotactic activity in the adult periodontitis group was significantly higher compared to the age-related control group. However, we found a statistically significant reduction of chemotactic activity in LEOP and GEOP patients compared to the controls. CONCLUSIONS: These results indicate an increase of chemotactic activity from CF-PMN in patients with adult periodontitis, but on the other hand, a significant reduction of chemotactic responsiveness of these cells in LEOP and GEOP lesions.

[Adjuvant antibiotic therapy as a 2-step treatment concept in early-onset periodontitis: a strategy for eradication of bacteria that cause periodontitis]. / Adjuvante Antibiotikagabe in einem Zweischritt-Therapiekonzept für die früh beginnende Parodontitis. Eine Strategie zur Eradikation parodontopathogener Bakterien.

BACKGROUND AND OBJECTIVE: The periodontal region is a source of gram-negative bacterial infection. The pathogens involved have recently also been demonstrated in atheromatous plaques. They may increase the risk of myocardial infarction. In this study a strategy for eradicating periodontal bacteria and thus healing in patients with periodontal pockets and advanced destruction of alveolar bone was examined. PATIENTS AND METHODS: Initial periodontal status was documented in 36 patients with periodontitis (24 women, 12 men) who were then randomly assigned to one of three groups. Those in two of the groups were given either metronidazole or doxycycline orally as adjuvant treatment, while the third group received no antibiotics. Patients in all groups were treated according to a two-step procedure. In step 1, extensive supra- and subgingival plaques and concrements were removed. In step 2, root debridement and/or closed curettage of all pockets was undertaken in one visit, and the antibiotic given to the patients in groups 1 and 2. Results were assessed 3 weeks after the first step and 6 months, 2 and 4 years after step 2. RESULTS: The initial examination at 3 weeks revealed significant changes in all three groups with regard to the incidence of plaque and sulcus bleeding, but not regarding probing depth and attachment level. However, there were significant changes in probing depth and attachment level in all groups after step 2. Periodontal attachment was significantly improved at 2 and 4 years in the patients on metronidazole, but not those on doxycycline or no antibiotics. The greatest decrease in bleeding tendency was recorded in the metronidazole group. This group also had the greatest gain of new alveolar bone compared with the other two groups. Complete eradication of Porphyromonas gingivalis and Actinobacillus (Hemophilus) actinomycetemcomitans, important pathogenic bacteria that may have an atherogenic action, was obtained only in patients on metronidazole. CONCLUSION: The tissue-sparing two-step procedure brought about good clinical and radiological results, which can be significantly bettered by the addition of metronidazole, achieving eradication of pathogens involved in periodontal disease.

Red light kills bacteria via photodynamic action.

With the increase in the number of antibiotic resistant strains of microorganism, the search for alternative treatments of microbial infections becomes all the more important. We report a novel method for bacterial inactivation based on the optical excitation of the naturally occurring (endogenous) photosensitzing porphyrins by red light. In particular, the pathogenic Gram-positive porphyrin producing ATCC strains Propionibacterium acnes, Actinomyces odontolyticus and Porphyromonas gingivalis were investigated. Sensitive autofluorescence spectroscopy revealed that these bacteria naturally synthezise the fluorescent photosensitizer protoporphyrin IX. In addition, bacterial plaque samples of periodontitis patients were studied. Non-labeled fluorescent bacterial colonies were exposed to red light at 632.8 nm, 100 mW/cm2 light intensity and 360 J/cm2 energy density using a helium-neon laser. The survival rate after a single phototreatment with red light was found to be 0.58 +/- 0.09 in the case of Propionibacterium acnes, 0.30 +/- 0.04 in Actinomyces odontolyticus and 0.59 +/- 0.10 in Porphyrormonas gingivalis compared to non-exposed bacteria suspensions. No photoeffect was found for the bacterium Streptococcus mutans which exhibited no detectable porphyrin autofluorescence. Red-light exposed plaque samples of patients showed significant reduction of colony forming units by 50% as well as a pronounced photoeffect on the pigmented species Prevotella intermedia. Taken together, these results suggest the treatment with red light can be potentially employed as an therapeutic method to inactivate certain pathogenic strains of porphyrin producing bacteria without the use of external photosensitizers.

Early-onset and adult periodontitis associated with abnormal cytokine production by activated T lymphocytes.

There is growing evidence for an important role of the immune system in the pathogenesis of periodontitis. To further characterize the possible immunoregulatory dysfunction of peripheral blood mononuclear cells (PBMC) in periodontitis patients, we investigated functional aspects of PBMC from patients with early-onset periodontitis (EOP) and adult periodontitis (AP). Compared to controls, we observed decreased proliferative responses of PBMC from patients with EOP following stimulation with a mitogenic stimulus (phytohemagglutinin). To investigate whether this abnormality reflects a modulation in cytokine production, we measured the in vitro production of interleukin (IL)-3, IL-5, granulocyte macrophage colony-stimulating factor (GM-CSF), and interferon-gamma (IFN-gamma) by activated PBMC. PBMC in EOP patients expressed significantly decreased levels of IFN-gamma protein in response to mitogenic stimulation. Reduced IFN-gamma secretion was associated with decreased IFN-gamma and IL-2 mRNA expression in these cells, as well as decreased HLA-DR surface expression on monocytes. On the other hand, we observed significantly higher levels of IL-5 and GM-CSF in the same system using PBMC from AP patients. These were comparable to the levels observed for patients with allergic asthma. These data imply that EOP is associated with decreased Th1-like cytokine expression, and that the PBMC response from patients with AP is predominantly Th2/Th0 in nature.

[Inlays made from the glass ceramic Bioverit II. A clinical and scanning electron microscopic study]. / Inlays aus der Glaskeramik Bioverit II. Eine klinische und rasterelektronenmikroskopische Studie.

Inlays made of glass ceramics Bioverit II were proved just after inserting with a composite, and after 6 month's and one year. No marginal clefts were observed at clinical controls. The scanning study demonstrated excellent compound enamel-composite-glass ceramics just after inserting. A 15 to 20 microns deep furrow within the composite at most of the proved teeth was recognized one year later. The margins of enamel and inlays were yet covered with composite.

[The results of caries prevention in kindergartens]. / Ergebnisse der Kariesprävention in Kindergärten.

It was proved some years, if the program of the WHO for the oral healthy of preschool children could be realized through a complex caries prevention in 5 kindergartens of the town Jena. Since 1986 we found in 50 per cents of six years old children dentures free of caries after three years of caries prevention.

[Investigations of centrifugal penetration of the tooth root hard substances by hydrogen peroxide]. / Untersuchungen zur zentrifugalen Durchdringung der Zahnwurzelhartsubstanzen durch Wasserstoffperoxide.

After application of hydrogen peroxide into the root canal of extracted teeth of patients between the ages of 18 and 30 years the antiseptic could be detected on the root surface by means of iodometry and in tooth cross sections in dentine and cementum by means of staining reaction (benzidine-peroxidase solution). Out of the root canals which were apically not passable and coronally closed the first traces of the instilled 5% hydrogen peroxide appeared after 14 min 45 sec on the root surface. In the dentine a mean penetration depth of hydrogen peroxide up to 1.28 mm (apical root area), 1.45 mm (middle root third) and 1.95 mm (coronal root area) could be measured. Best results were obtained after rinsing/suction treatment of the root canal, especially in combination with low-frequency ultrasound.

[Antibacterial efficiency and toxicity of hydrogen peroxide and other antiseptics]. / Antimikrobielle Wirksamkeit und Toxizität von Wasserstoffperoxid und anderen Antiseptika.

The results suggest a relatively favourable relation between minimum bactericidal concentration (enterococci) and toxic agent concentration (chicken embryos) of hydrogen peroxide in comparison with chlorhexidin-digluconate, sodium-tosylchloramide and peracetic acid. For chlorhexidin-digluconate a somewhat more favourable relation between minimum bacteriostatic and toxic concentration of the agent was established.

[Efficiency of long-term dental treatment]. / Effektivität langjähriger stomatologischer Betreuung.

250 patients in long-term treatment (about 12, 4 years) were examined concerning their oral symptoms (DMF/T, CPITN, GPM, Plaque-index, Sulcus-bleeding-index of Mühlemann and Son), number of teeth, number of extracted teeth and teeth with endodontic treatment during the treatment period. These patients were compared with 250 patients, who were not under regularly treatment. There are clear differences between both groups. It can be concluded that long-term treatment with exact fillung therapy, endodontic and periodontic treatment is able to maintain teeth until a high age of life.

[Measurements of the mercury vapour concentration of the room air during dosing and trituration of amalgam by DMG 410]. / Messungen der Quecksilberdampfkonzentration der Raumluft während der Dosierung und Trituration von Amalgam mit dem DMG 410.

The testet mechanical amalgamators DMG 410 (VEB Dentalfabrik Treffurt) showed sufficient tightness. After dosing and trituration of 5 (during 30 min) and of 25 amalgam mixtures (during 2 hours) an increase of the mercury vapour concentration was measured from 0.003 mg Hg/m3 air of the room (blank value) to 0.008 mg Hg/m3 air of the room and to 0.018 mg Hg/m3 air of the room in an unventilated room. A significant decrease of the mercury vapour concentration could be established within 10 minutes by ventilation of the measurement room.

[Influence factors on progression and result of the treatment of periapical inflammation]. / Einflussfaktoren auf Verlauf und Ergebnis der Behandlung periapikaler Entzündungen.

The removal of the root canal infection was rendered more difficult with initially present communication of the root canal with the oral cavity, the involvement of relatively resistant microorganisms (enterococci, candida spec. and others) in the infection, and the extensive destructions of the periodontium. Some reduction of the chance of success could be established in case of extensive lesions of the marginal, lateral and apical periodontium after a control period of three years. This indicates the role of the relations between periodontium and endodontium for the result of the therapy of periapical inflammations.

[Sensitivity of microorganisms of infected root canals to disinfectants]. / Empfindlichkeit von Mikroorganismen aus infizierten Wurzelkanälen gegenüber Desinfektionsmitteln.

The bactericidal and growth inhibiting effects of disinfectants on Streptococcus and Candida strains were tested in vitro. Chlorhexidine, when used in tiny concentrations (0.0001-0.0039%) inhibits the growth of Streptococci. With Wofasteril 0.32%) all Streptococci and Candida were killed within a minute. Next to a good bacteria inhibitor (0.001-0.032%), a sufficient bactericidal effect on Streptococci is shown by hydrogen peroxide when used in a 4% concentration and warmed to 45 degrees Centigrade (degrees C).

[Escape of hydrogen peroxide and oxygen from the root canal]. / Untersuchungen zum Austritt von Wasserstoffperoxid und Sauerstoff aus dem Wurzelkanal.

In vitro investigations demonstrate that hydrogen peroxide, like the oxygen emitted from the root canal, escapes into the area around the root. The amount measurable was dependent on the concentration, the amount of time that the hydrogen peroxide was left in the root canal and the patency of the foramen apicale. The growth of the Staphylococcus aureus (SG 511) was restricted in the area around those roots whose root canal contained a 5% solution of hydrogen peroxide.