Probiotic strains of Lactobacillus brevis and Lactobacillus plantarum as adjunct to non-surgical periodontal therapy: 3-month results of a randomized controlled clinical trial.

OBJECTIVES: To determine if periodontitis patients benefit from treatment with Lactobacillus brevis and Lactobacillus plantarum strains, applied into periodontal pockets as gel and thereafter taken as lozenges, as an adjunct to scaling and root planing (SRP). MATERIALS AND METHODS: In a double-blind, randomized, placebo-controlled trial, 40 patients received scaling and root planing (SRP) in two sessions within 7 days. Patients then received either probiotic gel and lozenges (n = 20) or placebo (n = 20). The primary outcome variable was the number of diseased sites (DS: PD > 4 mm + BOP) at the 3-month re-evaluation. The effects of gender, age, probiotic therapy, presence of Porphyromonas gingivalis or Aggregatibacter actinomycetemcomitans, smoking, tooth being a molar and interdental location were evaluated using a multivariate multilevel logistic regression model. RESULTS: The number of DS after 3 months was similar in the test (Me = 8, IQR = 5-11) and control (Me = 5, IQR = 1-10) groups. Both groups showed substantial but equivalent improvements in periodontal parameters. The logistic regression showed higher odds for the healing of gingival bleeding (OR = 2.12, p = 0.048) and lower odds for the healing of DS (OR = 0.51; p < 0.001) in the probiotic group. CONCLUSIONS: Patients with periodontitis benefit from adjunctive use of probiotics containing L. brevis and L. plantarum in terms of reduction of gingival bleeding. However, adjunctive probiotics increase the number of persisting diseased sites with PD > 4 mm and BOP. CLINICAL RELEVANCE: The adjunctive use of probiotics containing L. brevis and L. plantarum strains in treating chronic periodontitis results in a higher number of residual diseased sites when compared with SRP + placebo; its use is therefore unfounded.

Oral health and systemic inflammatory, cardiac and nitroxid biomarkers in hemodialysis patients.

BACKGROUND: Periodontal diseases have systemic inflammatory effects and have been adversely associated with cardiovascular diseases, which are also the most frequent cause of death in the end-stage renal disease. The aim of this cross-sectional study was to investigate the oral health and serum biomarkers among the hemodialysis (HD) patients in Slovenia. MATERIAL AND METHODS: 111 HD patients were periodontally examined and their sera were assayed for C reactive protein (CRP), cardiac troponin T (TnT), nitrite/nitrate (NOx) and antibody levels to A. actinomycetemcomitans and P. gingivalis. The association of oral health with systemic response was analyzed with Kruskal-Wallis test, Fisher's exact test and multivariate linear regression. RESULTS: Bleeding on probing without periodontal pockets was present in 5.2%, calculus without periodontal pockets in 42.1%, shallow periodontal pockets in 39.5% and deep periodontal pockets in 13.2% of dentate patients. There were 28.8% edentulous participants. 63.1% of the patients had CRP levels higher than 3 mg/L and 34.2% higher than 10 mg/L. TnT was detectable in all participants, with 25.2% exhibiting levels higher than 100 ng/L. The median level of NOx was 43.1 µmol/L. Participants with higher CRP were more likely to be edentulous and have higher TnT levels. A direct association of oral health with TnT or NOx was not detected. CONCLUSIONS: HD patients in Slovenia have compromised oral health and increased serum inflammatory and cardiac biomarkers. Edentulousness was an independent predictor for the increased CRP, indicating a need for improved dental care to retain the teeth as long as possible.

Chronic ingestion of Porphyromonas gingivalis induces systemic nitric oxide response in mice.

INTRODUCTION: Porphyromonas gingivalis induces nitric oxide (NO) production in various cells, systemic NO elevation being expected in chronic oral challenge. METHODS: Groups of BALB/c mice were inoculated orally with either live P. gingivalis ATCC 33277 or sterile broth on days 0, 2 and 4, with or without later administration of the inducible nitric oxide synthase (iNOS) inhibitor 1400W. Plasma and tissues were harvested on day 42 for assays of tumor necrosis factor-alpha (TNF-alpha), nitrite and nitrate (NOx) and tissue NO, or histology and iNOS immunohistochemistry. RESULTS: No signs of gingivitis were observed, but plasma NOx was significantly elevated (P = 0.028) as was TNF-alpha (P = 0.079) in P. gingivalis-inoculated animals compared with controls, NOx being reduced when 1400W was used. NO production in organs showed a similar trend, with significant elevation in liver (P = 0.017) and kidneys (P = 0.027), whereas concomitant treatment of inoculated animals with 1400W caused significant reductions in NO in aorta (P = 0.008) and kidneys (P = 0.046). Sham-inoculated 1400W-treated animals had significantly increased plasma NOx (P = 0.004) and liver NO (P = 0.04). NOx in plasma correlated significantly with NO production in lungs (0.35, P = 0.032) and kidneys (0.47, P = 0.003). Immunohistochemistry demonstrated iNOS activity in many tissues in all groups. CONCLUSION: Repeated oral administration of P. gingivalis induced systemic NO and NOx production in mice, probably by activating iNOS as suggested by the response to 1400W.

Single gavage with Porphyromonas gingivalis reduces acute systemic nitric oxide response in mice.

INTRODUCTION: Porphyromonas gingivalis, an important periodontal pathogen, can also induce host responses in distant tissues. P. gingivalis induces nitric oxide (NO) production in immune system cells and non-immune system cells, therefore NO might be involved in an acute systemic host response. METHODS: Eighteen female BALB/c mice were perorally inoculated with 10(8) colony-forming units live P. gingivalis ATCC 33277. Plasma nitrite and nitrate (NOx) and NO production in lungs, aorta, heart, liver, spleen, kidneys, and brain were measured at intervals after inoculation and compared with levels in 11 control animals. RESULTS: NOx levels were significantly (P = 0.017) lower at 7, 13, and 25 h after P. gingivalis inoculation. A similar trend in NO production occurred in most tested organs, but never reached statistical significance. The correlation between NOx in plasma and NO in liver was positive (Spearman correlation coefficient = 0.81, P = 0.0025) and marginal for kidney (0.58, P = 0.059). CONCLUSION: Single peroral inoculation of mice with P. gingivalis reduces the acute systemic NO response. As NO is important for host defense, the reduction of NO levels after exposure is likely to delay the host response, increasing the chances that infection with P. gingivalis will become established.

In vivo study of liposomes as drug carriers to oral mucosa using EPR oximetry.

The purpose of this study was to select the best types of liposomes for use as drug carriers for topical treatment of oral mucosal lesions. Electron paramagnetic resonance (EPR) oximetry, using the paramagnetic probe lithium phthalocyanine, was used in vivo to measure the effects of a hyperemic drug, benzyl nicotinate (BN) which was incorporated into liposomes of varying size and composition. The liposomes were made from either hydrogenated or non-hydrogenated soy lecithin and mixed with polymethyl methacrylate ointment for application. EPR oximetry was used to measure the partial pressure of oxygen (pO2) in the oral mucosa before and after application of liposomes. It was found that the most pronounced changes of pO2 in oral mucosa and also the longest action of the drug occurred after the topical application of BN in multi-lamellar liposomes made from hydrogenated soy lecithin (p<0.0001). When these liposomes were applied to oral mucosa over 3 successive days it was found that pO2 increased the most on the first day, the effect gradually decreased following application on the second and third days. The duration of the resulting hyperemia was the longest on the second day (p<0.01). Among the examined carriers, multi-lamellar liposomes made from hydrogenated soy lecithin appear to be the most appropriate for local drug delivery to oral mucosa.

The permeability of human cementum in vitro measured by electron paramagnetic resonance.

The structure and permeability of cementum are changed during the course of periodontal disease. In this study, the transport of water-soluble, spin-labelled molecules through cementum was studied by electron paramagnetic resonance (EPR). Cementum samples cut from different parts of the root were classified into four different groups: (A) samples exposed to the oral environment, (B) samples exposed to the periodontal-pocket environment; (C) samples cut from periodontally involved teeth but not exposed to saliva or periodontal pocket and (D) samples from sound young teeth extracted for orthodontic reasons. In order to obtain undamaged cementum, a dentine layer was left on each sample. Two methods were used to measure the diffusion coefficients of spin-labelled molecules in cementum dentine samples. First, the method of one-dimensional EPR imaging (EPRI) was used to evaluate the penetration of spin-labelled molecules into the cementum/dentine structure. Second, the diaphragm-cell method was used to determine the diffusion coefficients of the labelled molecules through the cementum under steady-state conditions. The results indicate that the interface between cementum and dentine is a barrier to diffusion. A set of diffusion (D) and partition (K) coefficients to describe the molecular transport in cementum, barrier and dentine was generated from the experimental data of both methods. For cementum (c), the barrier (b) and dentine (d) these coefficients were: Dc= 10(-8)cm2/s, Db= 10(-10)cm2/s, Dd= 10(-6)cm2/s and K=0.1. For the particular periodontally involved and uninvolved teeth the value of the rate-limiting barrier was DbA= 0.3 +/- 0.03 x 10(-10)cm2/s, DbB= 1 +/-0.3 x 10(-10)cm2/s, DbC= 0.3 +/- 0.03 x 10(-10)cm2/s, DbD= 0.4 +/- 0.05 x 10(-10)cm2/s. The largest diffusion flux across the dental hard tissue was found in the samples that had been exposed to the pocket environment (3.1 +/- 0.2) x 10(-9)cm2/s (p < 0.01), which coincided with the permeability calculated from the data evaluated by EPRI. The transport of the labelled molecules into and through the cementum dentine samples depends on the structure of the dental hard tissues, which changes during the course of periodontal disease. Knowledge of molecular diffusion across the tooth cementum/dentine structure is likely to be important for planning new treatments for periodontal disease.

Effects of periodontal dressings on fibroblasts and gingival wound healing in dogs.

In the present study the effects of different commercially available periodontal dressings (Peripac, Barricaid, Fittydent, Reso-Pack and Myzotect-tincture) on fibroblast (V-79-379A) proliferation and survival were tested in vitro. Barricaid, Fittydent and Reso-Pack periodontal dressings have only small inhibitory effects on cell proliferation (83.3 +/- 9%, 71.6 +/- 8.7% and 87.3 +/- 4.5% of control after 48 h, respectively) in comparison with the great inhibitory effect of Myzotect-tincture (2.9 +/- 0.1%) and Peripac (33.7 +/- 11.4%) (p < 0.001). Barricaid was the only dressing where 41% of cells survived after exposure, while the other four dressings killed all the cells in 6 days. In addition, the healing of artificially created gingival wounds covered by Barricaid and Reso-Pack was followed for 7 days in 12 Beagle dogs. Histological evaluation of gingival tissue demonstrated that wounds covered by Reso-Pack showed the best epithelisation and vascularity and the least inflammatory reaction in first 4 days. Later the observed parameters were similar with those of wounds covered by Barricaid or without pack. The present results indicate that Peripac periodontal dressing and Myzotect-tincture showed the highest cytotoxicity to fibroblasts in vitro. From the histological observations in Beagle dogs Reso-Pack has been found to be the most suitable dressing, followed by Barricaid.

Diagnostic systems for assessing alveolar bone loss.

Radiological diagnostics serves as a basic monitoring technique for alveolar bone loss which is a severe consequence of periodontal disease. To evaluate efficacy of Conventional Visual Radiography (CVR), and to assess a complete clinical status, we had used two more diagnostic systems. These are Digital Subtraction Radiography (DSR) and Probing Pocket Depth (PPD). Experimental Periodontitis was studied in 20 beagle dogs based on the measurements taken in the beginning (baseline), and before (11th month) and after the medical treatment (12th month). Data analyses pointed out the same clinical trend, i.e. a significant bone loss prior to medical treatment and its recovery to the initial state. Differences in metrics and measurement errors could be identified as causes for discrepancies between the systems, but a relationship between the CVR and PPD is worth of further research, as these systems do not appear to be entirely compatible, but rather complementary to each other.

Postnucleation orbital implants.

A follow-up study of a group of 40 patients after the enucleation completed by the intra-orbital acrylic spherical magnetic implant (Roper-Hall style) is presented. The mean postoperative observation time is 3.3 years. Late complications were present in two patients: the extrusion of the implant in one of them was probably caused by his poor state due to chronic dialysis, while the traumatic luxation of the implant in the other was surgically repaired. Postoperative results in all other patients were satisfactory with the adequate orbital volume substitution and the ability of the prosthesis to twist and turn in any axis without fitting problems.

Periodontal disease, periodontal treatment and systemic nitric oxide in dogs.

Thirty-two client-owned dogs treated for periodontal disease were divided in group 1 if no periodontitis, group 2 if ≤25%, and group 3 if >25% of the teeth present were affected with periodontitis. Blood was tested before and 2 weeks after periodontal therapy for nitrosyl hemoglobin (HbNO), plasma nitrite/nitrate (NOx) and 3-nitrotyrosine (NT) levels. No HbNO was detected in any of the animals tested. There was no significant difference in the NOx plasma levels within each group or across the groups before and after the treatment, but a noticeable increase in NOx plasma levels was observed in group 3 after the treatment. Plasma NT was detected in only one third of the animals. NO levels varied greatly across individual dogs. The data are suggestive of an overall increase in systemic NO response 2 weeks after periodontal treatment in dogs with advanced periodontal disease, but the response is greatly individually-dependent.

[Causes and treatment of hypersensitivity in tooth root]. / Vzroki in zdravljenje preobcutljivosti zobne korenine.

The review article describes aetiology and treatment of dental root hypersensitivity. The three main theories of pain transduction from the exposed dentin surface to pain receptors in dental pulp are described. In addition, therapeutic compounds used to stimulate reparatory dentin formation and mineralization of peritubular dentin are described as well. The application of resins and adhesives on exposed dentin root surfaces is advocated in cases of chronic hypersensitivity. Finally, the authors emphasize the need for correct oral hygiene and early orthodontic treatment, if indicated, to prevent gingival recession and hypersensitivity of exposed root surfaces.

Local delivery of liposome-encapsulated superoxide dismutase and catalase suppress periodontal inflammation in beagles.

BACKGROUND, AIMS: The purpose of this study was to evaluate the influence of oxygen free radical scavengers on periodontal inflammation and healing process. METHOD: Experimental periodontitis was induced by elastic ligatures around premolars (P2, P3, P4) and 1st molars (M1) in the upper and lower jaws of 15 beagle dogs. 9 months after the beginning of the experiment, the ligatures were removed. After 3 weeks of stabilization period, all teeth were supragingivally scaled. The animals were divided into 3 groups of 5 dogs. The 1st group received a liposome-encapsulated superoxide dismutase (SOD), the 2nd group a liposome-encapsulated catalase (CAT) and the 3rd group received both enzymes encapsulated in liposomes. 4 treatment modalities were tested in each group; i.e., supragingival scaling only (1), supragingival scaling and enzymes (2), supra- and sub-gingival scaling and root planing (3) and supra- and sub-gingival scaling and root planing with subgingival application of enzymes (4). Enzymes were delivered subgingivally on a daily basis for a period of 6 weeks. Gingival index (GI), probing depth (PD), clinical attachment level (CAL), radiographic analysis and the histological evaluation were performed. RESULTS: Around the teeth with scaling and root planing followed by the application of SOD, the greatest suppression of gingival inflammation (GI = 1.8 +/- 0.1 before versus GI = 1.2 +/- 0.2 after treatment) (p<0.003), the smallest size of connective tissue infiltrate (5.5 +/- 4.3%) (p<0.01), the greatest reduction of PD (PD= 3.2 +/- 1.0 mm before versus 2.00.7 mm after treatment) (p<0.001), and gain of CAL (CAL=3.0 +/- 1.7 mm before versus CAL=2.4 +/- 1.1 mm after treatment) (p<0.001) were observed. In addition, radiographic analysis showed the greatest alveolar bone apposition in the group of teeth treated with scaling and root planing followed by subgingival application of SOD or both enzymes (p<0.001). CONCLUSION: In conclusion, we demonstrated that scaling and root planing with subgingival application of liposome-encapsulated SOD suppress peridontal inflammation on experimentally induced periodontitis in beagle dogs.

Changes in TGF-beta 1 levels in gingiva, crevicular fluid and serum associated with periodontal inflammation in humans and dogs.

Transforming growth factor-beta (TGF-beta) represents a family of polypeptide growth factors, involved in embryogenesis, inflammation, regulation of immune responses and wound healing. To determine whether TGF-beta contributes to the evolution of periodontal disease, we assayed TGF-beta levels in gingiva and crevicular fluid of patients with gingivitis and periodontitis. In parallel, TGF-beta was quantified in gingival fluid and serum of beagles with experimentally-induced periodontitis. Disease was monitored by several clinical parameters including Plaque Index, Gingival Index, probing depth, and epithelial attachment loss. Gingival tissues were obtained from 9 patients at the time of periodontal surgery, and gingival fluid samples were collected from an additional population of 10 periodontal patients. In 14 beagles, experimental periodontitis was induced and gingival fluids collected 6 months later. Fluid was collected by paper strips and volume measured by Periotron. Additionally, sera was collected before and 9 months after the ligature-induced periodontitis in 7 beagles. The levels of TGF-beta 1 were measured by ELISA. In the patients, a significantly higher concentration of TGF-beta 1 was observed both in the gingival tissues and fluid samples obtained from the sites with deeper periodontal pockets than in the less involved sites. In beagles, TGF-beta 1 levels measured in gingival fluid were elevated in moderate disease, declining in fluid samples obtained from the pockets during more advanced experimental periodontitis. Furthermore, with the progression of experimental periodontitis, a decrease in TGF-beta 1 occurred in the sera of the beagle dogs. These data suggest that TGF-beta 1 may play a rôle in the pathogenesis and diagnosis of periodontal disease, and that its actions can be further explored in an animal model.