Impact of an Okinawa/Nordic based diet on endocrinological and periodontal conditions in individuals with type 2 diabetes. A randomized case-control study.

OBJECTIVES: To assess if the results following intake of a diet using an Okinawan-based Nordic diet (OBND) over one month differs in endocrinological, periodontal clinical outcome, and serum cytokine levels compared to a standard hospital care diet in individuals with diabetes type 2 (T2D) (control group). BACKGROUND: Scientific evidence suggests that the use of diet for individuals with T2D may be beneficial. METHODS: Participating individuals with T2D were randomly assigned to a test (OBND) (n = 14), or control group (n = 16). Anthropometric data, blood glucose levels, HbA1c levels, lipids, serum inflammation markers (CRP, and a routine panel of 24 cytokines), blood pressure, gingival bleeding on probing (BOP), probing pocket depths (PPD), and clinical attachment levels (CAL) were studied. RESULTS: Statistical analyses of baseline study data failed to demonstrate study group differences. The mean weight reduction was greater in the OBND group (4.1 kg) versus the control group (1.3 kg) (p < 0.01). The reduction in BMI was 1.4 kg/m2 in OBND (p < 0.001) and 0.5 kg/m2 in the control group, respectively (p < 0.01). Diastolic and systolic blood pressure reductions were greater in the OBND group than in the control group (p < 0.01). Periodontal study parameters (BOP % scores) and (PPD values) decreased (p < 0.001) overall with no between group differences. The OBND resulted in reduction of serum levels of IFNγ, Eotaxin IL-9, IP10,IL17a, MCP-1,m and PDFBB compared to the control diet. CONCLUSIONS: A strict T2D- diet provides an association between reduction in serum HbA1c and BOP scores. Serum levels decreases in IFNγ, Eotaxin IL-9, IP-10, IL17a. MCP-1, and PDFBB were only found in the test group.

Periodontal conditions, retinopathy, and serum markers in individuals with type 1 diabetes.

BACKGROUND: The prevalence of diabetes is high and increasing. Periodontitis has been identified as a risk factor in both type 1 and 2 diabetes. The study purpose was to assess periodontal conditions, retinopathy, and serum glutamic acid decarboxylase antibody (GADA) titers in relation to retinopathy in individuals with type 1 diabetes (T1D). METHODS: The study is a case series. Adult individuals with a diagnosis of T1D (n = 85) monitored ≥5 years were recruited from an endocrinology clinic. Peripheral venous blood samples were analyzed including assessments of serum HbA1c levels and GADA titers. Medical and periodontal conditions were examined, and the data assessed. Independent t tests, binary and multivariate analyses, χ2 and odds ratios were employed. RESULTS: Gingivitis was found in 68.2%, periodontitis in 21.2%, and retinopathy in 64.7%, GADA (≥35 U/mL) in 54.1%, and serum HbA1c > 48 mmol/mol in 94.3% of the individuals. The unadjusted odds ratio for periodontitis to differentiate a diagnosis of retinopathy was 7.3 (95%CI 1.6, 4.4, P <0.01). Multivariate analyses identified the following dependent factors to differentiate retinopathy; age, T1D duration, gingivitis, periodontitis at P < 0.001, sex, and serum GADA at P < 0.01, and by the number of remaining teeth at P < 0.05. CONCLUSION: Retinopathy as a complication to T1D is linked to the duration of diabetes, age of the individual and with increasing severity to periodontitis. Periodontal intervention studies are warranted.

Water and ethanol extracts of Plantago major leaves show anti-inflammatory activity on oral epithelial cells.

CONTEXT: The leaves of Plantago major have been used for the treatment of wounds and inflammation in folk medicine from prehistoric times. However there is no report on the use of P. major to treat inflammation in oral epithelial cell lines. OBJECTIVE: The present study was undertaken to reveal possible anti-inflammatory effects of Plantago major leaf extracts on oral epithelial cells in-vitro. MATERIALS AND METHODS: Water- and ethanol-based extracts of P. major leaves were prepared from freeze-dried plant material, and tested in-vitro using the oral epithelial cell line H400. The anti-inflammatory activity of P. major was tested against E. coli lipopolysaccharide (LPS) using the nuclear factor kappa beta (NF-kB) assay. RESULTS: Both the water- and the ethanol-based extracts, as well as a combination of the two extracts, showed anti-inflammatory activity. A concentration of 0.1 mg/mL (on dry weight basis) yielded the best results for all extracts. DISCUSSION AND CONCLUSION: The results show that synergistic effects of both polyphenols and water-soluble compounds (possibly polysaccharides) are responsible for anti-inflammatory activities of P. major.

Improved General and Oral Health in Diabetic Patients by an Okinawan-Based Nordic Diet: A Pilot Study.

Periodontal disease, periodontitis as well as the preceding gingivitis, has been associated with both obesity and diabetes. Studies have shown that diet changes can lead to a lower incidence of such inflammation. The aim of the present case series over four weeks was to study the effects on medical and dental conditions in patients with type 2 diabetes of the consumption of the Okinawan-based Nordic Diet (OBND®). Medical and dental examinations were performed to estimate the general health and gingivitis/periodontitis. Serum cytokine levels were assessed using Luminex technology. Eight of ten study participants completed the study. All participants lost weight (p = 0.012). Six out of seven that were treated with insulin could reduce their insulin intake after two weeks with OBND®. The reduction was about 16 units which corresponds to a 34% relative reduction compared to the starting point (range 15­63%). Fasting blood glucose values fell (p = 0.035). Hemoglobin A1c (HbA1c) (p = 0.01), triglycerides (p = 0.05), and low-density lipoprotein (LDL) (p = 0.05) were also reduced. Bleeding on probing changed from ~28% before any dietary changes to ~13% after two weeks with OBND® (p = 0.01). The reduction in gingival bleeding was as substantial as might be expected from one session of professional tooth cleaning. Markers of inflammation were also reduced. The OBND® thus showed significant promise in alleviating the impact of diabetes on dental as well as general health.

Cytokine and microbial profiles in relation to the clinical outcome following treatment of peri-implantitis.

AIM: To study whether cytokine levels and bacterial counts in p atients with peri-implantitis reflect clinical treatment outcome following non-surgical management. MATERIALS AND METHODS: Luminex magnet bead technology and checkerboard DNA-DNA hybridization were used to assess treatment outcome after treatment at the implant with the most severe peri-implantitis in 41 participants. RESULTS: Study group mean age was 40.3 years (SD ± 9.9). Stable treatment outcome after 6 months (no further bone loss, probing pocket depth decrease ≥0.5 mm, no bleeding/suppuration) was identified in 9 of 41 (22%) participants. Peri-implant crevicular fluid (PICF) levels were also lower for Il-1ß (P < 0.01), and with trends of lower cytokine levels in PICF for TNF-α (P = 0.071), PDGFBB (P = 0.071), as well as for VEGF (vascular endothelial growth factor) (P = 0.071), and bacterial counts for Actinomyces israelii, Aggregatibacter actonomycetemcomitans (Y4), Campylobacter gracilis, Echerichia coli, Fusobacterium periodonticum, Leptotrichia buccalis, Parvimonas micra, Staphylococcus haemolyticus, Streptococcus anginosus, and Tannerella forsythia. Increasing levels of IL-1 ß and S. aureus (r2  = 0.856) were found only at implants with non-stable outcome. A reduction of PICF levels for selected cytokines and bacteria studied had a sensitivity of 0.77, and a specificity of 0.80 against the clinical outcome as gold standard. Data analysis failed to differences in treatments (PerioFlow® versus YAG: ER laser) for changes in the expression of cytokines and bacteria studied. CONCLUSIONS: At 6 months, clinically stable treatment outcome of peri-implantitis is associated lower levels of putative pathogens total bacterial load with ≥30% reduction of IL1-ß, L-6, and VEGF levels in PICF.

Alteration of Neutrophil Reactive Oxygen Species Production by Extracts of Devil's Claw (Harpagophytum).

Harpagophytum, Devil's Claw, is a genus of tuberiferous xerophytic plants native to southern Africa. Some of the taxa are appreciated for their medicinal effects and have been traditionally used to relieve symptoms of inflammation. The objectives of this pilot study were to investigate the antioxidant capacity and the content of total phenols, verbascoside, isoverbascoside, and selected iridoids, as well as to investigate the capacity of various Harpagophytum taxa in suppressing respiratory burst in terms of reactive oxygen species produced by human neutrophils challenged with phorbol myristate acetate (PMA), opsonised Staphylococcus aureus, and Fusobacterium nucleatum. Harpagophytum plants were classified into different taxa according to morphology, and DNA analysis was used to confirm the classification. A putative new variety of H. procumbens showed the highest degree of antioxidative capacity. Using PMA, three Harpagophytum taxa showed anti-inflammatory effects with regard to the PBS control. A putative hybrid between H. procumbens and H. zeyheri in contrast showed proinflammatory effect on the response of neutrophils to F. nucleatum in comparison with treatment with vehicle control. Harpagophytum taxa were biochemically very variable and the response in suppressing respiratory burst differed. Further studies with larger number of subjects are needed to corroborate anti-inflammatory effects of different taxa of Harpagophytum.

Systemic inflammatory impact of periodontitis on acute coronary syndrome.

AIM: A causative relationship between acute coronary syndrome (ACS) and periodontitis has yet to be defined. The aim of this study was to assess differences in levels of serum cytokines between individuals with or without ACS or periodontal comorbidity. MATERIAL AND METHODS: In a case-control study, individuals with ACS (78 individuals, 10.3% females) and matching healthy controls (78 individuals, 28.2% females) were included. Medical and dental examinations were performed to diagnose ACS and periodontitis. Serum levels of cytokines were assessed, using Luminex technology. RESULTS: A diagnosis of periodontitis in the ACS and control group was diagnosed in 52.6% and 12.8% of the individuals, respectively. The unadjusted odds-ratio that individuals with ACS also had periodontitis was 7.5 (95% CI: 3.4, 16.8, p < 0.001). Independent of periodontal conditions, individuals with ACS had significantly higher serum levels of IL8 (mean: 44.3 and 40.0 pg/ml) and vascular endothelial growth factor (VEGF) (mean: 82.3 and 55.3 pg/ml) than control individuals. A diagnosis of periodontitis made no difference in serum cytokine expressions. CONCLUSION: Elevated serum levels of VEGF were associated with ACS. Serum cytokine expression in individuals with ACS is unrelated to periodontal conditions.

Probiotic supplements and debridement of peri-implant mucositis: a randomized controlled trial.

OBJECTIVE: The aim of this double-blind randomized placebo-controlled trial was to evaluate the effects of probiotic supplements in adjunct to conventional management of peri-implant mucositis. MATERIALS AND METHODS: Forty-nine adult patients with peri-implant mucositis were consecutively recruited after informed consent. After initial mechanical debridement and oral hygiene instructions, the patients received a topical oil application (active or placebo) followed by twice-daily intake of lozenges (active or placebo) for 3 months. The active products contained a mix of two strains of Lactobacillus reuteri. Patients were clinically monitored and sampled at baseline and after 1, 2, 4, 12 and 26 weeks. The clinical end-points were pocket-probing depth (PPD), plaque index (PI) and bleeding on probing (BOP). In addition, the subgingival microbiota was processed with checkerboard DNA-DNA hybridization and samples of gingival crevicular fluid (GCF) were analyzed for selected cytokines with the aid of multiplex immunoassays. RESULTS: After 4 and 12 weeks, all clinical parameters were improved in both the test and the placebo group. PPD and BOP were significantly reduced compared with baseline (p < 0.05), but no significant differences were displayed between the groups. The clinical improvements persisted 3 months after the intervention. No major alterations of the subgingival microflora were disclosed and the levels of inflammatory mediators in GCF did not differ between the groups. CONCLUSIONS: Mechanical debridement and oral hygiene reinforcement resulted in clinical improvement of peri-implant mucositis and a reduction in cytokine levels. Probiotic supplements did not provide added benefit to placebo.

Cytokine expression in peri-implant crevicular fluid in relation to bacterial presence.

AIM: The aim was to assess clinical inflammatory parameters, cytokine levels and bacterial counts in samples from implant crevicular fluid in cases with untreated peri-implantitis. MATERIAL AND METHODS: Several bacterial species known to up-regulate pro-inflammatory cytokines have been associated with peri-implantitis. The Luminex magnet bead technology was used to study cytokines in crevicular fluid. The checkerboard DNA-DNA hybridization method was used to study bacterial counts in samples from 41 implants (41 individuals). RESULTS: Profuse bleeding and suppuration was found in 25/41 (61.0%) of the implants. The reliability of duplicate cytokine processing was high. In the presence of profuse bleeding, higher pg/ml levels of IL-1ß (p = 0.02), IL-8 (p = 0.04), TNF-α (p = 0.03) and VEGF (p = 0.004) were found. Higher concentrations of IL-1ß were found in the presence of suppuration, and if Escherichia coli (p = 0.001) or Staphylococcus epidermidis (p = 0.05) could be detected. CONCLUSION: Profuse bleeding and/or suppuration in untreated peri-implantitis can be associated with higher concentrations of IL-1ß, IL-8, TNF-α and VEGF in peri-implant crevicular fluid. A higher concentration of IL-1ß in peri-implant crevicular fluid was found in samples that were positive for E. coli or S. epidermidis.

Consumption of bilberries controls gingival inflammation.

Bioactive molecules in berries may be helpful in reducing the risk of oral diseases. The aim of this study was to determine the effect of bilberry consumption on the outcome of a routine dental clinical parameter of inflammation, bleeding on probing (BOP), as well as the impact on selected biomarkers of inflammation, such as cytokines, in gingival crevicular fluid (GCF) in individuals with gingivitis. Study individuals who did not receive standard of care treatment were allocated to either a placebo group or to groups that consumed either 250 or 500 g bilberries daily over seven days. The placebo group consumed an inactive product (starch). A study group, receiving standard of care (debridement only) was also included to provide a reference to standard of care treatment outcome. Cytokine levels were assayed using the Luminex MagPix system. The mean reduction in BOP before and after consumption of test product over 1 week was 41% and 59% in the groups that consumed either 250 or 500 g of bilberries/day respectively, and was 31% in the placebo group, and 58% in the standard of care reference group. The analysis only showed a significant reduction in cytokine levels in the group that consumed 500 g of bilberries/day. A statistically significant reduction was observed for IL-1b (p=0.025), IL-6 (p=0.012) and VEGF (p=0.017) in GCF samples in the group that consumed 500 g of bilberries daily. It appears that berry intake has an ameliorating effect on some markers of gingival inflammation reducing gingivitis to a similar extent compared to standard of care.

Antibacterial activity of berry juices, an in vitro study.

OBJECTIVE: The aim of the present study was to evaluate in vitro antibacterial activities of blackcurrant and sea buckthorn juices on bacteria associated with gingival inflammation. MATERIALS AND METHODS: The growth of selected bacteria (Streptococcus mitis, Streptococcus mutans, Streptococcus sanguinis, Streptococcus gordonii, Staphylococcus aureus, Staphylococcus epidermidis and Pseudomonas aeruginosa) was studied in vitro on agar plates. The content of phenols in the different extracts was measured with HPLC-ESI-MS. RESULTS: The spectrometric analysis identified that the highest level of the single phenols studied was found for ferulic acid (113 µg/ml) in blackcurrant juice. Sea buckthorn contained low levels of selected phenols. Total bacterial inhibition for all bacterial species studied was found at 20% berry juice concentration with pH varying between 4.1-5.4. CONCLUSIONS: The present study identified that in vitro bacterial growth on agar plates was inhibited by blackcurrant and sea buckthorn juices and that low juice pH explains bacterial in vitro growth. This may have clinical implications in biofilm development, reducing the risks for both tooth decay and gingivitis.

Effects of Plantago major L. leaf extracts on oral epithelial cells in a scratch assay.

ETHNOPHARMACOLOGICAL RELEVANCE: The present study was undertaken to evaluate the effects from different leaf extracts of the traditional medicinal herb Plantago major L. (plantain) on cell proliferation and migration in vitro, as a test for potential wound healing properties. MATERIALS AND METHODS: Water and ethanol-based extracts were prepared from Plantago major fresh and dried leaves, and tested in vitro in a scratch assay with oral epithelial cells. RESULTS: The scratch assay produced reliable results after 18 h. Most of the tested extracts increased the proliferation/migration of the oral epithelial cells compared to the negative control. A concentration of 1.0 mg/mL (on dry weight basis) appears to be optimal regardless of type of extract, and among the alternatives, 0.1 mg/mL was always better than 10 mg/mL. Ethanol-based extracts with a concentration of 10 mg/mL had very detrimental effects on cell proliferation/migration. At the other two concentrations, ethanol-based extracts had the most beneficial effect, followed by water extracts of fresh leaves, ethanol plus water extracts of dried leaves and, finally, water extracts of dried leaves. CONCLUSIONS: This study suggests that both the water extracts and the more polyphenol-rich ethanol-based extracts of Plantago major leaves have medicinal properties. Further research is, however, needed to determine what compounds are responsible for the wound healing effects.

Mitochondria transmit apoptosis signalling in cardiomyocyte-like cells and isolated hearts exposed to experimental ischemia-reperfusion injury.

Ischemia-reperfusion (I/R) is a condition leading to serious complications due to death of cardiac myocytes. We used the cardiomyocyte-like cell line H9c2 to study the mechanism underlying cell damage. Exposure of the cells to simulated I/R lead to their apoptosis. Over-expression of Bcl-2 and Bcl-x(L) protected the cells from apoptosis while over-expression of Bax sensitized them to programmed cell death induction. Mitochondria-targeted coenzyme Q (mitoQ) and superoxide dismutase both inhibited accumulation of reactive oxygen species (ROS) and apoptosis induction. Notably, mtDNA-deficient cells responded to I/R by decreased ROS generation and apoptosis. Using both in situ and in vivo approaches, it was found that apoptosis occurred during reperfusion following ischemia, and recovery was enhanced when hearts from mice were supplemented with mitoQ. In conclusion, I/R results in apoptosis in cultured cardiac myocytes and heart tissue largely via generation of mitochondria-derived superoxide, with ensuing apoptosis during the reperfusion phase.