Ayse Nalbantsoy

Evaluation of the immunomodulatory properties in mice and in vitro anti-inflammatory activity of cycloartane type saponins from Astragalus species.

ETHNOPHARMACOLOGICAL RELEVANCE Astragalus roots are used to treat leukemia and for their wound healing properties in Southeast Anatolia-Turkey. MATERIALS AND METHODS In vivo studies to investigate the effects of two Astragalus saponins were carried out on the immune response cytokines by using six to eight weeks old male Swiss albino mice. The production of IL-1β, TGF-1β, TNF-α, IL-2, IL-4 and IFN-γ cytokines was determined by ELISA. The spleen and lymph nodes, isolated from the mice subjects, were analyzed to realize induction of the surface antigen productions for IL-2Rα (CD25) and CD69. In addition, their effects on the targets of inflammation such as NF κB, iNOS and NAG-1 were investigated in cell-based assays. RESULTS The results suggested that AST VII and Mac B had positive effect on Th1 cytokine release (IL-2 and IFN-γ), and suppression on Th2 cytokine production (IL-4). The immunohistochemical results exhibited induction of both IL-Rα (CD25) and CD69 surface receptors justifying the Th1 cytokine release. The compounds did not affect NF-κB or NAG-1 activity but iNOS activity was inhibited by Mac B with an IC(50) of 156 μg/ml. CONCLUSIONS The results show that Ast VII and Mac B create powerful immunoregulatory effects without the stimulation of inflammatory cytokines in mice, and have no significant effect on the inflammatory cellular targets in vitro.

Interleukin‐17 and interleukin‐18 levels in saliva and plasma of patients with chronic periodontitis

BACKGROUND AND OBJECTIVE This study was planned to investigate whether patients with chronic periodontitis exhibit different salivary and/or plasma concentrations of interleukin (IL)-17 and IL-18 compared with clinically healthy subjects. MATERIAL AND METHODS Whole saliva and blood samples, together with full-mouth clinical periodontal recordings, were obtained from 22 otherwise healthy untreated nonsmokers with chronic periodontitis and from 21 systemically and periodontally healthy control subjects. The concentrations of IL-17 and IL-18 in saliva and plasma were determined using ELISAs. RESULTS The healthy control group exhibited significantly lower values in all clinical periodontal measurements (p < 0.001). The salivary concentration of IL-17 was significantly lower, and that of IL-18 significantly higher, in patients from the chronic periodontitis group compared with healthy control subjects (p = 0.025 and p = 0.009, respectively). Plasma IL-17 and IL-18 concentrations were similar in the two study groups (p > 0.05). CONCLUSION Within the limits of the present study, it may be suggested that an elevated salivary IL-18 level in untreated nonsmoker chronic periodontitis patients has the potential to be a biomarker for periodontal tissue destruction.

Adjuvant effects of Astragalus saponins macrophyllosaponin B and astragaloside VII.

AIM OF THE STUDY The present study was undertaken to evaluate the hemolytic activities of two immunomodulator Astragalus saponins [Macrophyllosaponin B (MacB) from Astragalus oleifolius DC. and Astragaloside VII (Ast VII) from Astragalus trojanus Stev.], and their adjuvant potentials on the cellular and humoral immune responses of Swiss albino mice against BSA. MATERIALS AND METHODS The hemolytic activity of Mac B and Ast VII was determined using 0.5% rabbit red blood cell. For adjuvant activity, Swiss albino mice were immunized subcutaneously with BSA 100 μg alone or with BSA 100 μg dissolved in saline containing Ast VII (30, 60, 120 and 240 μg), Mac B (30, 60, 90 and 120 μg) or Freunds adjuvant on Days 1 and 15. Sera and splenocytes were collected 2 weeks after the last immunization for concanavalin A (Con A)-, lipopolysaccharide (LPS)- and BSA-stimulated splenocyte proliferation assay and measurement of BSA-specific antibodies in serum. RESULTS Mac B and Ast VII showed a slight hemolytic effect, with 0.42% and 0.54% values, respectively, at the highest concentration of 500 μg/ml. Mac B and Ast VII significantly enhanced the Con A-, LPS-, and BSA-induced splenocyte proliferation in the BSA-immunized mice especially at 120 and 240 μg (P<0.001), and 60, 90 and 120 μg (P<0.05, P<0.01 or P<0.001) doses, respectively. BSA-specific IgG, IgG1 and IgG2b antibody titers in serum were also significantly enhanced by Ast VII (120 μg), Mac B (90 μg) and Freunds as compared to the control group (P<0.01 or P<0.001). Moreover, the IFN-γ and IL-4 levels in the sera were detected using ELISA two weeks after the last immunization. Ast VII and Mac B were also found to stimulate IFN-γ production such as Freunds, two weeks after the last immunization at doses of 120 μg and 90 μg, respectively, as compared to the control. CONCLUSION Results show that Ast VII and Mac B generate important specific antibody and cellular response against BSA in mice, proving their potentials as a new class saponin adjuvant.

Effects of Menstrual Cycle on Periodontal Health and Gingival Crevicular Fluid Markers

BACKGROUND Fluctuations in sex steroid hormones, which are also noticeable through the menstrual cycle of women, may impact periodontal health. The aim of this study is to evaluate the effect of hormonal changes occurring in the menstrual cycle on gingival inflammation and the gingival crevicular fluid (GCF) levels of interleukin 6 (IL-6), prostaglandin E(2) (PGE(2)), tissue plasminogen activator (t-PA), and plasminogen activator inhibitor-2 (PAI-2). METHODS Twenty-five gingivitis patients and 25 periodontally healthy subjects having regular menstrual cycles were seen at menstruation (ME) (1 to 2 days of menstruation), ovulation (OV) (12 to 14 days), and premenstrual phases (PM) (22 to 24 days). GCF and saliva samples were collected and clinical parameters including plaque index and bleeding on probing were recorded at each menstrual phase. Salivary estrogen and progesterone levels were analyzed to determine exact menstrual cycle days. GCF levels of IL-6, PGE(2), t-PA, and PAI-2 were measured by enzyme-linked immunosorbent assay. RESULTS The percentages of sites with bleeding on probing were significantly higher in ME (60.85 +/- 18.36) and OV (58.92 +/- 25.04) than in the PM (40.12 +/- 20.10) phase in the gingivitis group (P <0.001; repeated measures analysis of variance), whereas it was similar for all phases in the healthy group (P >0.05; repeated measures analysis of variance). GCF levels of IL-6 were significantly elevated in gingivitis patients compared to healthy subjects in all phases (P = 0.004, P = 0.041, and P = 0.046 for ME, OV, and PM, respectively; Mann-Whitney U test). GCF levels of IL-6, PGE(2), t-PA, and PAI-2 were unchanged in different menstrual phases in both groups (P >0.05; Friedman test). CONCLUSION The present study suggests that changes in the sex steroid hormones during menstrual cycles might have a limited effect on the inflammatory status of gingiva, but GCF cytokine levels were not affected.

Saliva and Serum Levels of Pentraxin-3 and Interleukin-1β in Generalized Aggressive or Chronic Periodontitis

BACKGROUND Pentraxin-3 (PTX3) is a multifactorial protein involved in immunity and inflammation, which is rapidly produced and released by several cell types in response to inflammatory signals. The aim of the present study is to evaluate saliva, serum levels of PTX3, interleukin (IL)-1β in patients with generalized chronic periodontitis (CP) or aggressive periodontitis (AgP), and periodontally healthy individuals. METHODS A total of 94 participants (25 patients with AgP, 25 patients with CP, and 44 periodontally healthy individuals matched with AgP and CP groups) were recruited. Saliva and serum samples were collected. Clinical periodontal measurements were recorded. PTX3, IL-1β levels in serum, and saliva samples were determined by enzyme-linked immunosorbent assay. Data were tested statistically using Kruskal-Wallis, Mann-Whitney U, and Spearman ρ rank test. RESULTS Serum and saliva data were similar in CP and AgP groups. Saliva levels of IL-1β were significantly higher in the AgP and CP groups than controls (P <0.05). Salivary PTX3 levels were similar in the CP and control groups. Significantly higher salivary concentrations of PTX3 were detected in the AgP group than the control group (P <0.05). Saliva PTX3 levels correlated with plaque index and bleeding on probing in the CP group (P <0.05). Serum and saliva PTX3 levels correlated with those of IL-1β in the AgP group (P <0.05). CONCLUSIONS It may be suggested that PTX3 is related with periodontal tissue inflammation. Its salivary concentrations may have a diagnostic potential. Additional intervention and follow-up studies coupling PTX3 concentrations with microbiologic analysis would better clarify its role in periodontal diseases.

Interleukin-33 Levels in Gingival Crevicular Fluid, Saliva, or Plasma Do Not Differentiate Chronic Periodontitis

BACKGROUND This study investigates whether gingival crevicular fluid (GCF), saliva, and plasma levels of interleukin-33 (IL-33) can differentiate individuals with chronic periodontitis from individuals with healthy periodontium. METHODS GCF, whole saliva, and plasma samples together with full-mouth clinical periodontal recordings were obtained from 32 otherwise healthy, non-smoker chronic periodontitis individuals and 25 systemically and periodontally healthy, non-smoker individuals. IL-33 levels in the biofluid samples were determined by enzyme-linked immunosorbent assay. Data were tested statistically by Mann-Whitney U test. RESULTS The GCF concentrations of IL-33 were significantly lower in chronic periodontitis individuals than in healthy individuals (P <0.0001), whereas the total amounts in GCF samples were similar (P >0.05). The salivary and plasma contrations of IL-33 were indifferent in the two study groups (P >0.05). CONCLUSIONS According to the present findings, the GCF, saliva or plasma levels of IL-33 could not differentiate chronic periodontitis individuals and periodontally healthy individuals. Larger-scale intervention studies may better clarify this issue.

Salivary osteocalcin levels are decreased in smoker chronic periodontitis patients

OBJECTIVES  This study was planned to investigate whether smoker chronic periodontitis patients exhibit different salivary concentrations of C-telopeptide pyridinoline cross-links of type I collagen (ICTP) and osteocalcin (OC) compared to the non-smoker counterparts. METHODS Whole saliva samples, full-mouth clinical periodontal recordings were obtained from 33 otherwise healthy chronic periodontitis patients and 36 systemically, periodontally healthy control subjects. Chronic periodontitis patients and healthy control subjects were divided into smoker and non-smoker groups according to their self reports. Salivary ICTP, OC levels were determined by Enzyme-linked Immunoassays. RESULTS Healthy control groups exhibited significantly lower values in all clinical periodontal measurements (P < 0.001). Smoker periodontitis patients revealed similar clinical periodontal index values with non-smoker counterparts (P > 0.05). Chronic periodontitis patients exhibited significantly higher salivary OC levels than healthy controls (P < 0.05). Smoker periodontitis patients revealed lower salivary OC levels than non-smoker counterparts (P < 0.001). Log ICTP levels in non-smoker chronic periodontitis patients were higher than non-smoker controls (P < 0.05). Smoker healthy control group revealed higher log ICTP levels than non-smoker counterparts (P < 0.001). CONCLUSIONS Within the limits of this study, it may be suggested that suppression of salivary osteocalcin level by smoking may at least partly explain the deleterious effects of smoking on periodontal status.

Salivary and Plasma Levels of Toll-Like Receptor 2 and Toll-Like Receptor 4 in Chronic Periodontitis

BACKGROUND This cross-sectional study was planned to investigate whether patients with chronic periodontitis exhibit different salivary or plasma concentrations of Toll-like receptor (TLR) 2 and TLR4 compared to subjects who are clinically healthy. METHODS Whole saliva and plasma samples were obtained and full-mouth clinical periodontal measurements were recorded from 22 otherwise healthy patients with chronic periodontitis and 21 systemically and periodontally healthy control subjects. Salivary and plasma TLR2 and TLR4 levels were determined by enzyme-linked immunoassays. Data were tested statistically using Mann-Whitney U test. RESULTS The healthy group exhibited significantly lower values in all clinical measurements (P <0.001). The salivary TLR2 levels were similar in the two study groups (P >0.05). The patients with chronic periodontitis exhibited significantly higher salivary TLR4 (P <0.01) and plasma TLR2 and TLR4 levels (P <0.05). CONCLUSION The present findings support a hypothesis that inflammation increases expression of TLRs which leads to an increased detection of TLRs in saliva and plasma, which could be useful as a diagnostic test for periodontal diseases.

Mass spectrometry guided venom profiling and bioactivity screening of the Anatolian Meadow Viper, Vipera anatolica.

This contribution reports on the first characterization of the venom proteome and the bioactivity screening of Vipera anatolica, the Anatolian Meadow Viper. The crude venom as well as an isolated dimeric disintegrin showed remarkable cytotoxic activity against glioblastoma cells. Due to the rare occurrence and the small size of this species only little amount of venom was available, which was profiled by means of a combination of bottom-up and top-down mass spectrometry. From this analysis we identified snake venom metalloproteases, cysteine-rich secretory protein isoforms, a metalloprotease inhibitor, several type A2 phospholipases, disintegrins, a snake venom serine protease, a C-type lectin and a Kunitz-type protease inhibitor. Furthermore, we detected several isoforms of above mentioned proteins as well as previously unknown proteins, indicating an extensive complexity of the venom which would have remained undetected with conventional venomic approaches.

Is obesity a possible modifier of periodontal disease as a chronic inflammatory process? A case-control study.

BACKGROUND AND OBJECTIVE This cross-sectional case-control study was conducted to provide a comparative evaluation of clinical periodontal measurements, together with serum levels of certain bioactive peptides and inflammatory cytokines, in relation to obesity. For this purpose, clinical periodontal measurements and the levels of serum leptin, adiponectin, interleukin-6 (IL-6), C-reactive protein and soluble intercellular adhesion molecule-1 of obese female individuals and their nonobese counterparts were compared. MATERIAL AND METHODS Sixty obese (body mass index (BMI) > 30) and 31 nonobese (BMI < 30) female subjects were recruited for the present study. Before any periodontal intervention, serum samples were obtained and full-mouth clinical periodontal measurements were recorded at six sites per tooth. ELISA was used for the biochemical analysis. Data were tested statistically. RESULTS Clinical attachment level was significantly higher in the obese group compared with the nonobese control group (p < 0.05). Serum levels of leptin and IL-6 were significantly higher in the obese group (p < 0.05). BMI correlated with the serum levels of inflammatory molecules (p < 0.05), but not with clinical periodontal parameters, in the obese group. CONCLUSION In conclusion, obesity does not seem to have a prominent effect on clinical periodontal parameters but it does have many correlations with circulating inflammatory molecules. As suggested in the literature, increased levels of leptin and IL-6 in the obese group might be one explanation for a possible relationship between obesity and periodontal disease. A prospective study is warranted to clarify, in greater detail, the effects of obesity on periodontal health.