Gingival crevicular fluid and plasma oxidative stress markers and TGM-2 levels in chronic periodontitis.

OBJECTIVE: This study was aimed to evaluate the gingival crevicular fluid (GCF) and plasma transglutaminase-2 (TGM-2), total antioxidant capacity (TAC), total oxidant status (TOS), ferric reducing antioxidant power (FRAP) and thiobarbituric acid reactive substances (TBARS) in patients with chronic periodontal disease. MATERIALS AND METHODS: Twenty patients with chronic periodontitis (CP), 20 patients with gingivitis and 20 healthy subjects were enrolled in the study. Clinical periodontal parameters including probing depth, clinical attachment level, plaque index and papillary bleeding index were recorded. GCF and plasma levels of TGM-2, TAC, TOS, TBARS and FRAP were analyzed. RESULTS: GCF TGM-2 was significantly lower in CP group than in gingivitis patients (P=0.006). GCF FRAP in CP and gingivitis groups was significantly lower than in healthy subjects (P<0.001). Plasma FRAP level was lower in gingivitis group when compared to healthy subjects (P=0.003). There was no significant difference in GCF and plasma TAC, TOS, TBARS and plasma TGM-2 levels among the study groups (P>0.05). GCF TGM-2 level was positively correlated with GCF TAC and negatively correlated with CAL. CONCLUSIONS: Decreased FRAP in GCF and plasma indicating lower antioxidant status of CP patients might suggest the role of oxidative stress in periodontitis. GCF TGM-2 data might suggest that TGM2 is associated with stabilization of the extracellular matrix and wound healing in periodontium rather than gingival inflammation.

Gingival Crevicular Fluid and Plasma Levels of Transglutaminase-2 and Oxidative Stress Markers in Cyclosporin A-Induced Gingival Overgrowth.

BACKGROUND: Transglutaminase (TGM)-2 has been shown to contribute to fibrosis by extracellular matrix accumulation in some organs and is activated by intracellular reactive oxygen species. The aim of this study is to investigate levels of gingival crevicular fluid (GCF) and plasma TGM-2 and oxidative stress markers (OSMs) in cyclosporin A (CsA)-induced gingival overgrowth (GO). METHODS: The study enrolled 20 healthy (H) individuals; 20 patients with gingivitis (G); 20 CsA-medicated patients with GO (CsA GO+); and 20 CsA-medicated patients without GO (CsA GO-). GCF and plasma levels of TGM-2 were analyzed by enzyme-linked immunosorbent assay. Spectrofluorometry was used to analyze thiobarbituric acid reactive substance (TBARS); ferric-reducing antioxidant power (FRAP); total oxidant status (TOS); and total antioxidant capacity (TAC). RESULTS: GCF TGM-2 level was elevated in CsA GO+ compared with G (P = 0.048) and H (P = 0.001) groups. GCF TBARS level was elevated in CsA GO+ compared with other groups (CsA GO- group: P = 0.003; G group: P <0.001; and H group: P <0.001) and was higher in CsA GO- than in H (P = 0.048). GCF FRAP level was lower in CsA GO- than in H (P = 0.04). Both CsA GO+ and CsA GO- groups had lower GCF TOS levels than H (P <0.001 and P = 0.002) and G (P = 0.003 and P = 0.04). GCF TAC was higher in CsA GO+ than in H (P = 0.02). Plasma TGM-2 level was elevated in CsA GO+ compared with G (P = 0.048) and H (P = 0.002). Plasma FRAP level was higher in H and CsA GO- than in CsA GO+ (P = 0.008 and P = 0.02). CONCLUSIONS: CsA use significantly alters GCF and plasma levels of TGM-2 and OSMs. TGM-2 may contribute to CsA-induced GO in CsA-treated patients by changing GCF and plasma levels of OSMs. Further studies are needed to prove causality and its direction.

Interleukin-6 Family of Cytokines in Crevicular Fluid of Renal Transplant Recipients With and Without Cyclosporine A-Induced Gingival Overgrowth.

BACKGROUND: Interleukin (IL)-6 family of cytokines, including IL-6, oncostatin M (OSM), leukemia inhibitory factor (LIF), and IL-11, have fibrogenic features. The current study determines gingival crevicular fluid (GCF) levels of fibrosis-related IL-6-type cytokines in cyclosporine A (CsA)-induced gingival overgrowth (GO). METHODS: Eighty non-smokers were included (40 CsA-medicated renal transplant patients with GO [GO+; n = 20] or without GO [GO-; n = 20], 20 individuals with gingivitis, and 20 healthy participants). Probing depth and plaque, papilla bleeding, and hyperplastic index scores were recorded. GCF samples were obtained from the mesio-buccal aspects of two teeth. GCF IL-6, IL-1ß, OSM, LIF, and IL-11 levels were analyzed by enzyme-linked immunosorbent assay. RESULTS: The GO+ and GO- groups had higher IL-6 total amounts than the healthy group (P <0.008). IL-1ß total amounts in the GO+ group were significantly higher than in both the healthy and GO- groups (P <0.008). OSM total amount was elevated in the GO+ and GO- groups compared with both the gingivitis and healthy groups (P <0.008). All groups had similar LIF and IL-11 total amounts (P >0.008). Moderate positive correlations were detected among IL-6, IL-1ß, OSM, and IL-11 total amount in GCF and clinical parameters (P <0.05). CONCLUSIONS: IL-6 and OSM increases in GCF as a result of CsA usage or an immunosuppressed state irrespective of the severity of inflammation and the presence of GO. The IL-6 family of cytokines might not be directly involved in biologic mechanisms associated with CsA-induced GO. Lack of an association between assessed IL-6 cytokines and CsA-induced GO might indicate distinct effects of these cytokines on fibrotic changes of different tissues.

Periodontal disease and systemic diseases in an older population.

OBJECTIVE: To evaluate the relationship between older adults' medical and oral conditions and their self-reports of periodontal conditions with clinically obtained data. BACKGROUND: Concerns about oral health of elders and its association with systemic diseases have been gaining more attention. METHODS: A total of 201 older subjects were interviewed about their previous medical and dental histories and were asked to complete a health questionnaire. Each subject received full mouth exam, including counting number of natural teeth remaining, gingival (GI) and plaque index (PI), CPITN and denture status. RESULTS: Elders who completed health questionnaires had mean age of 62.5. Mean CPITN score was 1.62(± 1.12), PI was 1.57(± 1.48), and GI was 1.55(± 1.31). Women had higher prevalence of CVD and osteoporosis than men (p=0.008, p=0.0001, respectively). Subjects who reported bleeding upon brushing had higher PI and GI scores (p=0.03, p=0.05, respectively). Smokers were more likely to describe their periodontal tissues as unhealthy (72.3% vs. 27.7%, p=0.01), whereas self-reports of healthy vs. unhealthy gums did not differ between non-smokers. CONCLUSION: These findings suggest that a number of systemic conditions are associated with indicators of periodontal disease, and self-reports of oral conditions are independent of systemic diseases.

The effect of adjunctive chlorhexidine mouthrinse on GCF MMP-8 and TIMP-1 levels in gingivitis: a randomized placebo-controlled study.

BACKGROUND: The aim of the present study was to evaluate the effect of adjunctive chlorhexidine (CHX) mouthrinse on gingival crevicular fluid (GCF) MMP-8 and TIMP-1 levels in plaque-associated gingivitis. METHODS: A total of 50 gingivitis patients were included in the present study. In addition to daily plaque control, CHX group rinsed with CHX, while placebo group rinsed with placebo mouthrinse for 4 weeks. GCF samples were collected, and clinical parameters including plaque index, papillary bleeding index, calculus index and pocket depth were recorded at baseline and 4 weeks. GCF MMP-8 and TIMP-1 levels were determined by immunofluorometric assay (IFMA) and enzyme-linked immunosorbent assay (ELISA), respectively. RESULTS: In both groups, GCF MMP-8 levels of anterior and posterior sites at four weeks were not different from baseline (p > 0.05). There were no significant differences in GCF MMP-8 levels between the study groups at four weeks (p > 0.05). GCF TIMP-1 levels of anterior and posterior sites at four weeks were higher compared to baseline in both groups (p < 0.05). There was no significant difference in GCF TIMP level between the study groups at four weeks (p > 0.05). CONCLUSIONS: CHX usage had no significant effects on the GCF MMP-8 and TIMP-1 levels in plaque-associate gingivitis. However, daily plaque control resulted in the increase of GCF TIMP-1 levels regardless of CHX usage.

Gingival crevicular fluid and plasma acute-phase cytokine levels in different periodontal diseases.

BACKGROUND: The aim of the present study is to investigate gingival crevicular fluid (GCF) and plasma acute-phase cytokines, interleukin-1ß (IL-1ß), interleukin-6 (IL-6), interleukin-11 (IL-11), oncostatin M (OSM), and leukemia inhibitory factor (LIF) levels in patients with different periodontal diseases. METHODS: Eighty individuals were included in this study; 20 with chronic periodontitis (CP), 20 with generalized aggressive periodontitis (GAgP), 20 with gingivitis, and 20 classified as healthy (H). Probing depth, clinical attachment level, plaque index, and papilla bleeding index were recorded. Plasma and GCF IL-1ß, IL-6, IL-11, OSM, and LIF levels were analyzed by enzyme-linked immunosorbent assay. RESULTS: CP and GAgP groups had significantly higher GCF IL-1ß, IL-6, and IL-11 levels when compared with the H group (P <0.05). Conversely, GCF LIF levels of the CP and GAgP groups were lower than those of the H group (P <0.05). GCF OSM levels did not differ significantly among study groups. Plasma levels of all the cytokines studied were not significantly different among the study groups. CONCLUSIONS: Based on the present data, elevated IL-1ß, IL-6, and IL-11 GCF levels, but not plasma levels, are suggested as reliable inflammatory biomarkers in periodontal diseases. Decreased LIF levels in diseased groups might reflect the possible beneficial effects of LIF in the modulation of inflammatory response in gingiva.

Gingival crevicular fluid calprotectin, osteocalcin and cross-linked N-terminal telopeptid levels in health and different periodontal diseases.

AIM: The aim of the present study was to investigate gingival crevicular fluid (GCF) calprotectin, osteocalcin and cross-linked N-terminal telopeptide (NTx) levels in health along with different periodontal diseases. MATERIAL AND METHODS: Twenty chronic periodontitis (CP), 20 generalized aggressive periodontitis (G-AgP), 20 gingivitis and 20 healthy subjects were included. Probing depth, clinical attachment level, plaque index and papillary bleeding index was recorded. GCF calprotectin, osteocalcin and NTx levels were analyzed by enzyme-linked immunosorbent assay (ELISA). RESULTS: CP, G-AgP and gingivitis groups had higher GCF calprotectin total amount compared to healthy subjects (p< 0.008). CP and G-AgP groups had similar, but higher levels compared to gingivitis groups (p< 0.008). CP and G-AgP groups had lower GCF osteocalcin total amount compared to gingivitis and healthy groups (p< 0.008). CP group had higher GCF NTx but lower osteocalcin total amount and osteocalcin/NTx ratio than the G-AgP group (p< 0.008). CONCLUSIONS: Our results suggest that elevated GCF calprotectin levels play a role as a reliable inflammatory marker in the pathogenesis of periodontal disease. Fluctuating GCF levels of osteocalcin and NTx might point out to the abnormal bone turnover in periodontitis. Our data document for the first time the role of NTx in the pathogenesis of different periodontal diseases.

Antimicrobial effect of adjunctive use of chlorhexidine mouthrinse in untreated gingivitis: a randomized, placebo-controlled study.

The aim of this study was to examine the effectiveness of chlorhexidine mouthrinse (CHX) in addition to daily plaque control on subgingival microbiota in patients with untreated gingivitis. Fifty gingivitis patients were randomized to CHX or placebo groups. CHX group rinsed with 0.2% CHX, while placebo group rinsed with placebo mouthrinse for 4 weeks. Subgingival plaque samples were collected and plaque index (PI), papilla bleeding index (PBI), calculus index, and probing pocket depth (PPD) were recorded at baseline and at 4 weeks. The amounts of Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, Prevotella intermedia, Tannerella forsythia, Fusobacterium nucleatum, and total bacteria were detected by quantitative real-time PCR method. In the CHX group the total bacteria count was significantly reduced in posterior teeth at 4 weeks (p < 0.05), while no significant decrease was observed in the placebo group (p > 0.05). CHX mouthrinse as an adjunct to daily plaque control could be useful in the management of plaque-associated gingivitis and in reducing the subgingival total bacteria count especially in posterior teeth.

Gingival crevicular fluid osteocalcin, N-terminal telopeptides, and calprotectin levels in cyclosporin A-induced gingival overgrowth.

BACKGROUND: The aim of this cross-sectional study is to investigate gingival crevicular fluid (GCF) osteocalcin, cross-linked N-terminal telopeptide (NTx), and calprotectin levels in cyclosporin A (CsA)-induced gingival overgrowth (GO). METHODS: Forty medicated patients with CsA including 20 with GO (CsA GO+), 10 without GO (CsA GO-), 10 with GO and chronic periodontitis (CsA CP) and 60 patients with CP alone, 20 patients with gingivitis, and 20 healthy patients were enrolled. Probing depth, clinical attachment level, plaque index, and papillary bleeding index were recorded. GCF calprotectin, osteocalcin, and NTx levels were analyzed by enzyme-linked immunosorbent assay. Parametric tests were used for statistical analysis. RESULTS: The CsA GO+ and CP groups had significantly lower GCF osteocalcin levels and osteocalcin/NTx ratio than the healthy group, whereas GCF osteocalcin levels and osteocalcin/NTx ratio in the gingivitis group were higher than the CsA GO+, CsA GO-, CsA CP, and CP groups (P <0.05). The CP group had elevated GCF calprotectin levels compared to the other study groups (P <0.05). The CsA GO+ and CsA GO- groups also had higher GCF calprotectin levels compared to the CsA CP, gingivitis, and healthy groups (P <0.05). CONCLUSIONS: Increased GCF calprotectin and decreased GCF osteocalcin levels in the CsA GO+ and CsA GO- groups might suggest that CsA plays a role on the levels of these markers. The similarity of GCF osteocalcin, NTx, and calprotectin levels in the CsA GO+ and CsA GO- groups might suggest that these molecules are not involved in the pathogenesis of GO.

Toll like receptor 4 and membrane-bound CD14 expressions in gingivitis, periodontitis and CsA-induced gingival overgrowth.

OBJECTIVE: Immune cell recognition of lipopolysaccharides via CD14 and Toll-like receptor 4 (TLR4) complexes plays a crucial role in linking innate and adaptive immune responses. This study was aimed to investigate the expression of TLR4 and membrane-bound CD14 (mCD14) in the gingival tissues of patients with gingivitis, periodontitis and CsA-induced gingival overgrowth. DESIGN: Gingival tissues were obtained from 10 renal transplant patients receiving cyclosporine-A (CsA) and having gingival overgrowth (GO), 10 patients with chronic periodontitis, 10 generalized aggressive periodontitis, 10 gingivitis and 10 healthy subjects. Immunohistochemistry was performed in order to determine the localization of TLR4 and mCD14 in tissue specimens. RESULTS: TLR4 and mCD14 expressions were detected in all tissues including healthy gingival biopsies. TLR4 and mCD14 positive cells were predominantly confined to the epithelium-connective tissue interface area, and were highly expressed in the basal cell layer of patients with CsA GO and chronic periodontitis, compared to healthy group (P<0.05). CONCLUSION: The present study suggests that TLR4 and mCD14 protein expressions may be interrelated and appear to be associated with periodontal disease. CsA usage seemed not to affect TLR4 and mCD14 expressions in CsA induced GO tissues.

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.

Exclusion of candidate genes in seven Turkish families with autosomal recessive amelogenesis imperfecta.

Amelogenesis imperfectas (AI) are a group of inherited defects of dental enamel formation that show both clinical and genetic heterogeneity. Seven Turkish families segregating autosomal recessive AI (ARAI) were evaluated for evidence of a genetic etiology of AI for the seven major candidate gene loci (AMBN, AMELX, ENAM, FAM83H, KLK4, MMP20, and TUFT1). Dental and periodontal characteristics of the affected members of these families were also described. The mean scores of DMFS and dfs indices were 9.7 and 9.6, respectively. The mean PPD was 2.2 mm and the percentage of the sites with plaque and BOP were 87.8% and 72.4%, respectively. The exons and intron/exon junctions of the candidate genes were sequenced and no gene mutations were identified in any individuals. These findings support the existence of an additional gene(s) that are etiologic for ARAI in these families.

Effect of human platelet-derived growth factor-BB on attachment of periodontal ligament cells on root surfaces.

OBJECTIVE: To evaluate the effects of platelet-derived growth factor-BB (PDGF-BB) on the attachment of human periodontal ligament cells (HPLCs) on the root surfaces demineralized with different agents. METHODS: We performed this study at Ege University, Izmir, Turkey between 2005 and 2006. Eighty root slices were subjected to one of following treatments after root planing: 1) only root planing, 2) Platelet derived growth factor-BB (PDGF-BB), 3) citric acid demineralization, 4) citric acid demineralization + PDGF-BB, 5) tetracycline hydrochloric acid (T-HCl) demineralization, 6) T-HCl demineralization + PDGF-BB, 7) ethylenediamine tetra-acetic acid (EDTA) demineralization, and 8) EDTA demineralization + PDGF-BB. Human periodontal ligament cells were seeded on the root surfaces. Following the 2-hour incubation period, the number of cells was calculated by the colorimetric assay. Three slices from each group were processed for scanning electron microscopy. The number of attached cells was tested by analysis of variance (p=0.050). RESULTS: There were no significant differences among the groups with regard to the mean number of attached cells (p=0.843), which was highest in the fourth group, and lowest in the sixth group. CONCLUSION: Root planing is the most important treatment to make the diseased root surfaces biocompatible to HPLCs adherence. Application of PDGF-BB to root surfaces demineralized with citric acid may be advocated to enhance periodontal regeneration.

Oral rehabilitation of a patient with amelogenesis imperfecta.

Amelogenesis imperfecta is a hereditary disorder that causes defective enamel development in the primary and permanent teeth. Clinical treatment is important to address the esthetic appearance of affected teeth, reduce dentinal sensitivity, preserve tooth structure, and optimize masticatory function. The purpose of this case report was to describe the diagnosis, treatment planning, and dental rehabilitation of a patient with autosomal recessive amelogenesis imperfecta. The patient was followed for 5 years, and evaluation 3 years after restorations revealed no pathology associated with the rehabilitation. The patient's esthetic and functional expectations were satisfied.

Effect of platelet-rich plasma on gingival and periodontal ligament fibroblasts: new in-vitro growth assay

The use of platelet-rich plasma (PRP) as a source of growth factors is reported to be beneficial for periodontal regeneration. The aim of this study was to evaluate its effect on gingival and periodontal ligament fibroblast healing on a special growth assay designed by the working group. A wound with a 5 mm of diameter has been performed on periodontal ligament (PDL) and gingival fibroblast (GF) cell cultures. The cell wells were divided into five groups. The control group received only DulbeccoÆs modified EagleÆs medium/ HamÆs (DMEM) and the test groups received 0.5% PRP with 1/3 or 1/2 thrombin; 0.1% PRP with 1/3 or 1/2 thrombin. All of the groups were stained with haemotoxylene-eosine on days 2, 5, 7, 9 and 11. Digital screenings were performed on each time stop and the results were interpreted by means of % surface area covered by the cells. The results showed that 0.1% PRP with 1/3 thrombin group have closed up the wound circle in GF group at day 9 and in PDL group at days 9 and 11 with a significant difference when compared with other groups. GF response was significantly better than PDL cell response starting from day 5. Concluding, PRP favored wound closure in PDL and GF cell cultures and the developed growth assay may be utilized in future investigations of the biological basis of periodontal wound healing.