Effect of gingivitis on azithromycin concentrations in gingival crevicular fluid

Background: Macrolide antibiotics yield high concentrations in inflamed tissue, suggesting that their levels in gingival crevicular fluid (GCF) could be increased at gingivitis sites. However, the increased volume of GCF associated with gingivitis could potentially dilute macrolides. To determine whether these assumptions are correct, the bioavailability of systemically administered azithromycin was compared in GCF from healthy and gingivitis sites. Methods: Experimental gingivitis was induced in one maxillary posterior sextant in nine healthy individuals. Contralateral healthy sextants served as controls. Participants ingested 500 mg azithromycin, followed by a 250‐mg dose 24 hours later. Four hours after the second dose, plaque was removed from experimental sites. GCF was collected from eight surfaces in both the experimental and control sextants and pooled separately. GCF samples were subsequently collected on days 2, 3, 8, and 15, and azithromycin content was determined by agar diffusion bioassay. Results: On days 2 and 3, the pooled GCF volume at experimental sites was significantly higher than at control sites (P <0.01), and the total azithromycin mass in 30‐second GCF samples pooled from experimental sites was significantly higher than at control sites (P <0.02). However, there were no significant differences in azithromycin concentration between the experimental and control pools at any point. Concentrations exceeded 7.3 μg/mL on day 2 and 2.5 μg/mL on day 15. Conclusion: Azithromycin concentrations are similar in GCF from gingivitis sites and healthy sites, suggesting that the processes that regulate GCF azithromycin concentration can compensate for local inflammatory changes.     

Comparison of Azithromycin and Amoxicillin Before Dental Implant Placement: An Exploratory Study of Bioavailability and Resolution of Postoperative Inflammation

Background: Studies suggest that a single prophylactic dose of amoxicillin reduces early implant complications, but it is unclear whether other antibiotics are also effective. This study compared the local antimicrobial and anti‐inflammatory effects resulting from a single dose of azithromycin or amoxicillin before surgical placement of one‐stage dental implants. Methods: Healthy adult patients requiring one‐stage dental implant placement were allocated randomly to receive either 2 g amoxicillin (n = 7) or 500 mg azithromycin (n = 6) before surgery. Peri‐implant crevicular fluid (PICF) samples from the new implant and gingival crevicular fluid (GCF) from adjacent teeth were sampled on postoperative days 6, 13, and 20. Inflammatory mediators in the samples were analyzed by immunoassay, and antibiotic levels were measured by bioassay. Results: On day 6, azithromycin concentrations in GCF and PICF were 3.39 ± 0.73 and 2.77 ± 0.90 μg/mL, respectively, whereas amoxicillin was below the limit of detection. During early healing, patents in the azithromycin group exhibited a significantly greater decrease in GCF volume (P = 0.03, analysis of variance). At specific times during healing, the azithromycin group exhibited significantly lower levels of interleukin (IL)‐6 and IL‐8 in GCF than the amoxicillin group and exhibited significantly lower levels of granulocyte colony stimulating factor, IL‐8, macrophage inflammatory protein‐1β, and interferon‐gamma‐inducible protein‐10 in PICF. Conclusions: Azithromycin was available at the surgical site for a longer period of time than amoxicillin, and patients taking azithromycin exhibited lower levels of specific proinflammatory cytokines and chemokines in GCF and PICF. Thus, preoperative azithromycin may enhance resolution of postoperative inflammation to a greater extent than amoxicillin.     

Azithromycin as an Adjunctive Treatment of Generalized Severe Chronic Periodontitis: Clinical,Microbiologic, and Biochemical Parameters

Background: This study examines the efficacy of azithromycin in combination with non‐surgical periodontal therapy on clinical and microbiologic parameters and gingival crevicular fluid (GCF) matrix metalloproteinases‐8 (MMP‐8) levels over 6 months in patients with severe generalized chronic periodontitis (CP). Methods: Twenty‐eight of 36 patients with severe generalized CP were included in this randomized, double‐masked, placebo‐controlled, parallel‐arm study. They were randomly assigned to azithromycin or placebo groups (500 mg, once daily for 3 days). Probing depth (PD), clinical attachment level, dichotomous presence or absence of supragingival plaque accumulation, and bleeding on probing were recorded. GCF samples were obtained from one single‐rooted tooth with PD ≥ 6 mm, whereas microbiologic samples were collected from two single‐rooted teeth with PD ≥ 6 mm. Microbiologic parameters were analyzed by quantitative real‐time polymerase chain reaction for Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, Tannerella forsythia, Fusobacterium nucleatum, Prevotella intermedia, and total bacteria. GCF MMP‐8 levels were determined by immunofluorescence assay. Results: Azithromycin and placebo groups demonstrated similar but significant improvements in all clinical parameters (P <0.05). A. actinomycetemcomitans, P. gingivalis, T. forsythia, P. intermedia, and total bacteria significantly decreased over the 6‐month period in both groups, whereas F. nucleatum was significantly reduced in all visits in the azithromycin group, with the levels also being lower compared with those of the placebo group (P <0.05). The azithromycin and placebo groups exhibited significant reduction in GCF MMP‐8 levels at the post‐treatment visit and at 2 weeks (P <0.05). Conclusion: On the basis of the present findings, it can be concluded that adjunctive azithromycin provides no additional benefit over non‐surgical periodontal treatment on parameters investigated in patients with severe generalized CP.     

Effects of treatment on soluble tumour necrosis factor receptor type 1 and 2 in chronic periodontitis

Aim: We reported that soluble tumour necrosis factor receptor type 2 (sTNFR2)/type 1 (sTNFR1) ratios in gingival crevicular fluid (GCF) decreased as the severity of chronic periodontitis (CP) increased. This study investigated the effects of the periodontal treatment on TNF‐α, sTNFR1 and R2 in GCF and serum of CP patients. Material and Methods: Thirty‐five serum and 90 GCF samples were obtained from 35 CP patients (23 non‐smokers and 12 smokers) at baseline and after treatment. The levels of TNF‐α, sTNFR1 and R2 in serum and GCF were quantified by enzyme‐linked immunosorbant assay. Results: No significant differences were found in the serum levels of TNF‐α, sTNFR1 and R2 and the ratio of sTNFR2/R1 between baseline and after treatment. After treatment, sTNFR1 and R2 levels in GCF of non‐smokers and smokers were significantly decreased compared with baseline. However, the sTNFR2/R1 ratio was significantly increased (non‐smoker: 0.56±0.03–0.84±0.03, p<0.0001; smoker: 0.59±0.06–0.85±0.04, p=0.0019). There were no significant differences between non‐smoking and smoking CP groups in serum and GCF. Conclusion: The ratio of sTNFR2/R1 in GCF significantly increased after treatment, and could be related to the clinical state of CP.     

Comparison of gingival crevicular fluid sampling methods in patients with severe chronic periodontitis

Background: The analysis of samplings from periodontal pockets is important in the diagnosis and therapy of periodontitis. In this study, three different sampling techniques were compared to determine whether one method yielded samples suitable for the reproducible and simultaneous determination of bacterial load, cytokines, neutrophil elastase, and arginine‐specific gingipains (Rgps). Rgps are an important virulence factor of Porphyromonas gingivalis, the exact concentration of which in gingival crevicular fluid (GCF) has not been quantified. Methods: GCF was sampled from four sites per patient (one sample per quadrant using two samples per method) in 36 patients with chronic periodontitis. One week later, the procedure was repeated with alternative methods. Variables determined were loads of Aggregatibacter actinomycetemcomitans (previously Actinobacillus actinomycetemcomitans) and P. gingivalis, levels of interleukin‐6 and ‐8, activity of neutrophil elastase, and level of Rgps. Results: The detected cytokine levels were higher using paper strips compared to paper points. Bacteria were found in similar loads from paper strips and paper points. Rgps were only detectable in high quantities by washing the periodontal pocket. The level of Rgps correlated with the load of P. gingivalis. Conclusions: The use of paper strips was suitable for the simultaneous determination of microbial and immunologic parameters. Obtaining GCF by washing can be useful for special purposes. The gingipain concentration in periodontal pockets was directly determined to be ≤1.5 μM. This value indicated that most of the substrates of these proteases by in vitro assays identified until now can be easily degraded in P. gingivalis–infected sites.     

Clinical,Microbial, and Immune Responses Observed in Patients With Diabetes After Treatment for Gingivitis: A Three‐Month Randomized Clinical Trial

Background: Although patients with diabetes are frequently affected by periodontitis, only a few investigations have focused on gingivitis in this at‐risk population. This randomized placebo‐controlled clinical trial compared the response to a gingivitis treatment protocol that combined mechanical procedures and daily use of an essential oil (EO) mouthrinse between patients with and without diabetes. Methods: The whole‐mouth periodontal probing depth (PD), gingival index (GI), and plaque index (PI) were monitored in gingivitis cases among systemically healthy patients (n = 60) or those with diabetes (n = 60) at baseline and 3 months after treatment. Levels of Porphyromonas gingivalis, Tannerella forsythia, Aggregatibacter actinomycetemcomitans, and total bacterial load were determined by a real‐time polymerase chain reaction in intrasulci plaque samples. The volume of gingival crevicular fluid (GCF) was quantified, and interleukin‐1β (IL‐1β) levels were determined in GCF samples. After a full‐mouth ultrasonic debridement, patients were randomly assigned to an EO or a placebo rinse for 90 days (40 mL/day). The data were analyzed through repeated‐measures analysis of variance and multiple comparisons Tukey tests (P <0.05). Results: GI was more severe in the diabetes group. Diabetes impaired GI and reduced GCF volume. PD, bacterial levels, and IL‐1β improved similarly in both systemic conditions. The adjunctive use of EO provided greater reductions of PI, GI, total bacterial load, T. forsythia, A. actinomycetemcomitans, and GCF volume. Conclusions: Response to gingivitis treatment in patients with diabetes can slightly differ from that in patients without diabetes. Daily use of an EO mouthrinse after ultrasonic debridement benefited patients with and without diabetes.     

Soluble Triggering Receptor Expressed on Myeloid Cells 1 (sTREM‐1) in Gingival Crevicular Fluid: Association With Clinical and Microbiologic Parameters

Background: Soluble triggering receptor expressed on myeloid cells 1 (sTREM‐1) belongs to the immunoglobulin superfamily and is involved in amplification of the inflammatory response to bacterial infection. This cross‐sectional study aims to investigate the levels of sTREM‐1 in gingival crevicular fluid (GCF) of individuals without periodontitis and with chronic periodontitis (CP) or generalized aggressive periodontitis (GAgP) and their association with the levels of key periodontal pathogens in subgingival plaque. Methods: GCF and subgingival plaque samples were obtained from healthy sites of participants without periodontitis (n = 20) and periodontitis sites of patients with CP (n = 22) and GAgP (n = 20). sTREM‐1 levels in GCF were measured by enzyme‐linked immunosorbent assay. Porphyromonas gingivalis, Treponema denticola, Tannerella forsythia, and Aggregatibacter actinomycetemcomitans levels in subgingival plaque were analyzed by quantitative real‐time polymerase chain reaction. Results: sTREM‐1 levels in GCF were higher in CP and GAgP than healthy sites by 3.6‐ and 4.4‐fold, respectively, with no significant differences between the two forms of periodontitis. Moreover, sTREM‐1 levels in GCF were positively correlated with site‐specific clinical periodontal parameters and levels of P. gingivalis, T. denticola, and T. forsythia, but not A. actinomycetemcomitans, in subgingival plaque. Conclusion: Increased GCF levels of sTREM‐1 at diseased sites and their positive correlation with clinical and microbiologic parameters strengthen the association of this inflammatory marker with periodontitis.     

Levels of interleukin-21 in patients with untreated chronic periodontitis

Background: A growing body of evidence suggested that interleukin (IL)‐21 enhances the effector phase during T‐cell responses. The aim of our study is to determine the levels of IL‐21 in periodontal sites from patients with chronic periodontitis and controls. Methods: The population studied consisted of 34 patients (15 with chronic periodontitis and 19 healthy patients). Twenty samples (10 gingival crevicular fluid [GCF] and 10 gingival biopsies) were collected from each group before the patients with periodontitis received periodontal treatment. Total protein concentrations were measured in all samples; the presence of IL‐21 was confirmed by immunohistochemistry and Western blot, and IL‐21 levels were quantified through an enzyme‐linked immunosorbent assay. Statistical analyses were performed using statistical software. Data were expressed as patient means ± SDs or medians (interquartile ranges) by using the χ², Student t, and Mann‐Whitney U tests. Results: GCF IL‐21 was mainly detected in patients with chronic periodontitis (P <0.05). Levels of IL‐21 in gingival tissues were significantly higher in patients with chronic periodontitis compared to healthy individuals (P <0.05). The Western blot and immunohistochemical staining confirmed the presence of IL‐21 in periodontal tissues and GCF. Conclusion: IL‐21 was highly expressed in patients with chronic periodontitis, especially in gingival biopsies; therefore, IL‐21 might play a role in the T‐cell response.     

Effect of azithromycin, as an adjunct to nonsurgical periodontal treatment, on microbiological parameters and gingival crevicular fluid biomarkers in generalized aggressive periodontitis

Emingil G, Han B, Özdemir G, Tervahartiala T, Vural C, Atilla G, Baylas H, Sorsa T. The effect of azithromycin, as an adjunct to nonsurgical periodontal treatment, on microbiological parameters and gingival crevicular fluid biomarkers in generalized aggressive periodontitis. J Periodont Res 2012; 47: 729–739. © 2012 John Wiley & Sons A/S Background and Objective: To study the effectiveness of azithromycin in combination with nonsurgical periodontal therapy on clinical and microbiological parameters, and on the MMP‐8 and TIMP‐1 levels in gingival crevicular fluid, over a 6‐mo time‐period in patients with generalized aggressive periodontitis. Material and Methods: Thirty‐two patients with generalized aggressive periodontitis were included in this randomized, double‐blind, placebo‐controlled, parallel‐arm study. They were randomly assigned to azithromycin or placebo groups (500 mg once daily for 3 d). Probing depth, clinical attachment levels, presence of bleeding on probing and plaque were recorded. Gingival crevicular fluid samples were obtained from one single‐rooted tooth, while microbiological samples were obtained from two single‐rooted teeth, all with a probing depth of ≥ 6 mm. Microbiological parameters were analyzed by quantitative real‐time PCR for Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, Tannerella forsythia, Fusobacterium nucleatum, Prevotella intermedia and total bacteria. Gingival crevicular fluid biomarkers were determined by immunofluorometric assay and ELISA. Results: All clinical parameters improved, and microbiological parameters and gingival crevicular fluid MMP‐8 levels significantly decreased, over the 6‐mo period (p < 0.05); both groups demonstrated similar improvements. The azithromycin group presented a higher percentage of deep pockets resolved (probing depth reduction of ≥ 3 mm from baseline) compared with the placebo group at 1 mo (p < 0.05). Conclusion: Adjunctive azithromycin therapy provides no additional benefit over nonsurgical periodontal treatment on clinical parameters, microbiological parameters and gingival crevicular fluid biochemical markers investigated in patients with generalized aggressive periodontitis.     

Gingival crevicular fluid and serum matrix metalloproteinase-8 and tissue inhibitor of matrix metalloproteinase-1 levels in renal transplant patients undergoing different immunosuppressive therapy

Aim: We investigated gingival crevicular fluid (GCF) and serum matrix metalloproteinase‐8 (MMP‐8) and tissue inhibitor of matrix metalloproteinase‐1 (TIMP‐1) levels from renal transplant patients receiving cyclosporine‐A (CsA) and having gingival overgrowth (GO), from patients receiving CsA therapy and having no GO and patients receiving tacrolimus therapy. Material and Methods: GCF samples were collected from sites with GO (GO+) and without GO (GO?) in CsA patients having GO; and GO? sites in CsA patients having no GO; sites from tacrolimus, gingivitis and healthy subjects. GCF and serum MMP‐8 and TIMP‐1 levels were determined by a time‐resolved immunofluorometric assay (IFMA) and enzyme‐linked immunosorbent assay. Results: GO+ sites in CsA patients having GO had elevated GCF MMP‐8 levels compared with those of CsA patients having no GO, tacrolimus and healthy subjects (p<0.005), but these levels were similar to those of gingivitis. The GCF MMP‐8 level was higher in GO+ compared with GO? sites in CsA patients having GO (p<0.05). GCF TIMP‐1 levels were similar between groups. Tacrolimus patients had lower GCF MMP‐8 levels than gingivitis (p<0.005), but levels similar to the healthy group. Conclusion: These results show that CsA and tacrolimus therapy has no significant effect on GCF MMP‐8 levels, and gingival inflammation seems to be the main reason for their elevations.     

Gingival crevicular fluid and serum visfatin concentration: their relationship in periodontal health and disease

Background: Visfatin is a pleiotropic mediator that acts as growth factor, cytokine, and enzyme involved in energy including nicotinamide adenine dinucleotide metabolism and was recently demonstrated to exert several proinflammatory functions. The purpose of this study is to determine the presence of visfatin in gingival crevicular fluid (GCF) and serum samples and to find out their association, if any. Methods: At the beginning of the study, 40 individuals (20 males and 20 females; age range: 23 to 53 years) were selected and divided into three groups based on the gingival index, probing depths, clinical attachment levels, and radiologic parameters (bone loss). Group 1 (10 patients with healthy periodontium; age range: 25‐36 years), group 2 (15 patients with gingivitis; age range: 25‐36 years), and group 3 (15 patients with chronic periodontitis; age range: 23‐53 years) GCF (by microcapillary pipettes) and serum (by venipuncture) samples were collected to estimate levels of visfatin using an enzyme‐linked immunosorbent assay kit. Results: Mean visfatin concentrations increased in GCF and serum with the severity of disease from healthy to gingivitis to periodontitis groups and differed significantly (P <0.05). However, it was found that GCF values were higher than serum values. The highest to lowest visfatin concentrations were found in groups 3 through 1, respectively. Visfatin in GCF and serum correlated positively with periodontal parameters in the chronic periodontitis group. Conclusions: The results suggest that GCF and serum visfatin concentrations increase with the severity of periodontal disease. Hence, visfatin values were considered an inflammatory marker in periodontal disease in GCF and serum. Visfatin also deserves further consideration as a therapeutic target.     

TNF-alpha TGF-beta2 and IL-1beta levels in gingival and peri-implant crevicular fluid before and after de novo plaque accumulation

Aims: The aim of this split‐mouth study was to investigate levels of tumour necrosis factor alpha (TNF‐α), transforming growth factor beta (TGF‐β2) and interleukin‐1 beta (IL‐1β) in gingival crevicular fluid (GCF) and peri‐implant crevicular fluid (PICF) after a 21‐day‐period of de novo plaque accumulation in the same patient. Material and Methods: In 25 patients, samples of GCF and PICF were collected in the sulcus of the tooth and of the implant after professional hygiene. After the no‐hygiene phase (21 days), second samples of GCF and PICF were taken. Third samples were collected after 69 days of re‐establishment oral hygiene techniques. The crevicular fluids were used to determine the volume and the levels of TNF‐α, TGF‐β2 and IL‐1β. Results: The volume of the crevicular fluids increased significantly after 21 days of plaque accumulation around teeth and implants and decreased significantly by 69 days. TNF‐α and TGF‐β2 did not change significantly among the three different samples. A significant increase of IL‐1β was observed after plaque accumulation around the teeth GCF, whereas in the PICF the increase was not statistically significant. Conclusions: These data suggest that increased volumes of GCF and PICF could be useful markers of early inflammation in gingival and peri‐implant tissues. In the presence of de novo plaque, implants showed lower, and nearly significant, levels of IL‐1β compared with teeth.     

Periodontal Condition of Patients With Autoimmune Diseases and the Effect of Anti‐Tumor Necrosis Factor‐α Therapy

Background: The aim of this study is to evaluate the effect of autoimmune diseases (AIs), as well as anti‐tumor necrosis factor‐α (TNF‐α) therapy on the clinical and immunologic parameters of the periodontium. Methods: Thirty‐six AI patients (12 rheumatoid arthritis [RA], 12 psoriatic arthritis, and 12 systemic sclerosis) were recruited together with 12 healthy (H) and 10 RA patients receiving anti‐TNF‐α therapy (RA+). Periodontal indices including plaque index, gingival index (GI), probing depth (PD), and bleeding on probing (BOP) were measured, and gingival crevicular fluid (GCF) was collected from five deepest pockets using papers strips. The TNF‐α level was analyzed using enzyme‐linked immunosorbent assay. Analysis of variance test was used for statistical comparison between groups, whereas Pearson linear correlation coefficient test was used to examine the association between TNF‐α and periodontal status indices. Results: The three AI subgroups were very similar in clinical and immunologic parameters. GI was greater in the AI patients compared to the H and RA+ groups (1.91 ± 0.54, 1.21 ± 0.67, and 1.45 ± 0.30, respectively, P = 0.0005). AI patients exhibited significantly more BOP than H and RA+ (46.45% ± 17.08%, 30.08% ± 16.86%, and 21.13% ± 9.51%, respectively, P = 0.0002). PD in H and RA+ groups were lower than in the AI (3.47 ± 0.33, 3.22 ± 0.41, and 3.91 ± 0.49 mm, P = 0.0001). Number of sites with PD >4 mm was higher in AI patients compared to H and RA+ (42.44 ± 17.5 versus 24.33 ± 15.62 versus 33.3 ± 6.6, P = 0.0002). GCF TNF‐α was higher among the AI patients (1.67 ± 0.58 ng/site) compared to 1.07 ± 0.33 ng/site for the H group and 0.97 ± 0.52 ng/site for the RA+ group (P = 0.0002). A significant positive correlation was found between PD and TNF‐α levels in the GCF (r = 0.4672, P = 0.0002), BOP (r = 0.7491, P = 0.0001), and GI (r = 0.5420, P = 0.0001). Conclusions: Patients with AI diseases have higher periodontal indices and higher TNF‐α levels in GCF than H controls. Anti‐TNF‐α treatment appears to reverse this phenomenon.     

Is There an Interaction Between Polycystic Ovary Syndrome and Gingival Inflammation?

Background: The aim of this study is to evaluate the gingival crevicular fluid (GCF), saliva, and serum concentrations of tumor necrosis factor‐α (TNF‐α), TNF‐α receptor‐1 (TNF‐αR1), TNF‐αR2, and interleukin‐6 (IL‐6) in non‐obese females with polycystic ovary syndrome (PCOS) and either clinically healthy periodontium or gingivitis. Methods: Thirty‐one females with PCOS and healthy periodontium, 30 females with PCOS and gingivitis, and 12 systemically and periodontally healthy females were included in the study. GCF, saliva, and serum samples were collected, and clinical periodontal measurements, body mass index, and Ferriman‐Gallwey score (FGS) were recorded. Sex hormones, cortisol, and insulin levels were measured. TNF‐α, TNF‐αR1, TNF‐αR2, and IL‐6 were determined by enzyme‐linked immunosorbent assay. Kruskal‐Wallis followed by Bonferroni‐corrected post hoc Mann‐Whitney U tests were used to analyze the data. Results: The PCOS + gingivitis group revealed significantly higher GCF, saliva, and serum IL‐6 concentrations than the PCOS + healthy group (P <0.0001). The two PCOS groups exhibited significantly higher saliva TNF‐α concentrations than the control group (P = 0.024 and P = 0.013, respectively). The FGS index was significantly higher in the PCOS + gingivitis group than the PCOS + healthy group (P = 0.030). The PCOS + gingivitis group revealed significantly higher insulin concentration than the PCOS + healthy and control groups (P = 0.014 and P <0.0001, respectively). Serum TNF‐α, TNF‐αRs, and serum, GCF, and salivary IL‐6 levels correlated with the clinical periodontal measurements. Conclusions: PCOS and gingival inflammation appear to act synergistically on the proinflammatory cytokines IL‐6 and TNF‐α. Thus, PCOS may have an impact on gingival inflammation or vice versa. Additional studies are warranted to clarify the possible relationship between PCOS and periodontal disease.     

The combination of amoxicillin and metronidazole improves clinical and microbiologic results of one-stage, full-mouth, ultrasonic debridement in aggressive periodontitis treatment

Background: The aim of the present study is to assess clinical, microbiologic, and immunologic benefits of amoxicillin/metronidazole (AM) when performing full‐mouth ultrasonic debridement (FMUD) in generalized aggressive periodontitis (GAgP) treatment. Methods: Twenty‐four GAgP patients were divided into two groups: the FMUD group (n = 12), which received FMUD plus placebo, and the FMUD+AM group (n = 12), which received FMUD and 375 mg amoxicillin plus 250 mg metronidazole for 7 days. The following clinical outcomes were tested: plaque and bleeding on probing indices, pocket probing depth (PD), relative gingival margin position (GMP), and relative clinical attachment level (CAL). Total amount of Porphyromonas gingivalis (Pg), Aggregatibacter actinomycetemcomitans (Aa), Tannerella forsythia (Tf), and gingival crevicular fluid (GCF) concentration of interleukin (IL)‐10 and IL‐1β were also determined. All clinical, microbiologic, and immunologic parameters were assessed at baseline and at 3 and 6 months post‐therapy. The ANOVA/Tukey test was used for statistical analysis (α = 5%). Results: Amoxicillin/metronidazole used as an adjunct to the FMUD protocol added clinical and microbiologic benefits to GAgP treatment (P <0.05). FMUD+AM groups presented an additional PD reduction in initially deep PDs at the 3‐month follow‐up (3.99 ± 1.16 mm and 3.09 ± 0.78 mm for FMUD+AM and FMUD, respectively; P <0.05), a lower number of residual pockets at the 3‐ and 6‐month follow‐ups, and a statistical reduction in amounts of Aa (P <0.05). Analysis of Tf and Pg amounts, as well as IL‐10 and IL‐1β GCF concentrations failed to demonstrate a difference between the groups (P >0.05). Conclusion: It may be concluded that amoxicillin/metronidazole improves clinical and microbiologic results of FMUD in GAgP treatment.     

A Cross‐Sectional Assessment of Biomarker Levels Around Implants Versus Natural Teeth in Periodontal Maintenance Patients

Background: Recent studies point to the clinical utility of using peri‐implant sulcular fluid (PISF) as a valuable diagnostic aid for monitoring peri‐implant tissue health. The objectives of this study are to determine the levels of key biomarkers in PISF in periodontal maintenance participants and compare them with their corresponding levels in gingival crevicular fluid (GCF) obtained from the same participants. Methods: PISF and GCF were collected from an implant and a contralateral natural tooth after the clinical examination of 73 participants. The levels of interleukin (IL)‐1α, IL‐1β, IL‐6, IL‐8, IL‐10, IL‐12, IL‐17A, tumor necrosis factor (TNF)‐α, C‐reactive protein, osteoprotegerin, leptin, and adiponectin were determined using multiplex proteomic immunoassays. The correlation of biomarker concentrations between GCF versus PISF, within GCF or PISF, and with several covariates (age, brushing frequency, days since professional cleaning, probing depth [PD], and plaque index) were also determined. Results: Significantly higher levels of IL‐17A (P = 0.02) and TNF‐α (P = 0.03) were noted in PISF when compared with their levels in GCF. Significant positive correlations were noted between the concentrations of cytokines in PISF versus their levels in GCF. Among the covariates, a significant positive correlation was noted between mean PDs around implants and levels of IL‐1β (P <0.05) and IL‐8 (P <0.05) in PISF. Conclusion: The results of this study point to the differential expression of specific biomarkers in GCF versus their levels in PISF in periodontal maintenance patients, which is critical information before establishing PISF as a diagnostic fluid to monitor peri‐implant health.     

Matrix metalloproteinase‐8 concentration in shallow crevices associated with the extent of periodontal disease

Objective: The aim of this study was to analyse the association between matrix metalloproteinase‐8 (MMP‐8) concentration in shallow, mostly non‐bleeding gingival crevices, and the extent of periodontal disease. Material and Methods: Plaque, bleeding on probing (BOP), probing pocket depth (PPD) and attachment level (AL) were assessed clinically in 48 patients with chronic periodontitis. MMP‐8 concentrations in gingival crevicular fluid (GCF) from four shallow (PPD3 mm), and four diseased sites and in serum, were measured by enzyme‐linked immunosorbent assay. Results: The mean concentration of MMP‐8 in GCF from shallow crevices was 11.8 ± 12.8 ng/ml and from diseased sites was 150.1 ± 91.8 ng/ml. In subjects with moderate to high plaque scores, a statistically significant association was found between MMP‐8 concentration from shallow crevices and the extent of AL4 mm (p=0.028) and AL6 mm (p<0.001). Conclusion: The above association between MMP‐8 concentration in shallow crevices and attachment loss provides a new aspect to future studies of MMP‐8 as a prognostic marker for periodontal disease.     

Proinflammatory and Anti‐Inflammatory Cytokines in Gingival Crevicular Fluid and Serum of Patients With Rheumatoid Arthritis and Patients With Chronic Periodontitis

Background: The aim of this study is to evaluate proinflammatory and anti‐inflammatory cytokine levels in gingival crevicular fluid (GCF) and serum of rheumatoid arthritis (RA) and chronic periodontitis (CP) patients to assess whether cytokine profiles distinguish patients with RA and patients with CP while using healthy patients as background controls. Methods: A total of 49 patients, 17 patients with RA (three males and 14 females; mean age: 47.82 ± 10.74 years), 16 patients with CP (10 males and six females; mean age: 44.00 ± 7.00 years), and 16 controls (eight males and eight females; mean age: 28.06 ± 6.18 years) were enrolled. Patients with RA were under the supervision of rheumatologists; 15 of the patients with RA were being treated with methotrexate–sulfasalazine combined therapy, and two of the patients were being treated with leflunomid therapy. Periodontal parameters (plaque index, gingival index, probing depth, and clinical attachment level) were recorded. Interleukin (IL)‐1β, IL‐4, IL‐10, and tumor necrosis factor‐α (TNF‐α) were determined in GCF and IL‐1β and IL‐10 in serum by enzyme‐linked immunosorbent assay. Results: There were significant differences found among RA, CP, and control groups for all periodontal parameters (P <0.05). The total amount and concentration of GCF IL‐1 β, IL‐4, IL‐10, and TNF‐α were similar in RA and CP patients (P >0.05). Although the total amount and concentration of serum IL‐10 was not significantly different among the groups (P >0.05), serum IL‐1β was significantly lower in the RA group compared to CP patients and controls and was higher in GCF of the RA group compared to the CP group. Conclusions: Although clinical periodontal disease parameters indicated more severe periodontal disease in CP compared to RA patients, immunologic evaluation did not reveal consistent results regarding proinflammatory and anti‐inflammatory cytokine levels. This might be a result of the use of non‐steroidal anti‐inflammatory drugs and rheumatoid agents by patients with RA.     

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.     

Analysis of matrix metalloproteinase (MMP‐8 and MMP‐2) activity in gingival crevicular fluid from children with Down’s syndrome

Yamazaki‐Kubota T, Miyamoto M, Sano Y, Kusumoto M, Yonezu T, Sugita K, Okuda K, Yakushiji M, Ishihara K. Analysis of matrix metalloproteinase (MMP‐8 and MMP‐2) activity in gingival crevicular fluid from children with Down’s syndrome. J Periodont Res 2010; doi: 10.1111/j.1600‐0765.2009.01214.x. © 2009 John Wiley & Sons A/S Background and Objective: High levels of colonization by periodontopathic bacteria and a high prevalence of chronic inflammatory periodontal disease have been reported in children with Down’s syndrome. Matrix metalloproteinases (MMPs) are mediators of extracellular matrix degradation and remodelling, and are deeply involved in the course of periodontal disease. To clarify the relationship between Down’s syndrome and periodontitis, we investigated levels of MMP‐2 and MMP‐8 in gingival crevicular fluid (GCF) and detection of periodontopathic bacteria from subgingival plaque. Material and Methods: Samples of GCF and plaque were isolated from central incisors. Levels of MMPs were evaluated by enzyme‐linked immunosorbent assay, and periodontopathic bacteria were detected by polymerase chain reaction. Results: Levels of MMP‐2 and MMP‐8 in Down’s syndrome patients were higher than those in healthy control subjects. In the Down’s syndrome group, increases in these MMPs were observed in GCF from patients with an oral hygiene index score of < 2 and in GCF from sites that were negative for bleeding on probing. The detection rate of periodontopathic bacteria in Down’s syndrome patients was higher than that in the control subjects. Matrix metalloproteinase‐2 levels in sites harbouring Porphyromonas gingivalis or Aggregatibacter (Actinobacillus) actinomycetemcomitans were lower than in those without these microorganisms. Conclusion: These results suggest an increase in MMP‐2 and MMP‐8 in Down’s syndrome patients, regardless of whether inflammation of periodontal tissue is present or not.