Eduardo Jorge Feres-Filho

Systemic Antimicrobials Adjunctive to a Repeated Mechanical and Antiseptic Therapy for Aggressive Periodontitis: A 6-Month Randomized Controlled Trial

BACKGROUND The purpose of this study is to compare the additional benefit of systemic antimicrobials versus placebos to a repeated mechanical instrumentation combined with comprehensive local chemical plaque control for the periodontal treatment of generalized aggressive periodontitis (GAgP). METHODS This was a 6-month randomized, double-masked, placebo-controlled clinical trial. All GAgP patients received full-mouth disinfection followed by staged scaling and root planing without (placebo group; n = 17) or with (test group; n = 18) systemic antimicrobials (500 mg amoxicillin [AMX] + 250 mg metronidazole [MET]; three times a day for 10 days). Clinical parameters were measured at baseline and 3 and 6 months post-therapy. Significant differences between groups at baseline were sought by using the Mann-Whitney U test, whereas comparisons over time were examined by using a general linear model repeated measures procedure. RESULTS Both groups demonstrated similar improvements in most parameters over time. The test group presented a greater mean probing depth (PD) reduction and clinical attachment level (CAL) gain at sites with initially moderate PD at 6 months (P <0.03). No differences were seen between groups regarding mean reductions and mean gains, respectively, for PD and CAL initially ≥7 mm. The test group presented a higher percentage of sites that improved ≥2 mm and ended up with PD ≤4 mm or a lower percentage of sites that worsened ≥2 mm and remained with PD >4 mm at 3 months (P <0.01). No differences were noticed between groups for these parameters at 6 months. CONCLUSION AMX + MET brought additional clinical effects to the repeated mechanical and antiseptic treatment of GAgP in a very short time (3 months), which tended to fade away over time (6 months).

Kinin Danger Signals Proteolytically Released by Gingipain Induce Fimbriae-Specific IFN-γ- and IL-17-Producing T Cells in Mice Infected Intramucosally with Porphyromonas gingivalis

Porphyromonas gingivalis, a Gram-negative bacterium that causes periodontitis, activates the kinin system via the cysteine protease R-gingipain. Using a model of buccal infection based on P. gingivalis inoculation in the anterior mandibular vestibule, we studied whether kinins released by gingipain may link mucosal inflammation to T cell-dependent immunity through the activation of bradykinin B2 receptors (B2R). Our data show that P. gingivalis W83 (wild type), but not gingipain-deficient mutant or wild-type bacteria pretreated with gingipain inhibitors, elicited buccal edema and gingivitis in BALB/c or C57BL/6 mice. Studies in TLR2−/−, B2R−/−, and neutrophil-depleted C57BL/6 mice revealed that P. gingivalis induced edema through the sequential activation of TLR2/neutrophils, with the initial plasma leakage being amplified by gingipain-dependent release of vasoactive kinins from plasma-borne kininogens. We then used fimbriae (Fim) Ag as a readout to verify whether activation of the TLR2→PMN→B2R axis (where PMN is polymorphonuclear neutrophil) at early stages of mucosal infection had impact on adaptive immunity. Analyzes of T cell recall responses indicated that gingipain drives B2R-dependent generation of IFN-γ-producing Fim T cells in submandibular draining lymph nodes of BALB/c and C57BL/6 mice, whereas IL-17-producing Fim T cells were generated only in BALB/c mice. In summary, our studies suggest that two virulence factors, LPS (an atypical TLR2 ligand) and gingipain, forge a trans-cellular cross-talk between TLR2 and B2R, thus forming an innate axis that guides the development of Fim-specific T cells in mice challenged intrabuccally by P. gingivalis. Ongoing research may clarify whether kinin-driven modulation of T cell responses may also influence the severity of chronic periodontitis.

Long-term evaluation of the antimicrobial susceptibility and microbial profile of subgingival biofilms in individuals with aggressive periodontitis

This study evaluates the antimicrobial susceptibility and composition of subgingival biofilms in generalized aggressive periodontitis (GAP) patients treated using mechanical/antimicrobial therapies, including chlorhexidine (CHX), amoxicillin (AMX) and metronidazole (MET). GAP patients allocated to the placebo (C, n = 15) or test group (T, n = 16) received full-mouth disinfection with CHX, scaling and root planning, and systemic AMX (500 mg)/MET (250 mg) or placebos. Subgingival plaque samples were obtained at baseline, 3, 6, 9 and 12 months post-therapy from 3–4 periodontal pockets, and the samples were pooled and cultivated under anaerobic conditions. The minimum inhibitory concentrations (MICs) of AMX, MET and CHX were assessed using the microdilution method. Bacterial species present in the cultivated biofilm were identified by checkerboard DNA-DNA hybridization. At baseline, no differences in the MICs between groups were observed for the 3 antimicrobials. In the T group, significant increases in the MICs of CHX (p < 0.05) and AMX (p < 0.01) were detected during the first 3 months; however, the MIC of MET decreased at 12 months (p < 0.05). For several species, the MICs significantly changed over time in both groups, i.e., Streptococci MICs tended to increase, while for several periodontal pathogens, the MICs diminished. A transitory increase in the MIC of the subgingival biofilm to AMX and CHX was observed in GAP patients treated using enhanced mechanical therapy with topical CHX and systemic AMX/MET. Both protocols presented limited effects on the cultivable subgingival microbiota.

Expression of metalloproteinases and their tissue inhibitors in gingiva affected by hereditary gingival fibromatosis: analysis of three cases within a family

BACKGROUND AND OBJECTIVE Hereditary gingival fibromatosis (HGF) is a benign disorder manifested by fibrous enlargement of keratinized gingiva. Evidence exists concerning the role of matrix metalloproteinases (MMPs) and their tissue inhibitors (TIMPs) in mediating normal and pathological processes, including HGF. Nevertheless, there are few and contradictory results on the analysis of MMPs and TIMPs transcripts in this pathology. MATERIAL AND METHODS We studied the expression of the transcripts encoding MMP-1, -2 and -9 and TIMP-1 and -2 in gingival biopsies, obtained from three cases of HGF within a family, by semi-quantitative reverse transcriptase-polymerase chain reaction analysis. Samples were also processed for gelatin zymography. RESULTS Except for MMP-9, most transcripts presented a higher level of expression in biopsies from HGF patients compared with control subjects. Accordingly, MMP-9 gelatinase activity was detected at low and similar levels among samples. Moreover, MMP-2 enzymatic activity was not detected at all. CONCLUSION The mRNA expression of MMP-1 and -2 and TIMP-1 and -2 does not explain the gingival overgrowth presented in these cases. In addition, it is suggested that the gene expression of those molecules in the course of HGF is regulated at the translational or post-translational level.