Smoking-induced subgingival dysbiosis precedes clinical signs of periodontal disease.

Smoking accelerates periodontal disease and alters the subgingival microbiome. However, the relationship between smoking-associated subgingival dysbiosis and progression of periodontal disease is not well understood. Here, we sampled 233 subgingival sites longitudinally from 8 smokers and 9 non-smokers over 6-12 months, analyzing 804 subgingival plaque samples using 16 rRNA sequencing. At equal probing depths, the microbial richness and diversity of the subgingival microbiome was higher in smokers compared to non-smokers, but these differences decreased as probing depths increased. The overall subgingival microbiome of smokers differed significantly from non-smokers at equal probing depths, which was characterized by colonization of novel minority microbes and a shift in abundant members of the microbiome to resemble periodontally diseased communities enriched with pathogenic bacteria. Temporal analysis showed that microbiome in shallow sites were less stable than deeper sites, but temporal stability of the microbiome was not significantly affected by smoking status or scaling and root planing. We identified 7 taxa-Olsenella sp., Streptococcus cristatus, Streptococcus pneumoniae, Streptococcus parasanguinis, Prevotella sp., Alloprevotella sp., and a Bacteroidales sp. that were significantly associated with progression of periodontal disease. Taken together, these results suggest that subgingival dysbiosis in smokers precedes clinical signs of periodontal disease, and support the hypothesis that smoking accelerates subgingival dysbiosis to facilitate periodontal disease progression.

The subgingival microbiome in patients with established rheumatoid arthritis.

OBJECTIVES: To profile and compare the subgingival microbiome of RA patients with OA controls. METHODS: RA (n = 260) and OA (n = 296) patients underwent full-mouth examination and subgingival samples were collected. Bacterial DNA was profiled using 16 S rRNA Illumina sequencing. Following data filtering and normalization, hierarchical clustering analysis was used to group samples. Multivariable regression was used to examine associations of patient factors with membership in the two largest clusters. Differential abundance between RA and OA was examined using voom method and linear modelling with empirical Bayes moderation (Linear Models for Microarray Analysis, limma), accounting for the effects of periodontitis, race, marital status and smoking. RESULTS: Alpha diversity indices were similar in RA and OA after accounting for periodontitis. After filtering, 286 taxa were available for analysis. Samples grouped into one of seven clusters with membership sizes of 324, 223, 3, 2, 2, 1 and 1 patients, respectively. RA-OA status was not associated with cluster membership. Factors associated with cluster 1 (vs 2) membership included periodontitis, smoking, marital status and Caucasian race. Accounting for periodontitis, 10 taxa (3.5% of those examined) were in lower abundance in RA than OA. There were no associations between lower abundance taxa or other select taxa examined with RA autoantibody concentrations. CONCLUSION: Leveraging data from a large case-control study and accounting for multiple factors known to influence oral health status, results from this study failed to identify a subgingival microbial fingerprint that could reliably discriminate RA from OA patients.

Non-antibacterial tetracycline formulations: host-modulators in the treatment of periodontitis and relevant systemic diseases.

Traditionally, the dental profession has primarily treated periodontitis using a mechanical/surgical, rather than a pharmaceutical, approach. However, based on experiments several decades ago which demonstrated that tetracyclines, unexpectedly, inhibit collagen- and bone-destructive mammalian-derived enzymes (e.g. the collagenases), and through non-antibiotic mechanisms, the concept of host-modulation therapy (HMT) was developed. Accordingly, two drug-development strategies evolved: (i) the development of non-antimicrobial formulations of doxycycline; and (ii) the chemical modification of tetracyclines to eliminate their antibiotic activity but retain (or even enhance) their anti-collagenase properties. Regarding the latter, these chemically modified tetracyclines (CMTs) showed efficacy in vitro, in animal models of periodontal (and relevant systemic) disease, and in preliminary clinical trials on patients with Kaposi's sarcoma (however, at the high doses used, photosensitivity was a significant side-effect). In the first strategy, subantimicrobial-dose doxycycline (SDD) demonstrated safety and efficacy in human clinical trials and was approved by the U S Food and Drug Administration (U S FDA) and in other countries for the treatment of periodontitis (20 mg, twice daily, i.e. once every 12 hours) adjunctive to scaling and root planing, and for chronic inflammatory skin diseases (40-mg sustained-release 'beads'). SDD also showed efficacy in patients with systemic diseases relevant to periodontitis, including diabetes mellitus and arthritis, and in postmenopausal women with local and systemic bone loss. Importantly, long-term administration of SDD, of up to 2 years, in clinical trials did not produce antibiotic side-effects. SDD (and in the future, new HMTs, such as low-dose CMT-3, resolvins and chemically modified curcumins) may shift the paradigm of periodontal therapy from a predominantly surgical approach to the greater use of medicinal/pharmacologic strategies, ultimately to benefit larger numbers of patients.

Periodontitis and Porphyromonas gingivalis in patients with rheumatoid arthritis.

OBJECTIVE: To examine the degree to which shared risk factors explain the relationship of periodontitis (PD) to rheumatoid arthritis (RA) and to determine the associations of PD and Porphyromonas gingivalis with pathologic and clinical features of RA. METHODS: Patients with RA (n = 287) and patients with osteoarthritis as disease controls (n = 330) underwent a standardized periodontal examination. The HLA-DRB1 status of all participants was imputed using single-nucleotide polymorphisms from the extended major histocompatibility complex. Circulating anti-P gingivalis antibodies were measured using an enzyme-linked immunosorbent assay, and subgingival plaque was assessed for the presence of P gingivalis using polymerase chain reaction (PCR). Associations of PD with RA were examined using multivariable regression. RESULTS: Presence of PD was more common in patients with RA and patients with anti-citrullinated protein antibody (ACPA)-positive RA (n = 240; determined using the anti-cyclic citrullinated peptide 2 [anti-CCP-2] test) than in controls (35% and 37%, respectively, versus 26%; P = 0.022 and P = 0.006, respectively). There were no differences between RA patients and controls in the levels of anti-P gingivalis or the frequency of P gingivalis positivity by PCR. The anti-P gingivalis findings showed a weak, but statistically significant, association with the findings for both anti-CCP-2 (r = 0.14, P = 0.022) and rheumatoid factor (RF) (r = 0.19, P = 0.001). Presence of PD was associated with increased swollen joint counts (P = 0.004), greater disease activity according to the 28-joint Disease Activity Score using C-reactive protein level (P = 0.045), and higher total Sharp scores of radiographic damage (P = 0.015), as well as with the presence and levels of anti-CCP-2 (P = 0.011) and RF (P < 0.001). The expression levels of select ACPAs (including antibodies to citrullinated filaggrin) were higher in patients with subgingival P gingivalis and in those with higher levels of anti-P gingivalis antibodies, irrespective of smoking status. Associations of PD with established seropositive RA were independent of all covariates examined, including evidence of P gingivalis infection. CONCLUSION: Both PD and P gingivalis appear to shape the autoreactivity of RA. In addition, these results demonstrate an independent relationship between PD and established seropositive RA.

Non-antibacterial tetracycline formulations: clinical applications in dentistry and medicine.

In 1983, it was first reported that tetracyclines (TCs) can modulate the host response, including (but not limited to) inhibition of pathologic matrix metalloproteinase (MMP) activity, and by mechanisms unrelated to the antibacterial properties of these drugs. Soon thereafter, strategies were developed to generate non-antibacterial formulations (subantimicrobial-dose doxycycline; SDD) and compositions (chemically modified tetracyclines; CMTs) of TCs as host-modulating drugs to treat periodontal and other inflammatory diseases. This review focuses on the history and rationale for the development of: (a) SDD which led to two government-approved medications, one for periodontitis and the other for acne/rosacea and (b) CMTs, which led to the identification of the active site of the drugs responsible for MMP inhibition and to studies demonstrating evidence of efficacy of the most potent of these, CMT-3, as an anti-angiogenesis agent in patients with the cancer, Kaposi's sarcoma, and as a potential treatment for a fatal lung disease (acute respiratory distress syndrome; ARDS). In addition, this review discusses a number of clinical studies, some up to 2 years' duration, demonstrating evidence of safety and efficacy of SDD formulations in humans with oral inflammatory diseases (periodontitis, pemphigoid) as well as medical diseases, including rheumatoid arthritis, post-menopausal osteopenia, type II diabetes, cardiovascular diseases, and a rare and fatal lung disease, lymphangioleiomyomatosis.

A comparative evaluation of the sealing ability of 2 root-end filling materials: an in vitro leakage study using Enterococcus faecalis.

OBJECTIVE: The purpose of this study was to evaluate the sealing ability of EndoSequence Bioceramic Root-end Repair (BCRR) material when compared with white mineral trioxide aggregate (WMTA). STUDY DESIGN: Forty single-rooted teeth were instrumented, obturated with gutta-percha, root-end resected, and retrofilled with 2 different materials: white ProRoot MTA (WMTA) (n = 15) and BCRR (n = 15). Unfilled specimens (n = 10) received no retrofill and were used as controls. All groups received E. faecalis in a created reservoir coronal to the root filling and the presence of microleakage was evaluated by counting the colony-forming units from each specimen. The results were analyzed with 1-way analysis of variance. RESULTS: There was no significant difference in leakage between the 2 experimental groups, but there was a significant difference with the control (P ≤ .05). CONCLUSIONS: This study suggests that BCRR is equivalent in sealing ability to WMTA when used as root-end filling material in vitro.

Microbiologic changes following administration of locally delivered doxycycline in smokers: a 15-month follow-up.

BACKGROUND: Additional clinical benefits have been reported with the use of locally delivered antimicrobials in the treatment of smokers; however, long-term changes in the subgingival microbiota following the use of these drugs in smokers have not been documented. The aim of this study was to evaluate long-term microbiologic changes following locally delivered doxycycline in the treatment of smokers. METHODS: Sixteen smokers with chronic periodontitis presenting a minimum of four pockets (probing depth>or=5 mm) were selected. Patients were assigned randomly to receive scaling and root planing (SRP) or SRP and local doxycycline (SRP-D). Patients were treated at baseline and 12 months. Subgingival plaque samples were collected at baseline; 3, 6, and 12 months; and 45 and 90 days following retreatment. Polymerase chain reaction and DNA-DNA hybridization analyses were performed to detect the presence of selected periodontal pathogens. RESULTS: The reduction in the number of sites positive for Porphyromonas gingivalis and Tannerella forsythia (previously T. forsythensis) was statistically significant for SRP-D at 3 months (68% and 41.3%, respectively) and for SRP at 6 months (75% and 52%, respectively) following treatment. The SRP group showed a greater frequency of P. gingivalis than the SRP-D group at 3 months (58% and 25%, respectively). There also was a greater reduction in the frequency of P. gingivalis at 3 months following retreatment with SRP-D compared to SRP (47% and 8%, respectively). CONCLUSION: In smokers, adjunctive local doxycycline resulted in a greater reduction in the frequency of P. gingivalis following initial and supportive therapy compared to conventional treatment.

Subantimicrobial dose doxycycline effects on osteopenic bone loss: microbiologic results.

BACKGROUND: Based on microbiologic concerns, the safety of a 24-month regimen of subantimicrobial dose doxycycline (SDD; 20 mg twice a day) was evaluated in postmenopausal osteopenic women with periodontitis in a double-blind, placebo-controlled, randomized clinical trial. METHODS: Subgingival samples were collected from two sites (probing depth > or = 5 mm) in each of 128 subjects at baseline, with the same sites resampled at the conclusion of the 2-year period. The samples were enumerated on selective and non-selective media and on doxycycline (4 microg/ml) medium. Up to five different colonial morphologies were subcultured from the doxycycline medium, identified to species, and susceptibilities determined to doxycycline and five other antibiotics. Data were analyzed for microbial differences in total colony forming units (CFU), periodontal and opportunistic pathogens, and changes in species and in susceptibilities of isolates recovered on doxycycline medium. RESULTS: There was no significant evidence that changes in total anaerobic counts over the treatment period (P = 0.96) differed between treatment groups. Likewise, periodontal pathogens, opportunistic pathogens, or normal flora did not differ descriptively between groups. Although there was a significant increase (P <0.001) in the total CFU recovered from the 4 microg/ml doxycycline plates at 24 months for SDD versus placebo, the percentage that was clinically resistant to doxycycline (minimal inhibitory concentration [MIC] > or = 16 microg/ml) decreased over the 24-month period in both groups and did not differ between the treatment groups (SDD: 79% to 76%; placebo: 83% to 70%; P = 0.2). There were no significant differences (P >0.28 for each) in the change in cross-resistance between the groups for doxycycline and the other five antibiotics. CONCLUSIONS: No antimicrobial effect on the subgingival flora was detected following treatment with SDD for 24 months, relative to baseline or to placebo. The increase in initial resistance (at 4 microg/ml) did not translate into a significant increase in the percent resistant to doxycycline (MIC > or = 16 microg/ml) for patients in the SDD group.

Current Status of Systemic Antibiotic Usage in Destructive Periodontal Disease.

The recognition that periodontal diseases are primarily caused by specific microorganisms has led researchers to explore the possibility that antibiotics may enhance the effect of mechanical debridement procedures such as scaling and surgery. For some selected periodontal diseases, this has proven to be true. This paper will review systemically-administered antibiotics and the clinical studies and case reports supporting their use. In periodontal therapy, the tetracyclines are the most commonly-used antibiotics in the United States. Tetracycline hydrochloride, minocycline, and doxycycline have been shown to inhibit in vitro most putative periodontal pathogens. Several studies support the use of tetracyclines in the treatment of localized juvenile periodontitis. Penicillins such as amoxicillin are effective in vitro against most periodontal pathogens but have limited efficacy due to the presence of beta-lactamases in gingival fluid. Amoxicillin/ clavulanate potassium (Au) has proven effective in treating adult refractory periodontitis characterized by a Gram-positive flora. Metronidazole is an effective adjunct in adult periodontitis associated with high numbers of "black-pigmented Bacteroides" and spirochetes. A combination of metronidazole and amoxicillin produces a synergistic effect against A. actinomycetemcomitans and has been shown to be effective at eliminating this organism. Clindamycin is an effective adjunct in the treatment of adult refractory periodontitis associated with a predominantly Gram-negative flora. The use of macrolides, quinolones, and combinations of antibiotics is discussed. Clinical studies do not support the use of systemically-administered antibiotics in routine adult periodontitis. Clinical studies do, however, support the use of antibiotics in the treatment of specific periodontal diseases. J Periodontol 1993; 64:760-771.

Efficacy of Clindamycin Hydrochloride in Refractory Periodontitis: 12-Month Results.

The purpose of this study was to evaluate the use of Clindamycin hydrochloride as an adjunct to conventional periodontal therapy in the treatment of patients who had previously been unsuccessfully treated with scaling, periodontal surgery and the use of tetracycline. Thirteen patients with a history of "refractory" periodontitis were thoroughly scaled and monitored by repeated attachment level measurements for the presence of active destructive periodontitis. Disease activity was defined as a 3-mm loss in attachment from baseline measurements or the occurrence of a periodontal abscess. When active disease was detected, each patient was scaled again and placed on Clindamycin hydrochloride 150 mg qid for 7 days. Following the adjunctive use of Clindamycin in combination with scaling, the incidence of gingival sites demonstrating active disease in the group of 13 patients decreased from an annual rate of 10.7 to 0.5%. Each patient demonstrated a decreased incidence of active sites per unit of time. Clinical parameters such as probing depth, gingival redness, bleeding on probing and suppuration showed dramatic improvement at 12 months after Clindamycin therapy. The percentage of pockets with probing depths greater than 6 mm, 4 to 6 mm and 1 to 3 mm changed from 11 to 2%, 38 to 24% and 51 to 74% respectively, following Clindamycin therapy as compared to scaling alone. The percentage of sites bleeding on probing decreased from 33% after scaling alone to 8% following Clindamycin and scaling. Gingival redness decreased from 36 to 1% of sites. Suppuration also decreased from 8% of buccal or lingual surfaces after scaling alone to 1% of surfaces following scaling and Clindamycin. Clindamycin treatment resulted in a significant change in the microbial flora as demonstrated by darkfield microscopy. Prior to Clindamycin therapy, active sites averaged 35% spirochetes, 23% motile rods, 38% nonmotile rods and 4% coccoid cells. At 12 months postclindamycin treatment, spirochetes averaged 8%, motile rods 6%, nonmotile rods 57% and coccoid cells 28% of the organisms present. Our findings indicate that Clindamycin is useful as a treatment modality in cases which do not respond to conventional treatment methods. Due to possible adverse side effects, Clindamycin should be reserved for those patients for whom treatment modalities of lesser risk have been unsuccessful and in whom the subgingival flora is susceptible to this agent.