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Objective The study aims to investigate the effect of reduced keratinized mucosa (KM) and other risk indicators on the severity of peri-implant mucositis in (i) the general population, (ii) in periodontally healthy patients, and (iii) in periodontally healthy patients without a history of periodontitis. Materials and methods Anamnesis and the following clinical parameters were taken: mucosal-index, bleeding on probing, local plaque index, oral hygiene-index, and width of KM. Mucositis severity score was determined for each implant. Multi-level and subgroup analysis was performed on the patient and implant level. Results Six hundred twelve implants in 130 patients were analyzed. Subgroup analysis showed significant associations between KM < 2 mm and the severity score in (ii) periodontally healthy patients ( p  = 0.014) and in (iii) patients without history of periodontitis ( p  = 0.017). Secondary outcome showed higher severity scores for patients with insufficient oral hygiene or without residual teeth ( p  ≤ 0.001), in maxillary implants ( p  = 0.04), and for the number of implants per patient ( p  ≤ 0.001). Conclusion Within the limits of the study, one may conclude that a reduced width of KM is a risk indicator for the severity of peri-implant mucositis in periodontally healthy patients and patients without a history of periodontitis. Clinical relevance The results indicate a band of ≥ 2 mm KM to reduce the severity of peri-implant mucositis in periodontally healthy patients.

Even though metagenomics have revolutionized the characterization of the human microbiome, detailed mechanistic studies are impracticable, as there is a dearth of robust culture collections. We now describe the development and use of a laser-assisted culturomics platform, incorporating the elements of a bioprinter, the culture conditions, the methods to characterize the microorganisms and a biobank. With laser-assisted bioprinting, the microorganisms can be rapidly and precisely transferred from clinical biofilms to highly organized arrays of microbial colonies, which are suitable for co-culturing and molecular analyses. The presented technique has propagated 99 of 100 microbial species and recovered 79% of abundant species from dental plaque in accordance with full 16S rRNA gene profiling of 691,199 sequences. Microscopy, spectroscopy and enzyme assays have been used to guide isolations. Processing of oral biofilms from four individuals has yielded 249 representative isolates, from 14 classes and 124 species in total. Functional profiling with bioprinting has indicated commensals which could potentially contribute to disease development. Isolates from peri-implantitis cover 85.4% of the transcriptionally active clinical biofilms at genus level. Taken together, this work provides the basis for generating on-demand culture collections and biofilms for research and clinical use. Representative microbial isolates and patient-specific biobanks are crucial for microbiome investigation and management. Here, authors develop laser-assisted microbial culturomics, combining high-throughput, precise bioprinting on diverse media with rapid, non-invasive analyses.

Background Peri-implant mucositis and peri-implantitis are highly prevalent biofilm-associated diseases affecting the tissues surrounding dental implants. As antibiotic treatment is ineffective to fully cure biofilm mediated infections, antimicrobial modifications of implants to reduce or prevent bacterial colonization are called for. Preclinical in vivo evaluation of the functionality of new or modified implant materials concerning bacterial colonization and peri-implant health is needed to allow progress in this research field. For this purpose reliable animal models are needed. Methods Custom made endosseous dental implants were installed in female Sprague Dawley rats following a newly established three-step implantation procedure. After healing of the bone and soft tissue, the animals were assigned to two groups. Group A received a continuous antibiotic treatment for 7 weeks, while group B was repeatedly orally inoculated with human-derived strains of Streptococcus oralis , Fusobacterium nucleatum and Porphyromonas gingivalis for six weeks, followed by 1 week without inoculation. At the end of the experiment, implantation sites were clinically assessed and biofilm colonization was quantified via confocal laser scanning microscopy. Biofilm samples were tested for presence of the administered bacteria via PCR analysis. Results The inner part of the custom made implant screw could be identified as a site of reliable biofilm formation in vivo . S. oralis and F. nucleatum were detectable only in the biofilm samples from group B animals. P. gingivalis was not detectable in samples from either group. Quantification of the biofilm volume on the implant material revealed no statistically significant differences between the treatment groups. Clinical inspection of implants in group B animals showed signs of mild to moderate peri-implant mucositis (4 out of 6) whereas the mucosa of group A animals appeared healthy (8/8). The difference in the mucosa health status between the treatment groups was statistically significant ( p  = 0.015). Conclusions We developed a new rodent model for the preclinical evaluation of dental implant materials with a special focus on the early biofilm colonization including human-derived oral bacteria. Reliable biofilm quantification on the implant surface and the symptoms of peri-implant mucositis of the bacterially inoculated animals will serve as a readout for experimental evaluation of biofilm-reducing modifications of implant materials.

Biofilm-associated peri-implant infections pose a major problem in modern medicine. The understanding of biofilm development is hampered by biofilm complexity and the lack of robust clinical models. This study comprehensively characterized the dynamics of early biofilm formation in the transmucosal passage of implant abutments in 12 patients. Biofilm structures and compositions were complex, diverse, subject-specific and dynamic. A total of 371 different bacterial species were detected. 100 phylogenetically diverse unnamed species and 35 taxonomically diverse disease-associated species comprised an average 4.3% and 3.1% of the community, respectively, but reached up to 12.7% and 21.7% in some samples. Oral taxa formed numerous positive associations and clusters and were characterized by a high potential for metabolic interactions. The subspecies diversity was highly patient-specific and species-dependent, with 1427 ASVs identified in total. The unprecedented depth of early biofilm characterization in this study will support the development of individualized preventive and early diagnostic strategies.

Background The prevalence of peri-implantitis ranges between 7 and 38.4% depending on risk indicators such as smoking, diabetes mellitus, lack of periodontal maintenance program, and history or presence of periodontitis. Currently, the possible effect of the type of superstructure on peri-implant health is unclear. This cross-sectional study aims to investigate the influence of the superstructure on the prevalence of peri-implant mucositis, peri-implantitis and peri-implant dysbiosis. Methods During a 32-month recruitment period dental implants were assessed to diagnose healthy peri-implant tissues, mucositis or peri-implantitis. The study included 1097 implants in 196 patients. Out of all peri-implantitis cases 20 randomly chosen submucosal biofilms from implants with fixed denture (FD) originating from 13 patients and 11 biofilms from implants with removable dentures (RD) originating from 3 patients were studied for microbiome analysis. Composition of transcriptionally active biofilms was revealed by RNAseq. Metatranscriptomic profiles were created for thirty-one peri-implant biofilms suffering from peri-implantitis and microbiome changes associated with superstructure types were identified. Results 16.41% of the implants were diagnosed with peri-implantitis, 25.00% of implants with RD and 12.68% of implants with FD, respectively. Multivariate analysis showed a significant positive association on patient ( p  =  < 0.001) and implant level ( p  = 0.03) between the prevalence of peri-implantitis and RD. Eight bacterial species were associated either with FD or RD by linear discriminant analysis effect size method. However, significant intergroup confounders (e.g. smoking) were present. Conclusions Within the limitations of the present work, RDs appear to be a risk indicator for peri-implantitis and seem to facilitate expansion of specific periodontopathogens. Potential ecological and pathological consequences of shift in microbiome from RDs towards higher activity of Fusobacterium nucleatum subspecies animalis and Prevotella intermedia require further investigation.

Peri-implantitis is a severe biofilm-associated infection affecting millions worldwide. This cross-sectional study aimed to identify taxonomic and functional biomarkers that reliably indicate peri-implantitis by utilizing paired data from full length 16S rRNA gene amplicon sequencing (full-16S) and metatranscriptomics (RNAseq) in 48 biofilm samples from 32 patients. Both full-16S and RNAseq analyses revealed significant differences between healthy and peri-implantitis samples, with a shift toward anaerobic Gram-negative bacteria in peri-implantitis. Metatranscriptomics identified enzymatic activities and metabolic pathways associated with peri-implantitis and uncovered complex peri-implant biofilm ecology related to amino acid metabolism. Integrating taxonomic and functional data enhanced predictive accuracy (AUC = 0.85) and revealed diagnostic biomarkers including health-associated Streptococcus and Rothia species and peri-implantitis-associated enzymes (urocanate hydratase, tripeptide aminopeptidase, NADH:ubiquinone reductase, phosphoenolpyruvate carboxykinase and polyribonucleotide nucleotidyltransferase). Thus, biofilm profiling at taxonomic and functional levels provides highly predictive disease biomarkers, laying the foundation for novel diagnostic and personalized treatment approaches for peri-implant disease.

Background This cross-sectional study investigates the potential association between active periodontal disease and high HbA1c levels in type-2-diabetes mellitus subjects under physical training. Methods Women and men with a diagnosis of non-insulin-dependent diabetes mellitus and ongoing physical and an ongoing exercise program were included. Periodontal conditions were assessed according to the CDC-AAP case definitions. Venous blood samples were collected for the quantitative analysis of HbA1c. Associations between the variables were examined with univariate and multivariate regression models. Results Forty-four subjects with a mean age of 63.4 ± 7.0 years were examined. Twenty-nine subjects had no periodontitis, 11 had a moderate and 4 had a severe form of periodontal disease. High fasting serum glucose ( p  < 0.0001), high BMI scores ( p  = 0.001), low diastolic blood pressure ( p  = 0.030) and high probing depth ( p  = 0.036) were significantly associated with high HbA1c levels. Conclusions Within the limitations of this study HbA1c levels are positively associated with high probing pocket depth in patients with non-insulin-dependent diabetes mellitus under physical exercise training. Control and management of active periodontal diseases in non-insulin-dependent patients with diabetes mellitus is reasonable in order to maximize therapeutic outcome of lifestyle interventions.

Background Despite considerable improvements in oral health in recent decades, caries and periodontitis are still widespread, ranking among the most prevalent diseases worldwide and requiring future research. The German National Cohort (NAKO Gesundheitsstudie, NAKO) is a large-scaled, multidisciplinary, nationwide, multi-centre, population-based, prospective cohort study with oral examinations that aims to provide a resource to study risk factors for major diseases. The aim of the present article is to provide the methodological background, to report on the data quality, and to present initial results of the oral examinations. Methods During baseline examinations (2014–2019), a total of 205,184 persons aged 19–74 years has been examined in 18 study centres, including, among others, a dental interview, stimulated saliva sampling, and recording of the numbers of present teeth and prostheses (standard Level 1 program). As part of the Level 2 program that was offered to 20% randomly selected participants, each study centre selected one of three modules, one of them being the Level 2 oral examination. This extended program was carried out in a subgroup of 20,828 participants, including collection of detailed information on the dental and prosthetic status as well as on periodontal, cariological and functional aspects. To ensure reliability and reproducibility, study nurses were trained and calibrated by dental experts. In addition, a reliability study was conducted among 794 Level 1 and 359 Level 2 participants, reporting intra class correlation and kappa coefficients. Results Intra class correlation and kappa coefficients for observer agreement and reliability were consistently above 0.7, indicating good to excellent reliability of all dental measurements. For example, intra class correlation was 0.937 for the number of present teeth (Level 1), 0.740 for mean probing depth (PD) and 0.797 for active mouth opening. An initial inspection of the data showed that the median number of present teeth was 27, of which on average 6.9 teeth were healthy and caries-free. Average mean PD was 1.92 mm. An orthodontic treatment was reported by 35.5% of participants. Discussion Overall, the dental study protocol was feasible and successfully integrated into the NAKO’s overall assessment program. However, rigorous support of the study centres by dental professionals was required to ensure high quality data. In summary, high-quality data collection within the NAKO pave the way for future investigation of potential risk factors for oral diseases and links between oral and systemic diseases and conditions.