Anton Friedmann

Filifactor alocis - involvement in periodontal biofilms

BackgroundBacteria in periodontal pockets develop complex sessile communities that attach to the tooth surface. These highly dynamic microfloral environments challenge both clinicians and researchers alike. The exploration of structural organisation and bacterial interactions within these biofilms is critically important for a thorough understanding of periodontal disease. In recent years, Filifactor alocis, a fastidious, Gram-positive, obligately anaerobic rod was repeatedly identified in periodontal lesions using DNA-based methods. It has been suggested to be a marker for periodontal deterioration. The present study investigated the epidemiology of F. alocis in periodontal pockets and analysed the spatial arrangement and architectural role of the organism in in vivo grown subgingival biofilms.ResultsA species-specific oligonucleotide probe, FIAL, was designed and evaluated. A total of 490 subgingival plaque samples were submitted to PCR and subsequent dot blot hybridization to compare the prevalence of F. alocis in patients suffering from generalized aggressive periodontitis (GAP), chronic periodontitis (CP), and control subjects resistant to periodontitis. Moreover, a specially designed carrier system was used to collect in vivo grown subgingival biofilms from GAP patients. Subsequent topographic analysis was performed using fluorescence in situ hybridization.While the majority of patients suffering from GAP or CP harboured F. alocis, it was rarely detected in the control group. In the examined carrier-borne biofilms the organism predominantly colonized apical parts of the pocket in close proximity to the soft tissues and was involved in numerous structures that constitute characteristic architectural features of subgingival periodontal biofilms.ConclusionsF. alocis is likely to make a relevant contribution to the pathogenetic structure of biofilms accounting for periodontal inflammation and can be considered an excellent marker organism for periodontal disease.

Ridge augmentation and maxillary sinus grafting with a biphasic calcium phosphate: histologic and histomorphometric observations

OBJECTIVES This retrospective study reports on histologic and histomorphometric observations performed on human biopsies harvested from sites augmented exclusively by biphasic calcium phosphate [BCP: hydroxyapatite (HA)/ tricalcium phosphate (TCP) 60/40] and healed for a minimum of 6 months. MATERIALS AND METHODS Five patients benefited from three augmentation regimens (i.e.: one-stage lateral augmentation; two-stage lateral augmentation; and two-stage sinus grafting). In all patients, a degradable collagen membrane served as a cell-occlusive barrier. Core biopsies were obtained from lateral as from crestal aspects 6-10 months after augmentation surgeries. For histologic and histomorphometric evaluations, the non-decalcified tissue processing was performed. RESULTS The histological examination of 11 biopsies showed graft particles frequently being bridged by the new bone, and a close contact between the graft particles and newly formed bone was seen in all samples. The mean percentages of newly formed bone, soft tissue compartment, and graft material were 38.8% (+/-5.89%), 41.75% (+/-6.08%), and 19.63% (+/-4.85%), respectively. Regarding bone-to-graft contact values, the percentage of bone coverage of graft particles for all biopsies ranged from 27.83% to 80.17%. The mean percentage of bone coverage was 55.39% (+/-13.03%). CONCLUSIONS Data from the present study demonstrated osteoconductivity scores for the BCP material (HA/TCP 60/40) in patients resembling those previously shown for grafting materials of xenogenic and alloplastic origin.

Are periodontal diseases really silent? A systematic review of their effect on quality of life

AIM Periodontal diseases (PDs) may play an important role in the effect oral health status has on a persons quality of life (QoL). The objective was to investigate the influence of PDs (gingivitis and periodontitis) on oral health-related QoL (OHRQoL) by systematically reviewing the literature. MATERIALS AND METHODS Studies using clinical periodontal examinations and validated survey instruments were included. Among 1134 citations initially identified through electronic and hand searching, 37 were eligible and data were extracted from full texts. A vote counting method was used for synthesis of the results. RESULTS Included studies were published between 2001 and 2014 and revealed considerable heterogeneity in participant selection, clinical assessments, and OHRQoL measures. A significant association between PDs and OHRQoL was reported in 28 studies, of which eight reported increasing impact with greater disease severity or extent. CONCLUSIONS Within the limits of the available literature, OHRQoL was affected by clinically assessed PDs. There was evidence for increased impairment with greater severity and extent of PDs, and the recognition of the association was increased when full mouth recording protocols were applied.

Co-localized or randomly distributed? Pair cross correlation of in vivo grown subgingival biofilm bacteria quantified by digital image analysis.

The polymicrobial nature of periodontal diseases is reflected by the diversity of phylotypes detected in subgingival plaque and the finding that consortia of suspected pathogens rather than single species are associated with disease development. A number of these microorganisms have been demonstrated in vitro to interact and enhance biofilm integration, survival or even pathogenic features. To examine the in vivo relevance of these proposed interactions, we extended the spatial arrangement analysis tool of the software daime (digital image analysis in microbial ecology). This modification enabled the quantitative analysis of microbial co-localization in images of subgingival biofilm species, where the biomass was confined to fractions of the whole-image area, a situation common for medical samples. Selected representatives of the disease-associated red and orange complexes that were previously suggested to interact with each other in vitro (Tannerella forsythia with Fusobacterium nucleatum and Porphyromonas gingivalis with Prevotella intermedia) were chosen for analysis and labeled with specific fluorescent probes via fluorescence in situ hybridization. Pair cross-correlation analysis of in vivo grown biofilms revealed tight clustering of F. nucleatum/periodonticum and T. forsythia at short distances (up to 6 µm) with a pronounced peak at 1.5 µm. While these results confirmed previous in vitro observations for F. nucleatum and T. forsythia, random spatial distribution was detected between P. gingivalis and P. intermedia in the in vivo samples. In conclusion, we successfully employed spatial arrangement analysis on the single cell level in clinically relevant medical samples and demonstrated the utility of this approach for the in vivo validation of in vitro observations by analyzing statistically relevant numbers of different patients. More importantly, the culture-independent nature of this approach enables similar quantitative analyses for “as-yet-uncultured” phylotypes which cannot be characterized in vitro.

Soft tissue expansion with self-filling osmotic tissue expanders before vertical ridge augmentation: a proof of principle study

INTRODUCTION Post-surgical graft exposition and loss of grafted bone are a common complication of vertical bone augmentation. Soft tissue expansion (STE) by implantation of osmotic self-filling tissue expanders before reconstructive surgery is an effective method for generation of soft tissue. The aim of this study was to investigate the feasibility of STE before bone augmentation with regard to clinical and histological outcomes and complications. METHODS Tissue expanders were implanted in patients requiring vertical bone augmentation. Onlay grafting was carried out after 2 months of STE. Implants were placed 4-6 months after augmentation. Vertical bone gain was analysed with cone-beam computed tomography (CBCT). Bone biopsies were investigated with micro-computed tomography (micro-CT). RESULTS Twenty-four sites in 12 patients were treated with STE. Complications of STE were perforation (two sites) and infection (two sites). At augmentation after STE, primary wound closure was easily achieved and the incidence of graft expositions was low (4%). At implant placement, high vertical bone gain of 7.5 ± 2.4 mm was found. Micro-CTs of bone revealed a good ratio of bone volume/tissue volume (mean BV/TV=0.1614 ± 0.0582). All implants were osseointegrated. CONCLUSIONS The combination of STE and subsequent vertical augmentation provided high gain of well-structured bone for further successful implant therapy and was accompanied by minimal complications.

Molecular epidemiology and spatial distribution of Selenomonas spp. in subgingival biofilms.

The aetiology of periodontal disease has been a field of intensive research in the past decades. Along with a variety of other putative pathogens, different members of the genus Selenomonas have repeatedly been associated with both generalized aggressive periodontitis and chronic periodontitis. For the present study, a specific oligonucleotide probe targeting the majority of all oral Selenomonas spp. was designed. Their prevalence was determined, using dot-blot hybridization, in a total of 742 subgingival samples collected from patients with generalized aggressive (n=62) and chronic periodontitis (n=82), and from periodontitis-resistant subjects (n=19). In addition, fluorescence in situ hybridization (FISH) and electron microscopy were performed to analyze the spatial arrangement of Selenomonas in subgingival biofilms collected from patients with generalized aggressive periodontitis. In the samples from patients, Selenomonas spp. showed a lower prevalence in both diseased groups compared with other putative pathogens, and a relatively high prevalence in the periodontitis-resistant group. Consequently, Selenomonas spp. do not seem to be suitable diagnostic marker organisms for periodontal disease. By contrast, FISH and electron microscopic analysis of periodontal carriers revealed that Selenomonas spp. appeared in large numbers in all parts of the collected biofilms and seemed, if present in a site from patients, to make a relevant contribution to their structural organization.

Randomized controlled trial on lateral augmentation using two collagen membranes: morphometric results on mineralized tissue compound

BACKGROUND Guided bone regeneration is considered an effective tool for gaining mineralized tissue either at exposed implant surface or in deficient alveolar ridge areas before implant placement. MATERIAL AND METHODS Customized casts obtained following impression taking at surgery and re-entry allowed for morphometric assessment of alveolar ridge alterations 6 months after one-stage augmentation of bone dehiscences. In a randomized pilot study using biphasic calcium phosphate tests (n=17) received treatment with ribose cross-linked collagen membranes (RCLM), whereas controls (n=20) received non-cross-linked membranes. The primary endpoint was to quantify the effect of membrane type on dimensional changes in bone margins at crestal level of endosseous implants. RESULTS Soft tissue dehiscencies occurred at 70.5% and 55% frequency for tests and controls, respectively. Gain in clinically hard newly mineralized tissue at the crestal level was significantly higher in test group in lateral (1.8 versus 0.7 mm; p=.046) and in vertical dimensions (1.1 versus 0.2 mm; p=.035) compared with controls. Second measurement obtained at the border of reflected flap revealed no significant difference between groups (3.0 versus 2.1 mm; p=0.57) for lateral dimension. CONCLUSIONS Both collagen devices were effective in bone augmentation. RCLMs supported mineralization process and remodelling even in sites showing compromised healing as indicated by morphometric outcome.

Calprotectin levels in gingival crevicular fluid predict disease activity in patients treated for generalized aggressive periodontitis.

BACKGROUND AND OBJECTIVE Clinical parameters such as probing depth and bleeding on probing are commonly used for monitoring after periodontal treatment. However, these parameters have poor prognostic utility. The biomarker calprotectin is used to monitor conditions such as inflammatory bowel disease because of its ability to predict disease activity. Levels of calprotectin in gingival crevicular fluid correlate with periodontal disease severity and treatment outcome. The validity of calprotectin as predictor for future periodontal disease activity has not yet been investigated. MATERIAL AND METHODS Thirty-six subjects with generalized aggressive periodontitis were treated with scaling and root planing (SRP), and with adjunctive antimicrobial medications. Probing depth, clinical attachment level and bleeding on probing were assessed at baseline, and 3 and 6mo after SRP. A gingival crevicular fluid sample was collected from the initially deepest site in each patient 3mo after SRP and analysed for calprotectin levels. Activity was defined as a probing depth increase of >0.5mm between 3 and 6mo at the sample site. The ability of individual parameters to predict activity was analysed by construction of receiver operating characteristic curves. RESULTS Nine active sites were identified. Clinical attachment level, probing depth, bleeding on probing and gingival crevicular fluid volume showed no predictive utility [area under the curve (AUC) <0.6, p>0.05]. However, calprotectin concentration (AUC=0.793, p=0.01) and the total amount/sample of calprotectin (AUC=0.776, p=0.02) significantly predicted activity. Patients with calprotectin levels above calculated cut-off values had significantly more active sites than patients with negative results. CONCLUSION Calprotectin levels were predictors of disease activity at both site and subject levels. The calculated cut-off values provide a dichotomous basis for prospective evaluation of calprotectin as a diagnostic marker for monitoring periodontal treatment.

Improvement of microcirculation and wound healing in vertical ridge augmentation after pre-treatment with self-inflating soft tissue expanders – a randomized study in dogs

OBJECTIVES We investigated the effect of soft tissue expansion (STE) on vertical ridge augmentation with regard to the incidence of wound dehiscences and the impairment of microcirculation in dogs, and the applicability of laser Doppler flowmetry (LDF) to explore the relation between microcirculation and wound healing. MATERIAL AND METHODS Bone defects were created on both mandibular sides in ten beagle dogs by extraction of premolars and removal of bone. Six weeks later, self-filling tissue expanders were implanted in randomly assigned test sites. After 5 weeks of expansion, vertical augmentation was carried out in test and control sites using calvarial onlay grafts side by side with granular biphasic calcium phosphate covered with a resorbable polyethylene glycol membrane. Microcirculation was evaluated with laser Doppler flowmetry (LDF). The incidence of wound dehiscences was evaluated after 2 weeks. The validity of LDF to predict dehiscences was evaluated by construction of receiver operating characteristic (ROC) curves. RESULTS After augmentation, test sites showed significantly better perfusion than control sites without preceding STE (P = 0.012). Three days after surgery, perfusion was still significantly decreased in control sites (P = 0.005), while microcirculation in test sites had returned to pre-surgical levels. After 2 weeks, healing in test sites was good, whereas eight dehiscences were found in control sites (P = 0.002). ROC curves showed that microcirculation levels immediately after augmentation surgery significantly predicted subsequent wound dehiscences (AUC = 0.799, CI 0.642-0.955, P = 0.006). CONCLUSIONS Laser Doppler flowmetry is suitable for evaluation of soft tissue microcirculation after ridge augmentation. STE reduced the impairment of microcirculation caused by vertical ridge augmentation and decreased the incidence of wound dehiscences in the investigated animal model.

Convergent methods assessing bone growth in an experimental model at dental implants in the minipig

Implant dentistry demonstrated its reliability in treating successfully an increased amount of patients with dental implants exhibiting hydrophilic (modSLA) or non-hydrophilic (SLA) surfaces. Objectives of this minipig study consisted in assessing bone regeneration at both types of dental implants in a new experimental model (lateral bone defects) by implementing a convergent analysis approach combining histology and contact radiography. In six adult female minipigs standardized acute alveolar defects were created, then receiving two implants in the mandibles bilaterally (modSLA and SLA, one of each combined with Guided Bone Regeneration). Animals were sacrificed after 28 days of healing. Mid-crestal specimen were analyzed assessing missing BIC to implant shoulder (fBIC); vertical bone growth upon implant surface, bone:tissue ratio and grafting material area occupied by material remnants for GBR sites. Values obtained from both analyses were compared and statistical correlations scrutinized. Although dimensions of mean differences and adjusted means given by radiographic method were lower than histological ones, comparison of different implants yielded similar results. Statistical analyses of correlation and concordance coefficients used to evaluate radiological method of measurement showed high level of concordance (concordance coefficient=0.912 and correlation coefficient=0.939) for fBIC. Similar results were observed for vertical new bone and for remnants of graft. Discrepancies for new bone and for mineralized tissue resulted in concordance coefficient of 0.182 and 0.054. The results indicate that contact X-rays can be used for morphometric assessments regarding defect fill; however, histological staining remains beneficial if greater resolution for distinguishing qualitative differences in the tissues is required.