Patient-specific establishment of bacterial composition within the peri-implant microbiota during the earliest weeks after implant uncovering.

BACKGROUND AND OBJECTIVE: Dysbiosis, a loss of balance in the microbiota, is a potential factor of peri-implantitis. However, compositional change of the peri-implant microbiota soon after implant uncovering is still unknown. In this study, bacterial composition in the peri-implant sulcus was examined to understand the establishment of bacterial composition within the peri-implant microbiota during the earliest weeks after implant uncovering. METHODS: Microbiota samples were collected at weeks 1, 2, 4, and 6 after stage-two surgery. Bacterial DNA was isolated from the samples, and a 16S rRNA gene library was constructed. Sequence reads were obtained using a high-throughput sequencing platform and were taxonomically assigned at the phylum and genus levels. RESULTS: Alpha diversity indices, which did not include taxonomic information, were at similar levels throughout the four time points. At 1 and 2 weeks, the bacterial composition was similar among patients with the predominance of Firmicutes and Proteobacteria. However, the composition was diverse at 4 and 6 weeks and significantly dissimilar to the composition at 1 week. CONCLUSIONS: At 1 week, the peri-implant microbiota was already formed with alpha diversity as high as that at the later time points. However, the bacterial composition was not highly dissimilar among patients at 1 week. The composition changed over the passage of several weeks and was specific for each patient.

Clinical Evaluation of Short (6 mm) and Longer Implants Placed Side by Side in Posterior Partially Edentulous Area: A 3-Year Observational Study.

Background: Short implants have been proposed as an alternative solution for the rehabilitation of atrophic posterior region. Purpose: To compare the clinical outcomes between 6 mm short implants and conventional implants placed under similar conditions of bone quality and occlusal loading. Materials and Methods: Nine patients received atone 6 mm implant and one standard-length (8 mm length or longer) implants in a total of 10 partially edentulous areas. Implants were left submerged for 3-6 months healing period and the screw-retained splinted prostheses were delivered. When the provisional or final restoration was placed, and at each year after loading, standardized intraoral radiograph was taken for themarginal bone level (MBL) changes around the implants. Subsequently, the patients were recalled for the clinical examination evaluating the implant survival, sulcus bleeding index, suppuration, and the incidence of prosthetic complications at every 6 months after the definitive crown delivery. The observation period was continued to 3 years (mean follow-up was 3.4 ± 0.3 years) after functional loading. Results: Nine patients (10 short implants and 10 standard length implants) were selected in this study. Cumulative survival rates of the short implants and standard-length implants were 100% in both groups, and no biological and prosthetic complication were found in 3 years observation period. Cortical bone thickness of implant insertion sites was 1.39 ± 0.45 mm at short implants and 1.38 ± 0.69 mm at standard-length implants, and trabecular bone computed tomography values of implant insertion sites was 424.1 ± 290.1 at short implants and 410.9 ± 267.9 at standard-length implants. The MBL changes were -0.30 ± 0.71 mm at short implants and -0.19 ± 0.78 mm at standard-length implants at 3 years follow-up visit. No significant difference was found in the average of MBL changes between implant length. Conclusions: Within the limits of this study, it can be concluded that 6 mm short implants in a posterior edentulous region showed excellent results compared with conventional implants.

Peri-implantitis management by resective surgery combined with implantoplasty and Er:YAG laser irradiation, accompanied by free gingival graft: a case report.

The optimal method for decontamination of implant surfaces for peri-implantitis treatment remains controversial. In recent years, erbium-doped yttrium aluminum garnet (Er:YAG) laser irradiation and implantoplasty (IP) (i.e. mechanical modification of the implant) have been reported to be effective in decontaminating implant surfaces during the surgical treatment. Also, a lack of adequate keratinized mucosa (KM) around the implant is known to be associated with more plaque accumulation, tissue inflammation, attachment loss, and mucosal recession, increasing the risk of peri-implantitis. Therefore, free gingival graft (FGG) has been recommended for gaining adequate KM around the implant. However, the necessity of acquiring KM for the treatment of peri-implantitis using FGG remains unclear. In this report, we applied the apically positioned flap (APF) as resective surgery for peri-implantitis treatment in conjunction with IP and Er:YAG laser irradiation to polish/clean the implant surface. Furthermore, FGG was conducted simultaneously to create additional KM, which increased the tissue stability and contributed to the positive results. The two patients were 64 and 63 years old with a history of periodontitis. The removal of granulation tissue and debridement of contaminated implant surfaces were performed with Er:YAG laser irradiation post flap elevation and then modified smooth surfaces mechanically using IP. Er:YAG laser irradiation was also utilized to remove the titanium particles. In addition, we performed FGG to increase the width of KM as a vestibuloplasty. Peri-implant tissue inflammation and progressive bone resorption were not observed, and both patients maintained good oral hygiene conditions until the 1-year follow-up appointment. Bacterial analysis via high-throughput sequencing revealed proportional decreases in bacteria associated with periodontitis (Porphyromonas, Treponema, and Fusobacterium). To the best of our knowledge, this study is the first to describe peri-implantitis management and bacterial change before and after procedures by resective surgery combined with IP and Er:YAG laser irradiation for peri-implantitis treatment, accompanied by FGG for increasing KM around the implants.

Autologous micrografts from the palatal mucosa for bone regeneration in calvarial defects in rats: a radiological and histological analysis.

BACKGROUND: The application of dental implants is often restricted by bone volume. In such cases, bone grafts are required, although bone graft materials have some disadvantages. Therefore, other effective approaches are needed. Our previous study showed that the autologous micrograft, a dissociated cell suspension made out of palatal connective tissue grafts, promoted bone-marrow cell proliferation and differentiation under osteogenic conditions. In this study, we aimed to evaluate the effects of dissociated soft-tissue suspensions relevant to bone regeneration in animal model. MATERIAL AND METHODS: Twelve-week-old male Wistar rats were used in the study. Defects were created in rat calvaria, and were filled with hydrogel containing either dissociated soft-tissue suspension (test) or sucrose (control). The new bone formation was evaluated at 1 and 2 weeks after surgery (n = 16) by radiological and histological analysis. RESULTS: The conducted radiological analysis showed that the new bone volume was significantly greater in the dissociated soft-tissue suspension group. This finding was further confirmed by the conducted histological analysis. CONCLUSIONS: The dissociated mucosa tissue suspension enhanced bone regeneration in vivo; thus, it is a promising potential method to aid the successful application for bone augmentation in the implant practice.

Staged Sinus Floor Elevation Using Novel Low-Crystalline Carbonate Apatite Granules: Prospective Results after 3-Year Functional Loading.

The aim of this study was to evaluate clinical outcomes of staged sinus floor elevation (SFE) using novel low-crystalline carbonate apatite (CO3Ap) granules. Patients who needed SFE for implant placement were recruited into this clinical trial. A staged procedure (lateral window technique using CO3Ap granules, followed by implant placement after 7 ± 2 months) was employed in 13 patients. Bone-height increase and insertion torque values (ITVs) were assessed along with histological evaluation. The survival and success rates of 3-year functioning implants were also evaluated. Mean of bone-height increase after SFE using CO3Ap granules was 7.2 ± 2.5 mm and this increase allowed implant placement in all cases (17 implants). Mean of ITV was 25.1 ± 13.2 Ncm and primary stability was achieved successfully in all cases. Histological analyses revealed mature new bone formation (36.8 ± 17.3%) and residual CO3Ap granules (16.2 ± 10.1%) in the compartment after SFE. The survival and success rates after 3-year functional loading were 100% and no complications were found. These results clearly indicate the clinical usefulness of CO3Ap granules for SFE.

Discrimination of Bacterial Community Structures among Healthy, Gingivitis, and Periodontitis Statuses through Integrated Metatranscriptomic and Network Analyses.

Periodontal disease is an inflammatory condition caused by polymicrobial infection. The inflammation is initiated at the gingiva (gingivitis) and then extends to the alveolar bone, leading to tooth loss (periodontitis). Previous studies have shown differences in bacterial composition between periodontal healthy and diseased sites. However, bacterial metabolic activities during the health-to-periodontitis microbiome shift are still inadequately understood. This study was performed to investigate the bacterial characteristics of healthy, gingivitis, and periodontitis statuses through metatranscriptomic analysis. Subgingival plaque samples of healthy, gingivitis, and periodontitis sites in the same oral cavity were collected from 21 patients. Bacterial compositions were then determined based on 16S rRNA reads; taxonomic and functional profiles derived from genes based on mRNA reads were estimated. The results showed clear differences in bacterial compositions and functional profiles between healthy and periodontitis sites. Co-occurrence networks were constructed for each group by connecting two bacterial species if their mRNA abundances were positively correlated. The clustering coefficient values were 0.536 for healthy, 0.600 for gingivitis, and 0.371 for periodontitis sites; thus, network complexity increased during gingivitis development, whereas it decreased during progression to periodontitis. Taxa, including Eubacterium nodatum, Eubacterium saphenum, Filifactor alocis, and Fretibacterium fastidiosum, showed greater transcriptional activities than those of red complex bacteria, in conjunction with disease progression. These taxa were associated with periodontal disease progression, and the health-to-periodontitis microbiome shift was accompanied by alterations in bacterial network structure and complexity. IMPORTANCE The characteristics of the periodontal microbiome influence clinical periodontal status. Gingivitis involves reversible gingival inflammation without alveolar bone resorption. In contrast, periodontitis is an irreversible disease characterized by inflammatory destruction in both soft and hard tissues. An imbalance of the microbiome is present in both gingivitis and periodontitis. However, differences in microbiomes and their functional activities in the healthy, gingivitis, and periodontitis statuses are still inadequately understood. Furthermore, some inflamed gingival statuses do not consistently cause attachment loss. In this study, metatranscriptomic analyses were used to investigate the specific bacterial composition and gene expression patterns of the microbiomes of the healthy, gingivitis, and periodontitis statuses. In addition, co-occurrence network analysis revealed that the gingivitis site included features of networks observed in both the healthy and periodontitis sites. These results provide transcriptomic evidence to support gingivitis as an intermediate state between the healthy and periodontitis statuses.

Effects of implant thread design on primary stability-a comparison between single- and double-threaded implants in an artificial bone model.

BACKGROUND: Primary implant stability is essential for osseointegration. To increase stability without changing the implant size, the thread length must be extended by reducing pitch, using a double-threaded implant, or reducing pitch/lead and lead angle to half that of a single-threaded implant. MATERIALS AND METHODS: We tested the stabilities of these configurations using artificial bone. A 1.2-mm pitch, single-threaded implant (12S) was the control. We tested a 0.6-mm pitch/1.2-mm-lead double-threaded implant (06D) and a 0.6-mm pitch/lead single-threaded implant (06S). We compared stabilities by measuring insertion torque, removal torque, and the implant stability quotient (ISQ). Damage to bone tissue caused by the implants was evaluated using microscopy and morphometric analysis. RESULTS: We show that 06D and 06S significantly improved stability compared with the 12S reference. The stability of 06S was significantly greater compared with that of 06D, except for ISQ. The three implants were associated with bone tissue damage characterized by debris and voids surrounding the implant/bone interface. The 06D caused the most tissue damage, followed by 06S and then 12S. CONCLUSION: These findings indicate that primary stability was significantly improved by changing the implant size, extending the thread length with reduced pitch/lead, and reducing the lead angle to half that of a single-threaded implant compared with a double-threaded implant.

Phylogenetic diversity in fim and mfa gene clusters between Porphyromonas gingivalis and Porphyromonas gulae, as a potential cause of host specificity.

BACKGROUND: Periodontopathic bacteria Porphyromonas gingivalis in humans and Porphyromonas gulae in animals are phylogenetically close and commonly have FimA and Mfa1 fimbriae. However, little is known about how fimA and mfa1 are phylogenetically different between P. gingivalis and P. gulae. Here, we examined phylogenetic diversity in their fim and mfa gene clusters. METHODS: Twenty P. gulae strains were isolated from the periodontal pocket of 20 dogs. For their genomic information, along with 64 P. gingivalis and 11 P. gulae genomes, phylogenetic relationship between the genotypes of fimA and mfa1 was examined. Variability of amino acid sequences was examined in the three-dimensional structure of FimA. The distance between strains was calculated for fim and mfa genes. RESULTS: Some fimA genotypes in P. gulae were close to particular types in P. gingivalis. Two types of mfa1 were classified as 70-kDa and 53-kDa protein-coding mfa1. The variable amino acid positions were primarily at the outer part of FimA. The genes encoding the structural proteins and the main component were similarly distant from the reference strain in P. gingivalis, but not in P. gulae. CONCLUSIONS: The differences in the gene clusters between P. gingivalis and P. gulae may result in their host specificity.

Discriminating Microbial Community Structure Between Peri-Implantitis and Periodontitis With Integrated Metagenomic, Metatranscriptomic, and Network Analysis.

Peri-implantitis and periodontitis are both polymicrobial diseases induced by subgingival plaque accumulation, with some differing clinical features. Studies on the microbial and gene transcription activity of peri-implantitis microbiota are limited. This study aimed to verify the hypothesis that disease-specific microbial and gene transcription activity lead to disease-specific clinical features, using an integrated metagenomic, metatranscriptomic, and network analysis. Metagenomic data in peri-implantitis and periodontitis were obtained from the same 21 subjects and metatranscriptomic data from 12 subjects were obtained from a database. The microbial co-occurrence network based on metagenomic analysis had more diverse species taxa and correlations than the network based on the metatranscriptomic analysis. Solobacterium moorei and Prevotella denticola had high activity and were core species taxa specific to peri-implantitis in the co-occurrence network. Moreover, the activity of plasmin receptor/glyceraldehyde-3-phosphate dehydrogenase genes was higher in peri-implantitis. These activity differences may increase complexity in the peri-implantitis microbiome and distinguish clinical symptoms of the two diseases. These findings should help in exploring a novel biomarker that assist in the diagnosis and preventive treatment design of peri-implantitis.

The Effect of Dissociated Soft Tissue on Osteogenesis: A Preliminary In Vitro Study.

PURPOSE: The purpose of this in vitro study was to examine the effect of dissociated soft tissue on bone marrow cell proliferation and differentiation under osteogenic conditions. MATERIALS AND METHODS: Rat bone marrow cells were cultured to assess the stimulation of cell proliferation and differentiation. Harvested palatal mucosa was dissociated using a device (Rigenera; Human Brain Wave) and the dissociated soft tissue was cultured with rat bone marrow cells. Cell proliferation, differentiation, and mineralized nodule formation were assessed after 2 or 5 days of culturing. Bone marrow cell proliferation was assessed by quantifying the absorbance of a water-soluble tetrazolium salt using a cell proliferation assay kit. Bone marrow cell differentiation was assessed by alkaline phosphatase staining and real-time polymerase chain reaction. Mineralized nodule formation was assessed by Alizarin red staining. RESULTS: At day 2, cell proliferation, osteoblast specific gene expression, and mineralized nodule formation were significantly higher in the experimental group than in the control group. Alkaline phosphatase staining was also higher in the experimental group on day 2. Mineralized nodule formation area and osteoblast specific gene expression were also statistically higher in the experimental group on day 5. CONCLUSION: This study demonstrates that dissociated soft tissue elevates bone marrow cell proliferation and differentiation under osteogenic conditions.

Effects of acidic sodium fluoride-treated, commercially pure titanium on periodontal pathogens and rat bone marrow cells.

The purpose of the present study was to clarify the potentiality of acidic fluoride solution in treating peri-implantitis. We examined bactericidal activity of fluoride solution against periodontal pathogens; and evaluated the effects of fluoride on titanium, and the effects on cell proliferation and differentiation of rat bone marrow cells on the fluoride-treated titanium. Aggregatibacter actinomycetemcomitans and Porphyromonas gingivalis were used to determine minimal inhibitory concentration (MIC) and shorttime exposure. The cells were seeded on the titanium surface with or without fluoride treatment. Then, cellular proliferation, differentiation and mineral deposition were analyzed. The MIC values for A. actinomycetemcomitans and P. gingivalis were 225 and 900 ppm F(-), respectively. In short-time exposure test, both bacterial strains exhibited a significant decrease in a concentrationdependent manner. Cell proliferation and mineral deposition were significantly increased on the fluoride-treated surface. Within the limitation of this study, acidic sodium fluoride solution has the potentiality in treating peri-implantitis.