The influence of immediate occlusal loading on micro/nano-structure of peri-implant jaw bone in rats.

PURPOSE: The objective of the present study was to ascertain the effect of immediate occlusal loading after implant placement on osseointegration and the micro/nanostructure of the surrounding bone. METHODS: After extraction of a rat maxillary right second molar, an implant was placed immediately with initial fixation (2 N< ). The implants were placed to avoid occlusal loading due to mastication, and in the loaded group, a superstructure was fabricated and subjected to occlusal loading. Bone morphometry, collagen fiber anisotropy, and biological apatite (BAp) crystallite alignment were quantitatively evaluated in both groups after extraction and fixation of the jaw bone at Days 7 and 21 after surgery. RESULTS: Osseointegration was observed in both groups. Bone morphometry showed significant differences in bone volume, trabecular number, trabecular thickness and bone mineral density (BMD) at Days 21 postoperatively (P < 0.05). A significant difference was also found in the trabecular separation at Days 7 postoperatively (P < 0.05). In the evaluation of collagen fiber anisotropy, collagen fiber bundles running differently from the existing bone were observed in both groups. In terms of BAp crystallite alignment, a specific structure was observed in the reconstructed new bone after implantation, and preferential orientation of BAp crystallite alignment was observed in the longitudinal direction of the implants in the Day 21 postoperative loaded group. CONCLUSION: When sufficient initial fixation is achieved at the time of dental implant placement, then the applied masticatory load may contribute to rapidly achieving not only bone volume, but also adequate bone quality after implant placement.

Effects of the application of low-temperature atmospheric plasma on titanium implants on wound healing in peri-implant connective tissue in rats.

PURPOSE: This study aimed to clarify the effects of surface modification of titanium (Ti) implants by low-temperature atmospheric pressure plasma treatment on wound healing and cell attachment for biological sealing in peri-implant soft tissue. METHODS: Hydrophilization to a Ti disk using a handheld low-temperature atmospheric pressure plasma device was evaluated by a contact angle test and compared with an untreated group. In in vivo experiments, plasma-treated pure Ti implants using a handheld plasma device (experimental group: PL) and untreated implants (control group: Cont) were placed into the rat upper molar socket, and samples were harvested at 3, 7 and 14 days after surgery. Histological evaluation was performed to assess biological sealing, collagen- and cell adhesion-related gene expression by reverse transcription quantitative polymerase chain reaction, collagen fiber detection by Picrosirius Red staining, and immunohistochemistry for integrins. RESULTS: In in vivo experiments, increased width of the peri-implant connective tissue (PICT) and suppression of epithelial down growth was observed in PL compared with Cont. In addition, high gene expression of types I and XII collagen at 7 days and acceleration of collagen maturation was recognized in PL. Strong immunoreaction of integrin α2, α5, and ß1 was observed at the implant contact area of PICT in PL. CONCLUSIONS: The handheld low-temperature atmospheric pressure plasma device provided hydrophilicity on the Ti surface and maintained the width of the contact area of PICT to the implant surface as a result of accelerated collagen maturation and fibroblast adhesion, compared to no plasma application.

Subgingival titanium wire implantation induces weak inflammatory responses but does not promote substantial T cell activation.

Titanium is a biocompatible material commonly used for dental treatments. However, the detailed mechanism underlying the weak biological activity of titanium has not been elucidated. We investigated both the inflammatory responses and T cell activation induced by solid titanium in the gingiva in mice. Both titanium and nickel wire implantation promoted neutrophil infiltration into the gingiva on day 2. Nickel, but not titanium, wire implantation enhanced proinflammatory cytokine expression and dendritic cell activity in gingival tissue by day 2. Nickel wire implantation enhanced the activity of T cells in draining lymph nodes on day 5. Moreover, T cell and neutrophil infiltration and elevated proinflammatory cytokine expression in the gingival tissue were still observed on day 5. However, no such augmented biological responses were observed after titanium wire implantation. These findings suggest that, unlike nickel, solid titanium does not induce sufficient inflammatory responses leading to T cell activation in gingival tissue.

Simultaneous prosthodontic treatment for crowns of natural teeth and dental implants using digital technology: A case report.

The aim of this clinical report was to describe the improvement of masticatory disorders with the use of digital technology to simultaneously perform prosthodontic treatment of natural teeth and edentulous areas. Computer-guided implant surgery was performed, and crown prostheses and implant superstructures were fabricated simultaneously using digital technology.

Subset of the periodontal ligament expressed leptin receptor contributes to part of hard tissue-forming cells.

The lineage of periodontal ligament (PDL) stem cells contributes to alveolar bone (AB) and cementum formation, which are essential for tooth-jawbone attachment. Leptin receptor (LepR), a skeletal stem cell marker, is expressed in PDL; however, the stem cell capacity of LepR+ PDL cells remains unclear. We used a Cre/LoxP-based approach and detected LepR-cre-labeled cells in the perivascular around the root apex; their number increased with age. In the juvenile stage, LepR+ PDL cells differentiated into AB-embedded osteocytes rather than cementocytes, but their contribution to both increased with age. The frequency of LepR+ PDL cell-derived lineages in hard tissue was < 20% per total cells at 1-year-old. Similarly, LepR+ PDL cells differentiated into osteocytes following tooth extraction, but their frequency was < 9%. Additionally, both LepR+ and LepR- PDL cells demonstrated spheroid-forming capacity, which is an indicator of self-renewal. These results indicate that both LepR+ and LepR- PDL populations contributed to hard tissue formation. LepR- PDL cells increased the expression of LepR during spheroid formation, suggesting that the LepR- PDL cells may hierarchically sit upstream of LepR+ PDL cells. Collectively, the origin of hard tissue-forming cells in the PDL is heterogeneous, some of which express LepR.

Dimensional changes of buccal bone in the edentulous maxilla with telescopic-retained implant-supported fixed dental prostheses.

STATEMENT OF PROBLEM: The buccal bone in an edentulous maxilla loses thickness over time because of physiological changes. However, the dimensional changes of the buccal bone in an edentulous maxilla with an implant-supported fixed dental prosthesis are unknown. PURPOSE: The purpose of this retrospective clinical study was to evaluate cone beam computed tomography (CBCT) images of the dimensional changes of the buccal bone in edentulous maxillae with complete arch telescopic-retained implant-supported fixed dental prostheses (CTI-FDPs) after 6 years by using a professional retrieval system. MATERIAL AND METHODS: This study included 17 participants with edentulous maxillae who had been provided with CTI-FDP with 121 taper joint implants. A three-dimensional radiographic analysis by using CBCT was performed at implant insertion (0 years) and after 6 years. Vertical and horizontal bone measurement values were evaluated. During horizontal bone thickness measurement, 4 different levels, 0, 2, 4, and 6 mm apical to the implant shoulder, were evaluated as bone value (BV)0mm, BV2mm, BV4mm, and BV6mm, respectively. The BVs were compared with the Wilcoxon signed-rank test and Kruskal-Wallis test (α=.05). In addition, the Spearman rank correlation coefficient was used to identify 0yBV factors that influence the 6yBVs. A nonlinear regression analysis was used to clarify the slopes of 0yBVs and 6yBV0mm. RESULTS: Significant decreases in vertical and horizontal BVs were found between 0 years and 6 years (P<.05). However, no significant difference was observed in bone loss at 6 years at any of the vertical and horizontal measurement points (P≥.05). When 0yBVs related to 6yBV0mm were analyzed, 0yBV0mm and 0yBV2mm showed strong correlations with 6yBV0mm (|r|≥.7). In the regression analysis, a 0yBV0mm of 0.58 mm and 0yBV2mm of 0.78 mm could be critical factors associated with a 6yBV0mm of 0 mm. A 6yBV0mm of 0yBV0mm more than 0.58 mm was significantly higher than a 6yBV0mm of 0yBV0mm less than 0.58 mm (P<.001). Moreover, a 6yBV0mm of 0yBV2mm more than 0.78 mm was significantly higher than a 6yBV0mm of 0yBV2mm less than 0.78 mm (P<.001). CONCLUSIONS: The buccal bone in an edentulous maxilla with fixed implant-supported prostheses lost significant vertical and horizontal bone thicknesses after 6 years. At implant insertion, both a 0.58-mm buccal bone on the platform and a 0.78-mm buccal bone at 2 mm apical to the implant shoulder are necessary for longer term maintenance of bone on the platform of implants specifically supporting CTI-FDPs.

Implant Treatment in Patient with Periodontitis: A Case Report with 13-year Follow-up.

This report describes long-term implant treatment in a patient with chronic periodontitis. The patient was a 59-year-old man who attended our facility requesting a dental implant. An initial examination revealed generalized gingival inflammation and subgingival calculus. Clinical examination revealed 55.3% of sites with a probing depth (PD) of >4 mm and 41.3% of sites with bleeding on probing. Radiographic examination revealed vertical bone resorption in #23, #33, #33, #35, and #47. Initial periodontal therapy consisting of plaque control, scaling and root planing, and tooth extraction was subsequently performed based on a clinical diagnosis of severe chronic periodontitis. Open flap debridement was performed for teeth with a PD >5 mm (#21, #22, #23, 333, #34, #35 and #47). After confirming the stability of the periodontal tissue, 3 implants were first placed in the maxilla (#25, #26, and #27). Final prostheses comprising a screw retaining-type implant superstructure were then placed (#25, #26, and 327). Following reevaluation, the patient was placed on supportive periodontal therapy. At 15 years after the first visit, the periodontal and implant conditions have remained stable. These results indicate that periodontal treatment before implantation and subsequent maintenance yield a clinically favorable and long-lasting outcome.

Therapeutic downregulation of neuronal PAS domain 2 (Npas2) promotes surgical skin wound healing.

Attempts to minimize scarring remain among the most difficult challenges facing surgeons, despite the use of optimal wound closure techniques. Previously, we reported improved healing of dermal excisional wounds in circadian clock neuronal PAS domain 2 (Npas2)-null mice. In this study, we performed high-throughput drug screening to identify a compound that downregulates Npas2 activity. The hit compound (Dwn1) suppressed circadian Npas2 expression, increased murine dermal fibroblast cell migration, and decreased collagen synthesis in vitro. Based on the in vitro results, Dwn1 was topically applied to iatrogenic full-thickness dorsal cutaneous wounds in a murine model. The Dwn1-treated dermal wounds healed faster with favorable mechanical strength and developed less granulation tissue than the controls. The expression of type I collagen, Tgfß1, and α-smooth muscle actin was significantly decreased in Dwn1-treated wounds, suggesting that hypertrophic scarring and myofibroblast differentiation are attenuated by Dwn1 treatment. NPAS2 may represent an important target for therapeutic approaches to optimal surgical wound management.

Rat peri-implant soft tissue specifically expressed CXCL2 on titanium implant during wound healing.

Many of genes specifically expressed in peri-implant soft tissue (PIST) selected by microarray analysis are involved in the inflammatory response. This study investigated the gene expression and localization of PIST-specific inflammatory markers in PIST during wound healing. Pure titanium implants were implanted into the rat upper mandibular socket to create PIST. Samples were harvested from PIST as an experimental group, and tooth extracted area of oral mucosa tissue (OMT) and healthy periodontal tissue (PT) as control groups. The gene expressions of four standard inflammatory markers and nine PIST-specific inflammatory markers including chemokine (C-X-C motif) ligand 2 (CXCL2) during wound healing were examined. Immunoreactions of CXCL2 and immune cells in PIST and control tissues were compared. During wound healing, gene expression of PIST-specific inflammatory markers was higher in PIST than in OMT (p < .05), but there were no significant differences in the expression of standard inflammatory markers. The molecule CXCL2 was expressed locally at the implant-connective tissue interface, and localization of immune cells closely matched the CXCL2 expression pattern. In PIST, seven of PIST-specific inflammatory markers were expressed specifically and strongly during wound healing and their expression was maintained until the end of healing. Furthermore, CXCL2 expression was due to the creation of the implant-connective tissue interface, and it established a unique defense mechanism in PIST that was not apparent in OMT or PT.

Clinical evaluation of periodontal pathogen levels by real-time polymerase chain reaction in peri-implantitis patients.

OBJECTIVE: The mechanisms underlying the onset and progression of peri-implantitis are similar to those of periodontitis, and the causative bacteria are believed to similar. Previous studies support an association between peri-implantitis and periodontal pathogen. Thus, we investigated the bacterial flora of peri-implantitis patients in comparison to those of healthy implant and periodontitis patients. MATERIALS AND METHODS: In total, 70 patients visiting Tokyo Dental College Chiba Hospital were divided into four groups: healthy, periodontitis, healthy implant, and peri-implantitis. For each group, the following five periodontal pathogens were detected using real-time polymerase chain reaction: Porphyromonas gingivalis, Aggregatibacter actinomycetemcomitans, Tannerella forsythia, Treponema denticola, and Prevotella intermedia. RESULTS: The average copy number of total bacteria was significantly higher in the periodontitis group than in the other groups. P. gingivalis was detected in the periodontitis and peri-implantitis groups at levels as high as 18.92% and 12.29%, respectively, and P. intermedia was found in the peri-implantitis group at a rate of 2.06%. Nevertheless, periodontal pathogens were generally detected at lower levels in the peri-implantitis group than in the periodontitis group. CONCLUSION: We found lower bacterial counts in the peri-implantitis group relative to the periodontitis group. Our results suggest that the peri-implant tissue is less resistant to bacteria, so even a small number of bacteria can be a risk factor for peri-implantitis and the causative agent of peri-implantitis can be bacteria other than periodontal pathogen.

Design and Synthesis of Cathepsin-K-Activated Osteoadsorptive Fluorogenic Sentinel (OFS) Probes for Detecting Early Osteoclastic Bone Resorption in a Multiple Myeloma Mouse Model.

We describe the design and synthesis of OFS-1, an Osteoadsorptive Fluorogenic Sentinel imaging probe that is adsorbed by hydroxyapatite (HAp) and bone mineral surfaces, where it generates an external fluorescent signal in response to osteoclast-secreted cathepsin K (Ctsk). The probe consists of a bone-anchoring bisphosphonate moiety connected to a Förster resonance energy transfer (FRET) internally quenched fluorescent (IQF) dye pair, linked by a Ctsk peptide substrate, GHPGGPQG. Key structural features contributing to the effectiveness of OFS-1 were defined by structure-activity relationship (SAR) and modeling studies comparing OFS-1 with two cognates, OFS-2 and OFS-3. In solution or when preadsorbed on HAp, OFS-1 exhibited strong fluorescence when exposed to Ctsk (2.5-20 nM). Time-lapse photomicrographs obtained after seeding human osteoclasts onto HAp-coated well plates containing preadsorbed OFS-1 revealed bright fluorescence at the periphery of resorbing cells. OFS-1 administered systemically detected early osteolysis colocalized with orthotopic engraftment of RPMI-8226-Luc human multiple myeloma cells at a metastatic skeletal site in a humanized mouse model. OFS-1 is thus a promising new imaging tool for detecting abnormal bone resorption.

Adhesion of streptococci to titanium and zirconia.

The purpose of this study was to evaluate the adherence of streptococci to disks of titanium (commercially pure titanium: CpTi) and zirconia (tetragonal zirconia polycrystals: TZP). CpTi and yttria-stabilized TZP disks with a mirror-polished surface were used as specimens. The arithmetic mean surface roughness (Ra and Sa) and the surface wettability of the experimental specimens were measured. For analyzing the outermost layer of the experimental specimens, X-ray photoelectron spectroscopy (XPS) analysis was performed. Streptococcus sanguinis, S. gordonii, S. oralis, and S. mutans were used as streptococcal bacterial strains. These bacterial cultures were grown for 24 h on CpTi and TZP. The number of bacterial adhesions was estimated using an ATP-bioluminescent assay, and scanning electron microscope (SEM) observation of the adhered bacterial specimens was performed. No significant differences in surface roughness or wettability were found between CpTi and TZP. In XPS analyses, outermost layer of CpTi included Ti0 and Ti4+, and outermost layer of TZP included Zr4+. In the cell adhesion assay, the adherences of S. sanguinis, S. gordonii, and S. oralis to TZP were significantly lower than those to CpTi (p < 0.05); however, significant difference was not observed for S. mutans among the specimens. The adherence to CpTi and TZP of S. mutans was significantly lower than that of S. sanguinis, S. gordonii, and S. oralis. These results were confirmed by SEM. S. sanguinis, S. gordonii, and S. oralis adhered less to TZP than to CpTi, but the adherence of S. mutans was similar to both surfaces. S. mutans was less adherent compare with the other streptococci tested in those specimens.

Neuronal PAS Domain 2 (Npas2)-Deficient Fibroblasts Accelerate Skin Wound Healing and Dermal Collagen Reconstruction.

The circadian clock, which consists of endogenous self-sustained and cell-autonomous oscillations in mammalian cells, is known to regulate a wide range of peripheral tissues. The unique upregulation of a clock gene, neuronal PAS domain protein 2 (Npas2), observed along with fibroblast aging prompted us to investigate the role of Npas2 in the homeostasis of dermal structure using in vivo and in vitro wound healing models. Time-course healing of a full-thickness skin punched wound exhibited significantly faster wound closure in Npas2-/- mice than wild-type (WT) C57Bl/6J mice. Dorsal skin fibroblasts isolated from WT, Npas2+/-, and Npas2-/- mice exhibited consistent profiles of core clock gene expression except for Npas2 and Per2. In vitro behavioral characterizations of dermal fibroblasts revealed that Npas2-/- mutation was associated with increased proliferation, migration, and cell contraction measured by floating collagen gel contraction and single-cell force contraction assays. Npas2 knockout fibroblasts carrying sustained the high expression level of type XII and XIV FAICT collagens and synthesized dermis-like thick collagen fibers in vitro. Confocal laser scanning microscopy demonstrated the reconstruction of dermis-like collagen architecture in the wound healing area of Npas2-/- mice. This study indicates that the induced Npas2 expression in fibroblasts may interfere with skin homeostasis, wound healing, and dermal tissue reconstruction, providing a basis for novel therapeutic target and strategy. Anat Rec, 2019. © 2019 Wiley Periodicals, Inc.

The characteristic regulation of gene expression Lbp and Sod3 in peri-implant connective tissue of rats.

The aim of this study was to investigate the characteristic gene expression profile and localization of peri-implant connective tissue (PICT) compared with those of periodontal connective tissue (PCT) and oral mucosal connective tissue (OMCT). Upper first molar of 5-week-old rats were extracted and titanium implant were placed for PICT group. PCT and OMCT were used as control. Laser microdissected connective tissue at 4 weeks used for microarray analysis. The expression and localization of selected genes were confirmed by quantitative real-time PCR (qRT-PCR) and immunohistochemistry. Approximately, 1000 genes of upregulated and downregulated in PICT compared with PCT and OMCT were recognized. Based on the results of microarray analysis and qRT-PCR were demonstrated lipopolysaccharide binding protein (Lbp) as a specific upregulated gene and superoxide dismutase 3 (Sod3) as a specific downregulated gene in PICT. Immunoreaction of LBP and F4/80 as macrophage marker localized to subepithelial and implant facing connective tissue in PICT. SOD3 expression was not observed in PICT, reactive oxygen species, a target of superoxide dismutase, was strongly and locally expressed in all three tissues. Our data suggested that the upregulation of Lbp and downregulation of Sod3 are as characteristic gene expression pattern in PICT.

Rescue bisphosphonate treatment of alveolar bone improves extraction socket healing and reduces osteonecrosis in zoledronate-treated mice.

Bisphosphonate (BP)-related osteonecrosis of the jaw, previously known as BRONJ, now referred to more broadly as medication-related osteonecrosis of the jaw (MRONJ), is a morbid condition that represents a significant risk for oncology patients who have received high dose intravenous (IV) infusion of a potent nitrogen containing BP (N-BP) drug. At present, no clinical procedure is available to prevent or effectively treat MRONJ. Although the pathophysiological basis is not yet fully understood, legacy adsorbed N-BP in jawbone has been proposed to be associated with BRONJ by one or more mechanisms. We hypothesized that removal of the pre-adsorbed N-BP drug common to these pathological mechanisms from alveolar bone could be an effective preventative/therapeutic strategy. This study demonstrates that fluorescently labeled BP pre-adsorbed on the surface of murine maxillo-cranial bone in vivo can be displaced by subsequent application of other BPs. We previously described rodent BRONJ models involving the combination of N-BP treatment such as zoledronate (ZOL) and dental initiating factors such as tooth extraction. We further refined our mouse model by using gel food during the first 7 days of the tooth extraction wound healing period, which decreased confounding food pellet impaction problems in the open boney socket. This refined mouse model does not manifest BRONJ-like severe jawbone exposure, but development of osteonecrosis around the extraction socket and chronic gingival inflammation are clearly exhibited. In this study, we examined the effect of benign BP displacement of legacy N-BP on tooth extraction wound healing in the in vivo model. Systemic IV administration of a low potency BP (lpBP: defined as inactive at 100 µM in a standard protein anti-prenylation assay) did not significantly attenuate jawbone osteonecrosis. We then developed an intra-oral formulation of lpBP, which when injected into the gingiva adjacent to the tooth prior to extraction, dramatically reduced the osteocyte necrosis area. Furthermore, the tooth extraction wound healing pattern was normalized, as evidenced by timely closure of oral soft tissue without epithelial hyperplasia, significantly reduced gingival inflammation and increased new bone filling in the extraction socket. Our results are consistent with the hypothesis that local application of a rescue BP prior to dental surgery can decrease the amount of a legacy N-BP drug in proximate jawbone surfaces below the threshold that promotes osteocyte necrosis. This observation should provide a conceptual basis for a novel strategy to improve socket healing in patients treated with BPs while preserving therapeutic benefit from anti-resorptive N-BP drug in vertebral and appendicular bones.

Neuronal PAS domain 2 (Npas2) facilitated osseointegration of titanium implant with rough surface through a neuroskeletal mechanism.

Titanium (Ti) biomaterials have been applied to a wide range of implantable medical devices. When placed in bone marrow, Ti-biomaterials integrate to the surrounding bone tissue by mechanisms that are not fully understood. We have previously identified an unexpected upregulation of circadian clock molecule neuronal PAS domain 2 (Npas2) in successfully integrated implant with a rough surface. This study aimed to elucidate the molecular mechanism of osseointegration through determining the role of Npas2. Human bone marrow stromal cells (BMSC) that were cultured on a Ti disc with SLA surface exhibited increased NPAS2 expression compared to BMSC cultured on a machined surface. A mouse model was developed in which miniature Ti implants were surgically placed into femur bone marrow. The implant push-out test and bone-to-implant contact measurements demonstrated the establishment of osseointegration in 3 weeks. By contrast, in Npas2 functional knockout (KO) mice, the implant push-out value measured for SLA surface Ti implant was significantly decreased. Npas2 KO mice demonstrated normal femur bone structure surrounding the Ti implant; however, the recovered implants revealed abnormal remnant mineralized tissue, which lacked dense collagen architecture typically found on recovered implants from wild type mice. To explore the mechanisms leading to the induced Npas2 expression, an unbiased chemical genetics analysis was conducted using mouse BMSC carrying an Npas2-reporter gene for high throughput screening of Library of Pharmacologically Active Compounds. Npas2 modulating compounds were found clustered in regulatory networks of the α2-adrenergic receptor and its downstream cAMP/CREB signaling pathway. Mouse primary BMSC exposed to SLA Ti disc significantly increased the expression of α2-adrenergic receptors, but the expression of ß2-adrenergic receptor was unaffected. Our data provides the first evidence that peripheral clock gene component Npas2 plays a role in facilitating the enhanced osseointegration through neuroskeletal regulatory pathways induced by BMSC in contact with rough surface Ti implant.

Bone marrow stromal cells from low-turnover osteoporotic mouse model are less sensitive to the osteogenic effects of fluvastatin.

This study aimed to investigate the effects of fluvastatin on the differentiation of bone marrow stromal cells (BMSCs) into osteoblasts in senescence-accelerated mouse prone 6 (SAMP6) compared with that in the normal senescence-accelerated-resistant mouse (SAMR1) model. SAMP strains arose spontaneously from the AKR/J background and display shortened life span and an array of signs of accelerated aging, compared with control SAMR strains. The dose effects of fluvastatin were also evaluated. BMSCs were cultured with/without fluvastatin (0 µM, 0.1 µM, 0.5 µM, and 1.0 µM). WST-1-based colorimetry was performed to evaluate cell proliferation. To evaluate cell differentiation, gene expression levels of bmp2 and runx2 were determined by quantitative reverse transcription polymerase chain reaction (qRT-PCR), and protein expression levels were determined using enzyme-linked immunosorbent assay (BMP2) and immunofluorescence staining (BMP2 and Runx2). Alkaline phosphatase (ALP) activity assay and histochemical detection were determined; the effect of noggin, a BMP-specific antagonist, was examined using ALP histochemical detection. To assess for mature osteogenic marker, gene expression levels of bglap2 were determined by qRT-PCR and mineralization was determined by alizarin red staining. RhoA activity was also examined by Western blotting. In SAMP6, BMP2, Runx2 and Bglap2 mRNA and protein expressions were significantly increased by fluvastatin, and ALP activity was increased by BMP2 action. RhoA activity was also inhibited by fluvastatin. The concentration of fluvastatin sufficient to increase BMP2 and Runx2 expression and ALP activity was 0.5 µM in SAMP6 and 0.1 µM in SAMR1. In conclusion, the present study revealed that fluvastatin promoted BMSC differentiation into osteoblasts by RhoA-BMP2 pathway in SAMP6. BMSCs of SAMP6 are less sensitive to the osteogenic effects of fluvastatin than SAMR1.

Dental Implant Treatment with Computer-assisted Surgery for Bilateral Agenesis of Maxillary Lateral Incisors: A Case Report.

Here, we report a case of dental implant treatment involving computer-assisted surgery for bilateral agenesis of the maxillary lateral incisors. The patient was a 39-year-old woman with the chief complaint of functional and esthetic disturbance due to maxillary and mandibular malocclusion. The treatment plan comprised non-extraction comprehensive orthodontic treatment and prosthodontic treatment for space due to the absence of bilateral maxillary lateral incisors. A preliminary examination revealed that the mesiodistal spaces left by the absent bilateral maxillary lateral incisors were too narrow for implant placement (right, 5.49 mm; left, 5.51 mm). Additional orthodontic treatment increased these spaces to approximately 6 mm, the minimum required for implant placement if risk of damage to the adjacent teeth due to inaccuracies in directionality of drilling is to be avoided. For dental implant treatment with computer-assisted surgery, preoperative planning/simulation was performed using Simplant® ver.12 software and a toothsupported surgical template fabricated using stereolithography. Two narrow-diameter implants were placed in a two-stage procedure. It was confirmed that there was sufficient distance between the implant fixtures and the roots of the adjacent teeth, together with no exposure of alveolar bone. Following a 4-month non-loading period, second-stage surgery and provisional restoration with a temporary screw-retained implant crown were performed. Cement-retained superstructures made of customized zirconia abutment and a zirconia-bonded ceramic crown were fitted as the final restoration. At 5 years after implant surgery, there were no complications, including inflammation of the peri-implant soft tissue and resorption of peri-implant bone. Computer-assisted implant surgery is useful in avoiding complications in bilateral agenesis of the maxillary lateral incisors when only a narrow mesiodistal space is available for implant placement.

Release properties of atelocollagen-gelatin complexes as carriers for local administration of fluvastatin.

The aim of this study was to investigate properties of atelocollagen/gelatin complexes (AC/Gel) and their characteristics of sustained statin release, to assess the utility of AC/Gel. AC/Gel were prepared by changing the mixing ratio of AC (0 to 40% of AC). Analysis of spectra of fluvastatin (Flu), gelatin (Gel), and Flu with Gel complex using a Fourier transform-infrared spectrometer indicates that Flu was bound to Gel through a bond involving the carboxyl and amino groups. Evaluation of characteristics of sustained release of Flu from the AC/Gel using an ultraviolet-visible spectrophotometer showed that the release rate of Flu decreased with increasing the AC content. The histological evaluation using of Sprague-Dawley rats suggest that, unlike the pure Gel sponge, the AC/Gel was not absorbed in an early stage. Therefore, the present study showed that sustained Flu release can be controlled by using an AC/Gel, suggesting the utility of this composite material.

The genes Scgb1a1, Lpo and Gbp2 characteristically expressed in peri-implant epithelium of rats.

OBJECTIVES: The peri-implant epithelium (PIE) plays an important role in the prevention against initial stage of inflammation. To minimize the risk of peri-implantitis, it is necessary to understand the biological characteristics of the PIE. The aim of this study was to investigate the characteristic gene expression profile of PIE as compared to junctional epithelium (JE) using laser microdissection and microarray analysis. METHODS: Left upper first molars of 4-week-old rat were extracted, and titanium alloy implants were placed. Four weeks after surgery, samples were harvested by laser microdissection, and total RNA samples were isolated. Comprehensive analyses of genes expressed in the JE and PIE were performed using microarray analysis. Confirmation of the differential expression of selected genes was performed by quantitative real-time polymerase chain reaction and immunohistochemistry. RESULTS: The microarray analysis showed that 712 genes were more than twofold change upregulated in the PIE compared with the JE. Genes Scgb1a1 were significantly upregulated more than 19.1-fold, Lpo more than 19.0-fold, and Gbp2 more than 8.9-fold, in the PIE (P < 0.01). Immunohistochemical localization of SCGB1A1, LPO, and GBP2 was observed in PIE. CONCLUSION: The present results suggested that genes Scgb1a1, Lpo, and Gbp2 are characteristically expressed in the PIE.