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Various techniques have been applied to the fabrication of duplicate dentures. In this new method, after scanning the cameo and intaglio surfaces of the complete denture using an intraoral scanner, the two images were superimposed on the interocclusal image. The linear gap between cameo and intaglio surfaces was filled using free downloadable CAD software. The superimposed image was subsequently used as data for fabricating the duplicate complete denture with a 3D printer. Using the intraoral scanner, duplication of complete dentures could be performed without needing any complicated processes, laboratory scanners, or advanced CAD software.

Purpose To investigate the difference in improvement of oral health-related quality of life (OHR-QoL) depending on the oral and denture conditions of a complete denture wearer when using a cream or powder type denture adhesive in a 10-center parallel randomized clinical trial.Methods Two hundred edentulous subjects who wore complete dentures were allocated to each of the three groups according to denture adhesive type: cream, powder, and control (saline solution). The materials were applied to the mucosal surface of the dentures for 4 days, and baseline data and data after the intervention were collected. OHR-QoL was assessed using the Japanese version of the modified Oral Health Impact Profile for Edentulous Patients (OHIP-EDENT-J) scale for outcome. Multivariate analysis was used to investigate improvements in OHR-QoL according to participant characteristics among complete denture wearers using these materials.Results OHIP-EDENT-J scores were significantly decreased in all groups after the intervention (p < 0.05); however, there were no statistically significant differences among the groups. Multiple logistic regression analysis revealed a significant association between the vertical height of the maxillary and mandibular alveolar ridge and OHIP-EDENT-J scores in the cream-type denture adhesive group. In contrast, there were no significant association between participant characteristics and OHIP-EDENT-J scores in the powder-type adhesive and control groups.Conclusions The use of denture adhesives could improve OHR-QoL for complete denture wearers. The cream-type denture adhesives may be expected to improve OHR-QoL in patients with poor residual ridge conditions compared with patients with good residual ridge conditions.

Abstract A DNA hybridization-based differential peptide display (DPD) was developed for the screening of phage peptide library to find osteogenic peptides intended to bind to epigenetically induced osteogenic receptors on NIH/3T3 (3T3) cell surface. In the presence of DNA methylation inhibitor of 5-azacytidine (5AZC), an osteoblastic receptor of bone morphogenetic protein (BMP) receptor 1A (BMPR1A) was induced on the cell surface of NIH/3T3 fibroblasts. Cyclic heptamer-displaying phage library was screened against vehicle and 5AZC treated (Tx) 3T3 cells. Antisense oligo against library against library peptide coding DNA of control 3T3 cell bound phages were synthesized to subtract common binders from that of 5AZC-Tx 3T3 cell-bound phages that included 5AZC-induced receptor binders. The library peptide coding regions of conformational receptor binder-subtracted DPD were PCR-amplified and cloned into a plasmid vector specifically designed for short peptide expression. No unique binder was identified when 96 clones were randomly picked from the third round of panning against 5AZC-treated 3T3 cells, suggesting miscellaneous bindings to cell surface proteins. Unique binders showing homology to known function proteins were successfully identified when constitutive receptor binders were subtracted from 5AZC-induced protein binders. Some of identified peptides significantly increased alkaline phosphatase activity in 5AZC-Tx 3T3 cells. DPD can be a useful tool to screen functional peptide bindings to cell surface receptors. Graphic Abstract

The purpose of this study was to investigate the effect of denture adhesives on oral moisture in a 10-center parallel randomized clinical trial.Two hundred edentulous subjects wearing complete dentures were allocated into three groups: cream-type adhesive, powder-type adhesive and control groups. The adhesives (and saline solution in the control group) were applied to the mucosal surface of the dentures for 4 days, and baseline data and data after the intervention for eight meals over 4 days were obtained. For the main outcome, oral moisture was measured with a moisture checking device. Secondary outcomes were denture satisfaction, masticatory performance, denture retention, and occlusal force. In addition to between-group and within-group comparisons of oral moisture, investigations for secondary outcomes were undertaken in subgroups classified according to the degree of oral moisture at baseline (normal subgroup and dry mouth subgroup). Intention-to-treat analysis was also performed.Between-group and within-group comparisons of oral moisture showed no significant differences. The cream-type and powder-type denture adhesives were significantly effective in the dry mouth group for denture satisfaction ratings of ability to masticate, stability, retention, and comfort of mandibular dentures (p0.05). The masticatory performance and retentive force of the dry mouth denture adhesive using groups were significantly improved after intervention (p0.05).The oral moisture of complete denture wearers was not influenced by the use of denture adhesives. Our findings showed that denture adhesives improved subjective denture satisfaction, masticatory performance, and retention for complete denture patients with oral dryness.

Titanium is often used in the medical field and in dental implants due to its biocompatibility, but it has a high rate of leading to peri-implantitis, which progresses faster than periodontitis. Therefore, in the present study, the expression of cytokines from gingival epithelial cells by nanotitania was investigated, which is derived from titanium in the oral cavity, and the additional effect of Porphyromonasgingivalis (periodontopathic bacteria) lipopolysaccharide (PgLPS) was investigated. Ca9-22 cells were used as a gingival epithelial cell model and were cultured with nanotitania alone or with PgLPS. Cytokine expression was examined by reverse transcription-quantitative polymerase chain reaction and enzyme-linked immunosorbent assay. In addition, cellular uptake of nanotitania was observed in scanning electron microscopy images. The expression of interleukin (IL)-6 and IL-8 significantly increased in Ca9-22 cells by nanotitania treatment alone, and the expression was further increased by the presence of PgLPS. Nanotitania was observed to phagocytose Ca9-22 cells in a dose- and time-dependent manner. Furthermore, when the expression of IL-11, related to bone resorption, was investigated, a significant increase was confirmed by stimulation with nanotitania alone. Therefore, nanotitania could be associated with the onset and exacerbation of peri-implantitis, and the presence of periodontal pathogens may worsen the condition. Further clinical reports are needed to confirm these preliminary results.

When the sinus is enlarged, it may be necessary to elevate the floor of the maxillary sinus using the crestal or lateral approach. This report presents a case where sinus floor elevation was performed using tricalcium phosphate (TCP) plates supported by implant bodies, and only the blood clot was present around the implant bodies. Cone-beam computed tomography images, taken one year after the lateral approach, revealed the presence of a TCP-like radio-opacity, which almost disappeared after two years. About seven years after the surgery, the patient's superstructure and occlusion were stable. Furthermore, the grafted TCP was completely absorbed, and the implant body appeared to be in good condition, with no signs of bone resorption around the implant.

Purpose To reveal effects of improvement of masseter muscle activity on brain function in elderly people wearing complete dentures.Methods Subjects were 14 edentulous patients with a chief complaint of the inconvenience of their complete dentures. The surface electromyographic (EMG) activity of the masseter muscles was measured. Brain activities were analyzed with functional magnetic resonance imaging (fMRI), employing chewing gum as the task program. Cognitive functions were evaluated with Trail Making Test Part A (TMT-A), Rey Auditory Verbal Learning Test (RAVLT) and Rey-Osterrieth Complex Figure Test (R-OCFT). Those evaluations were performed in which subjects wore their old dentures (OD) or newly fabricated dentures (ND).Results We compared ND condition with OD condition. The masseter muscle activity significantly increased in ND condition (p0.05, Wilcoxon signed rank test). The brain activity increased significantly in the superior frontal gyrus, precentral gyrus, putamen, inferior parietal lobule, cerebellum, inferior frontal lobe, and middle frontal gyrus under the ND condition than under the OD condition (p0.01, uncorrected, cluster size10 voxels). Results of TMT-A, RAVLT, and R-OCFT were also significantly improved (p0.05, Wilcoxon signed rank test).Conclusions In the edentulous elderly, the brain activity was increased following the improvement of the masseter muscle activity. Consequently, it is possible that the improvement of the masseter muscle activity might influence on the attention, verbal skills, and visual memory.

PURPOSE The purpose of this study was to examine morphological changes in the superstructure of implants in relation to the degree of attrition of the implant superstructure and its antagonists. METHODS Thirty-three patients participated. The implant superstructures of the first molar and its antagonists were scanned with an intraoral scanner every 3 months. The amount of attrition was calculated in relation to the various materials used for the superstructure, and differences were analyzed statistically (P < 0.05). RESULTS Attrition of the implant superstructure was 110 ± 30 µm for monolithic zirconia and 105 ± 27 µm for resin-veneered metal after 12 months. No statistically significant difference was found between the two groups. In contrast, attrition of the antagonists was 75 ± 25 µm for zirconia and 105 ± 20 µm for resin-veneered metal after 12 months. A statistically significant difference in the attrition levels between the two groups was observed after 9 and 12 months. CONCLUSION During the 12-month observation period, there was no difference in superstructure attrition between zirconia and resin-veneered metal. Attrition of the antagonists was higher for hybrid composite resin-veneered metal than for zirconia, suggesting that surface roughness affected the degree of attrition.

Adequate oral status and functional assessments are important for dysphagia rehabilitation in acute care inpatient settings, especially to establish individualised oral intake. However, the association between food intake levels and oral function has not been elucidated in acute care inpatients. This cross-sectional study clarified the association between oral intake levels and the oral status/function of patients with dysphagia admitted to acute care settings. Admitted patients aged ≥40 years (n = 459; men: 288; mean age: 70.8 ± 12.0) examined at the Department of Dysphagia Rehabilitation at the Iwate Medical University Hospital from April 2007 to March 2014 were included. The oral health status was evaluated by the tongue coating, oral dryness severity, plaque control, posterior occlusal support and a repetitive saliva swallowing test (RSST). Dysphagia severity was determined from the Dysphagia Severity Scale. Oral intake levels were evaluated using the Functional Oral Intake Scale (FOIS) at the time of the initial dental examination (FOIS-I), and they were re-evaluated after the revision of levels according to the participants' general condition and oral health status (FOIS-R). Divergence between FOIS-I and FOIS-R was noted in >40% patients. Multiple regression analysis showed significant associations between FOIS-R and consciousness level, activities of daily living, tongue coating, RSST and posterior occlusal support. Patients with dysphagia in acute care settings require detailed assessments of their oral status and function, including swallowing, to determine the most suitable feeding methods and dental interventions to improve oral intake levels.

Purpose One of the most prominent issues in a super-aging society is the rapid increase in dementia patients. Cross-sectional studies in dentistry have indicated that patients with dementia have worse oral health compared to healthy people. The purpose of this study was to clarify the influence of tooth loss on brain structure by comparing the volumes of gray matter (GM) and white matter (WM) between edentulous and dentulous subjects. Methods Subjects were recruited from the Denture Clinic at Iwate Medical University Hospital Dental Center. Experiments were performed on edentulous (5 males, 8 females, 81.8±1.24years) and dentulous subjects (4 males, 7 females, 77.1±4.25years). Patients with dementia were excluded from this study. Brain volumes of GM and WM in edentulous and dentulous subjects were compared using intracranial volume, age, gender and history of hypertension as covariates. Analyzed brain areas were identified by transforming the Montreal Neurological Institute coordinate into the anatomical coordinate in edentulous subjects. Results The analysis of WM structural images found no morphological differences between dentulous and edentulous subjects. However, significant atrophy of GM was observed in the hippocampus, caudate nucleus and temporal pole of the right hemisphere in edentulous subjects. Conclusions The results of this study suggest that tooth loss was a causal factor for volume reduction in brain areas related to memory, learning and cognition.

Purpose The digitization of the dental field has been vigorously promoted in recent years. An impression using an intraoral scanner is considered to significantly change future dental treatment. The purpose of this review is to evaluate accuracy and practicality of various intraoral scanners and verification method of intraoral scanners. Study selection This review was based on articles searched through the MEDLINE and PubMed databases. The main keywords that were employed during the search were “Oral Scanner, Intraoral Scanners, Desktop Scanner, and Digital Impression”. Result It was reported that illuminance and color temperature affected trueness and precision of intraoral scanners. The repeatability of intraoral scanners indicated the possibility of producing fixed prostheses within the range of being partially edentulous. It is considered difficult to use intraoral scanners in fabricating cross-arch fixed prostheses. However, with intraoral scanners, it may be considered possible to fabricate mouth guards and dentures equivalent to those of desktop scanners. Current intraoral scanner scans are considered more comfortable than traditional impressions that use irreversible hydrocolloid and elastomeric impression materials. Conclusion Since the intraoral scanner is an evolving device, further improvement in accuracy is expected in the future. In addition, verification of the accuracy of intraoral scanners must be conducted accordingly.

Natural teeth are supported by connective tissue collagen fibers that insert perpendicularly in the tooth cementum. Perpendicular insertion plays an important role in the maintenance of the junction between the oral epithelium and the periodontal connective tissue. Most titanium dental implant surfaces have no micro or macro structure to support perpendicularly oriented collagen attachment. Without this tight biologic seal to resist bacterial invasion and epithelial downgrowth, progressive bone loss in peri-implantitis is seen around dental implants. The purpose of this study was to establish the perpendicularly oriented collagen attachment to titanium oxide nanotube (TNT), and to assess its binding stability. TNT was prepared on the titanium-surface by anodization. Scanning electron microscopy (SEM) showed a regularly aligned TNT with an average 67 nm-diameter when anodized at 30 V for 3 h. Subsequently, collagen type I (CoI) was electrophoretically fused to anodic TNT in native polyacrylamide gel system where negatively charged CoI-C term was perpendicularly navigated to TNT. SEM and atomic force microscopy (AFM) were used to analyze CoI on the TiO2 and TNT surface. Several tens of nanometers of CoI protrusion were recorded by AFM. These protrusions may be long enough to be priming sites for cell-secreted CoI. CoI laid parallel to the titanium surface when fused by a chemical linker. Binding resistance of CoI against drastic ultrasonication was measured by Fourier-transform infrared spectroscopy attenuated total reflection (FTIR-ATR). The electrophoretically fused CoI in the titanium nanotube (TNT–CoIEPF) showed the significantly greatest binding resistance than the other groups (P

Purpose To determine the appropriate attachment and design of a denture base for mandibular implant overdenture (IOD), the oral mucosa pressure caused by mandibular implant overdentures was measured using edentulous jaw models with various attachments. Methods An experimental edentulous mandibular model with a 1.5-mm thick artificial oral mucosa was used. Two implants were placed at the area equivalent to the bilateral canines of an experimental jaw model. Locator attachments (LA), ball attachments (BA), magnetic attachments (MA), and round-bar attachments (R-BA) were fabricated. Six miniature pressure sensors were placed at the bilateral buccal premolar regions, bilateral buccal shelves, and bilateral lingual molar regions. A precision universal testing machine was used to apply dynamic repetitive loads of 50 N. The load points were the center of the model, which should represent bilateral mastication on both sides (bilateral load), and were equivalent to the left first molar, which should represent unilateral mastication (unilateral load). Statistical analysis was performed using one-way analysis of variance. Multiple comparisons were then performed using the Bonferroni post hoc test. P Results Under the bilateral load condition, the lower oral mucosa pressure value with BA was measured, compared to other attachments at all measurement sites. Under the unilateral load condition, the oral mucosa pressure value of BA was smaller than the other attachments at the measurement site on the loading side. Conclusions BA has exerted the greatest effects on support and bracing, suggesting that, BA is suitable for reducing oral mucosa pressure during mastication.

The direction of mesenchymal stem cell (MSC) differentiation is regulated by stimulation with various growth factors and cytokines. We recently established MSC lines, [transforming growth factor-β (TGF-β)-responsive SG‑2 cells, bone morphogenetic protein (BMP)-responsive SG‑3 cells, and TGF-β/BMP-non-responsive SG‑5 cells], derived from the bone marrow of green fluorescent protein-transgenic mice. In this study, to compare gene expression profiles in these MSC lines, we used DNA microarray analysis to characterize the specific gene expression profiles observed in the TGF-β-responsive SG‑2 cells. Among the genes that were highly expressed in the SG‑2 cells, we focused on vascular endothelial growth factor (VEGF) receptor 3 (VEGFR3), the gene product of FMS-like tyrosine kinase 4 (Flt4). We found that VEGF-C, a specific ligand of VEGFR3, significantly induced the cell proliferative activity, migratory ability (as shown by Transwell migration assay), as well as the phosphorylation of extracellular signal-regulated kinase (ERK)1/2 in the SG‑2 cells. Additionally, VEGF-C significantly increased the expression of prospero homeobox 1 (Prox1) and lymphatic vessel endothelial hyaluronan receptor 1 (Lyve1), which are lymphatic endothelial cell markers, and decreased the expression of osteogenic differentiation marker genes in these cells. By contrast, TGF-β significantly increased the expression of early-phase osteogenic differentiation marker genes in the SG‑2 cells and markedly decreased the expression of lymphatic endothelial cell markers. The findings of our study strongly suggest the following: i) that VEGF-C promotes the proliferative activity and migratory ability of MSCs; and ii) VEGF-C and TGF-β reciprocally regulate MSC commitment to differentiation into lymphatic endothelial or osteoblastic phenotypes, respectively. Our findings provide new insight into the molecular mechanisms underlying the regenerative ability of MSCs.

Although the effects of neprilysin (NEP), also called CD10, on the clearance of Alzheimer's disease (AD)-associated amyloid-β (Aβ) have been reported, NEP is not made in the brain, and the mechanism for the transport of NEP to the brain has not been investigated. Our hypothesis is that muscle packages NEP in exosomes in response to a neuromuscular signal and sends it to the brain via retrograde axonal transport. The masseter muscle (MM) and the trigeminal nerve (TGN) are good candidates for this mechanism by virtue of their proximity to the brain. The aim of this study was to trace the NEP protein from the MM, through the TGN, and to the hippocampus (HPC) in muscle contraction models in vitro and in vivo. NEP expression in mouse tissue lysates was analyzed by RT-PCR and Western blot. Four-week-old mice were perfused to remove blood NEP contamination. The MM expressed substantial levels of NEP protein and mRNA. On the other hand, a remarkably high level of NEP protein was measured in the TGN in the absence of mRNA. NEP protein, without the corresponding mRNA, was also detected in the HPC. These results suggested that the MM derived NEP was taken up by the TGN, which in turn permitted NEP access to the central nervous system and within it the HPC. When the MM was induced to contract by electric stimulation in freshly euthanized mice, NEP protein decreased in the MM in a stimulus time-dependent manner, while that in the TGN and the HPC increased sequentially. Furthermore, NIR-labeled exosomes tracked along the same route. Finally, carbachol induced secretion of exosomal NEP in C2C12-derived myotube cells. These results support our hypothesis that MM-derived NEP is transported along the TGN to reach the HPC following electrical or cholinergic stimulation.

Epithelial attachment via the basal lamina on the tooth surface provides an important structural defence mechanism against bacterial invasion in combating periodontal disease. However, when considering dental implants, strong epithelial attachment does not exist throughout the titanium-soft tissue interface, making soft tissues more susceptible to peri-implant disease. This study introduced a novel synthetic peptide (A10) to enhance epithelial attachment. A10 was identified from a bacterial peptide display library and synthesized. A10 and protease-activated receptor 4-activating peptide (PAR4-AP, positive control) were immobilized on commercially pure titanium. The peptide-treated titanium showed high epithelial cell migration ability during incubation in platelet-rich plasma. We confirmed the development of dense and expanded BL (stained by Ln5) with pericellular junctions (stained by ZO1) on the peptide-treated titanium surface. In an adhesion assay of epithelial cells on A10-treated titanium, PAR4-AP-treated titanium, bovine root and non-treated titanium, A10-treated titanium and PAR4-AP-treated titanium showed significantly stronger adhesion than non-treated titanium. PAR4-AP-treated titanium showed significantly higher inflammatory cytokine release than non-treated titanium. There was no significant difference in inflammatory cytokine release between A10-treated and non-treated titanium. These results indicated that A10 could induce the adhesion and migration of epithelial cells with low inflammatory cytokine release. This novel peptide has a potentially useful application that could improve clinical outcomes with titanium implants and abutments by reducing or preventing peri-implant disease.

Objective To investigate the effects of wearing complete dentures on pharyngeal shape for swallowing in edentulous older people. Background In the absence of complete dentures, edentulous older people often lose the occlusal support necessary to position the mandible, which leads to an anterosuperior shift of the mandible during swallowing. This may result in pharyngeal shape changes effecting swallowing function in older people. However, the details of this phenomenon are currently unclear. Materials and methods Participants were 17 older edentulous volunteers. Cone-beam computed tomography imaging was performed with the participant in the seated position and wearing (i) both maxillary and mandibular dentures, (ii) maxillary dentures only and (iii) no dentures. During imaging, participants were instructed to keep their mouth closed to the mandibular position determined in advance during swallowing for each denture-wearing condition. The volume, height and average cross-sectional area of the velopharynx and oropharynx were measured, and the positions of the epiglottis and mandible were recorded. Results While the vertical height of the oral cavity and pharynx significantly decreased, the volume and average cross-sectional area of the oropharynx significantly increased when dentures were not worn (p

The key objective of mastication is to form a food bolus suitable for smooth swallowing. However, chewing is usually performed without a conscious effort. Poor bolus formation can cause pharyngeal residue and suffocation in elderly individuals with reduced swallowing function. Therefore, chewing with a conscious effort may help the bolus to more easily pass the pharynx. This study aimed to clarify the impact of mastication with a conscious effort on the feeding sequence. Subjects included 25 dentulous volunteers who were informed and provided written consent. Lateral videofluoroscopy was performed during the feeding of solid agar jelly under two conditions: chewing naturally in their usual manner (without volition) and chewing with a conscious effort (with volition). Temporal evaluation was performed for mastication, stage II transport (STII), swallow onset and oropharyngeal transit time. Moreover, bolus volume at swallow onset and subjective evaluation of swallowing easiness were measured. Volitional chewing with a conscious effort lengthened the duration of the chewing sequence before and after STII and delayed the swallow onset despite the fact that the bolus volume in the vallecula and hypopharynx (HYP) had significantly increased. Furthermore, with volition, the bolus transit time from swallow onset in the oral cavity, upper oropharynx and HYP was reduced, and subjective evaluation of swallowing easiness demonstrated significant improvement. These results suggest that volitional chewing with a conscious effort can alter bolus transport and swallowing, resulting in easier swallowing.

This position paper proposes a definition and naming standard for removable partial dentures (RPDs) using thermoplastic resin, and presents a guideline for clinical application. A panel of 14 experts having broad experience with clinical application of RPDs using thermoplastic resin was selected from members of the Japan Prosthodontic Society. At a meeting of the panel, “non-metal clasp denture” was referred as the generic name of RPDs with retentive elements (resin clasps) made of thermoplastic resin. The panel classified non-metal clasp dentures into two types: one with a flexible structure that lacks a metal framework and the other having a rigid structure that includes a metal framework. According to current prosthetic principles, flexible non-metal clasp dentures are not recommended as definitive dentures, except for limited cases such as patients with a metal allergy. Rigid non-metal clasp dentures are recommended in cases where patients will not accept metal clasps for esthetic reasons. Non-metal clasp dentures should follow the same design principles as conventional RPDs using metal clasps.

Palatal coverage is often required for elderly edentulous patients with complete dentures. The purpose of this study was to clarify impacts of palatal coverage on bolus formation and subsequent adaptive changes. Subjects were 18 healthy young dentulous adults who wore 1·5-mm-thick palatal plates. Subjects were asked to feed 12 g of bicoloured rice as usual, and the bolus formation by mastication and swallowing in the pharynx was observed using a nasal videoendoscopy. The bolus formation index (BFI), number of mastication strokes until swallowing, visual analogue scale about swallowing easiness and masticatory performance using colour-changeable gum were measured under three conditions: before placement of the palatal plate (day 0), immediately after placement (day 1) and after 7 days of wearing the plate (day 7). BFI and visual analogue scale on day 1 were significantly lower than those on day 0, but those on day 7 significantly recovered to the level of day 0. The number of mastication strokes did not change from day 0 to day 1, however, that on day 7 was significantly higher. Masticatory performance on days 1 and 7 was significantly lower than that on day 0. Although palatal coverage inhibits bolus formation during feeding, subjects increased the number of mastication strokes until swallowing threshold as they adapted to palatal coverage over time. This adaptive change was due to compensate for the lowered masticatory performance to achieve bolus formation for comfortable swallowing.

Recently, we identified the scrapie responsive gene 1 (SCRG1) secreted from mesenchymal stem cells (MSCs) and its receptor bone marrow stromal cell antigen 1 (BST1) as positive regulators of stem cell qualities such as self-renewal, migration abilities, and osteogenic differentiation potential. Here, we examined the effect of the paracrine activity of SCRG1 in macrophages. The mouse macrophage-like cell line Raw264.7 expressed BST1/β1 or BST1/β2 integrin as possible SCRG1 receptors. Unexpectedly, recombinant SCRG1 did not enhance cell proliferation, migration, or adhesion in these macrophages. However, further examination of the effect of SCRG1 in Raw264.7 cells did reveal a potent anti-inflammatory effect whereby SCRG1 suppressed LPS-induced CCL22 production. SCRG1 also induced the phosphorylation of extracellular signal-regulated kinase 1/2 (ERK1/2) in these cells and, moreover, a mitogen-activated protein kinase (MAPK)/ERK kinase inhibitor U0126 significantly suppressed the effect of SCRG1 on LPS-induced chemokine CCL22 production. Taken together, these data indicate that SCRG1 signals through the MAPK pathway and suppresses the LPS signaling pathway. CCL22 is generally known to be chemotactic for monocytes, dendritic cells, natural killer cells and chronically activated T lymphocytes, suggesting that MSC-derived SCRG1 may block infiltration of these cells. A mechanism is proposed by which MSCs play their immunosuppressive role through suppressing chemokine expression in monocyte/macrophage lineage cells.

Wearing dentures and dysphagia are common in older individuals; however, it is still unknown how dentures affect oral and pharyngeal swallowing. The purpose of this study was to reveal the effects of wearing and removing dentures on oropharyngeal movements during pharyngeal swallowing in the feeding sequence of solid food. Participants were 25 edentulous volunteers (nine men, 16 women; mean age 76·2 years) who wore complete dentures. The test food was minced agar jelly containing barium sulphate. Subjects were instructed to feed and swallow the test food with or without dentures during observation using videofluorography. We quantitatively evaluated the range, distance and duration of oropharyngeal movements during pharyngeal swallowing. When dentures were absent, the range of mandible and hyoid movements were significantly expanded in the anterosuperior direction, and the range of laryngeal movement was significantly expanded in the anterior direction. Additionally, the posterior pharyngeal wall contraction and upper oesophageal sphincter opening significantly increased. In addition, the distances of the mandible, hyoid and laryngeal movements and the mandibular duration were significantly extended when dentures were absent. No significant differences were observed in the duration of movements of other organs between wearing and removing dentures. The hyoid bone, larynx, posterior pharyngeal wall and upper oesophageal sphincter do not change their duration of movements when dentures were removed but, rather, expand their range of movement. This might be a spatial change of oropharyngeal movement to avoid temporal changes in pharyngeal swallowing when dentures were absent in edentulous older individuals.

Background It is important to visualize the phenomenon which occurs in actual clinical cases to decide the timing of implant placement subsequently after sinus graft. Although several clinical cases of bone augmentation using xenograft have been evaluated, the number of those reports which have described bone remodeling by alloplasty are not sufficient. Findings In the present report, to investigate bone remodeling histologically after maxillary sinus augmentation with porous hydroxyapatite alloplast, bone cores from the sinus floor of three female nonsmoking patients (aged 64–73 years) were collected 6 months after sinus lift surgery, embedded in methyl methacrylate resin, and prepared by conventional methods. Bone architecture and graft residues were evaluated by micro-computed tomography of the same specimens. Hematoxylin–eosin and Villanueva–Goldner staining revealed mature osteoblasts and multinucleated osteoclasts on the grafted sinus floor and surface of residual hydroxyapatite particles. The particulate interspace was partially filled with osteoid and calcified tissue and showed active vascularization. Conclusion The results suggested that bone regeneration and angiogenesis within and between porous hydroxyapatite particles were sufficiently found after 6 month histologically in the grafted sinus floor. Electronic supplementary material The online version of this article (doi:10.1186/s40064-016-1885-2) contains supplementary material, which is available to authorized users.

Date Y, Yokoyama Y, Kondo H, Kuroda S, Ohya K, Ota MS, Iseki S, Kasugai S. Restricted expression of chromatin remodeling associated factor Chd3 during tooth root development. J Periodont Res 2012; 47: 180–187. © 2011 John Wiley & Sons A/S Background and Objective: The tooth root is one of the critical parts to maintain tooth function; however, the molecular mechanisms of root development remain unknown. We aimed to identify specific factors for root morphogenesis using a newly developed experimental system. Material and Methods: Tentative cementoblasts and periodontal ligament cells from mouse mandibular molars were isolated using laser capture microdissection. More than 500 cementoblasts and periodontal ligament cells were separately captured. After RNA extraction and amplification, mRNA expression in isolated cementoblasts was compared with that of periodontal ligament cells by cDNA microarray analysis. Then, putative cementoblast-specific genes were subjected to in situ hybridization analysis to confirm the results in mouse mandible. Results: Approximately 2000 genes were differentially expressed between these tissues. Among those genes, zinc finger helicase (ZFH), also termed chromodomain-helicase-DNA-binding protein 3 (Chd3), was one of the highly expressed transcripts in tentative cementoblasts. In situ hybridization revealed that ZFH/Chd3 was strongly expressed in Hertwig’s epithelial root sheath rather than in cementum. Moreover, its expression disappeared when root formation was advanced in the first molar. In contrast, Chd3 was continuously expressed in dental epithelial cells of the cervical loop, in which root extension is never terminated. Conclusion: These results suggest that ZFH/Chd3 might play an important role in tooth root development and subsequent cementogenesis.

Collagen containing bone morphogenetic protein-2 (BMP-2) expression vector, which is called “gene-activated matrix,” promotes bone regeneration when transplanted to the bone defect. We speculated that hydroxyapatite fiber (HF) would be an ideal matrix for “gene-activated matrix” especially for bone regeneration, because it is oseteoconductive and has high affinity to DNA. The purpose of this study is to clarify whether HF containing BMP-2 expression vector induces ectopic bone formation. We prepared HF containing 0, 10, 50, and 100 μg BMP-2 expression vector. Wistar male rats (8 weeks) were used and each rat received two HF implants in the left and right dorsal muscle. The rats were sacrificed 4, 8, and 12 weeks after the operation, and implants were analyzed radiographically by softex, dual-energy X-ray absorptiometry, and they were histologically examined. At 4 weeks, HF containing 50 or 100 μg BMP-2 expression vector showed high bone mineral contents and large radiopaque volume compared to the other implants. At 8 and 12 weeks, HF containing 50 μg BMP-2 expression vector exerted the highest values in the radiographic analyses. Bonelike tissue was histologically observed in HF containing 50 and 100 μg BMP-2 expression vector groups but not detected in the other implants. The present results suggest that HF is potential as a matrix for “gene-activated matrix” for bone tissue engineering. © 2008 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 2009

The detailed features of cementum and PDL remain unknown and specific markers for cemento- blasts and PDL cells have not been identified. Moreover, the molecular mechanism of periodontal tissue development, homeostasis and regeneration is also obscure. Since cementum or PDL tissue in previous studies was usually obtained by manual curettage, it has been difficult to isolate pure cementum or PDL. In the current study, laser capture microdissection(LCM)of undecalcified frozen sections of murine mandible was employed for precise tissue isolation and to obtain intact RNA from cementoblasts and PDL cells of the murine mandible. Over 500 tentative cementoblasts and PDL cells were individually captured using an infra- red laser. A bioanalyzer detected peaks of 18S and 28S rRNA, suggesting that the quality of RNA was sufficient. Subsequently, mRNA expression in cementoblasts was compared with that in PDL cells by GeneChip analysis, showing that approximately 2,000 genes were differently expressed between those tissues. Serial cementoblast-positive and PDL-negative genes were subjected to real-time RT-PCR analy- sis to support GeneChip results, using mandible and femur RNA samples. Several genes were expressed at higher levels in the mandible than in the femur, suggesting that several genes found in this study might be candidates for cementum-specific markers. In this study, a novel experimental system was established to isolate target tissues from undecalcified frozen sections at the cell unit level and to obtain intact RNA. Our methodologies could help to guide further investigation of mineralized tissues and to explore tissue-specific factors.

Periosteum has been demonstrated to contain mesenchymal progenitor cells differentiating to osteoblasts, and both bone morphogenetic protein-2 (BMP-2) and vascular endothelial growth factor (VEGF) may play important roles in cell-based approaches to bone regeneration. The purpose of this study was to evaluate the feasibility and efficacy of BMP-2 and/or VEGF on periosteal cell differentiation to osteoblasts in vitro and ectopic bone formation in vivo. Human periosteum-derived cells were transfected with BMP-2, VEGF, BMP-2 + VEGF, or vehicle as a control by non-viral gene transfer and then cultured and implanted to nude mice intramuscularly. Real-time polymerase chain reaction analysis of the culture revealed that transgenes for BMP-2 and BMP-2 + VEGF induced more mRNA expression of alkaline phosphatase, collagen type I, and osteocalcin than VEGF and vehicle treatments; additionally, alizarin red S staining, alkaline phosphatase staining, and alkaline phosphatase activity were significantly higher in the BMP-2 + VEGF transgene than in the other versions. After implantation, ectopic bone was observed at 4 weeks and greatly increased at 8 weeks in all groups. In particular, the combination of BMP-2 and VEGF formed significantly more bone at 4 weeks, and VEGF transfection resulted in more blood vessels relative to the conditions without VEGF. Thus, VEGF might enhance BMP2-induced bone formation through modulation of angiogenesis. Keywords:: tissue engineering, periosteal cell, bone morphogenetic protein-2 (BMP-2), vascular endothelial growth factor (VEGF), gene transfer

Purpose We examined safety issues related to the presence of various metallic dental materials in magnetic resonance (MR) imaging at 7 tesla. Methods A 7T MR imaging scanner was used to examine 18 kinds of materials, including 8 metals used in dental restorations, 6 osseointegrated dental implants, 2 abutments for dental implants, and 2 magnetic attachment keepers. We assessed translational attraction forces between the static magnetic field and materials via deflection angles read on a tailor-made instrument and compared with those at 3T. Heating effects from radiofrequency during image acquisitions using 6 different sequences were examined by measuring associated temperature changes in agarose-gel phantoms with a fiber-optic thermometer. Results Deflection angles of the metallic dental materials were significantly larger at 7T than 3T. Among full metal crowns (FMCs), deflection angles were 18.0° for cobalt-chromium (Co-Cr) alloys, 13.5° for nickel-chromium (Ni-Cr) alloys, and 0° for other materials. Deflection angles of the dental implants and abutments were minimal, ranging from 5.0 to 6.5°, whereas the magnetic attachment keepers were strongly attracted to the field, having deflection angles of 90° or more. Increases in temperature of the FMCs were significant but less than 1°C in every sequence. The dental implant of 50-mm length showed significant but mild temperature increases (up to 1.5°C) when compared with other dental implants and abutments, particularly on sequences with high specific absorption rate values. Conclusion Although most metallic dental materials showed no apparent translational attraction or heating at 7T, substantial attraction forces on the magnetic attachment keepers suggested potential risks to patients and research participants undergoing MR imaging examinations.

Reduction of loading force to bone induces osteopenia. Although the tail suspension model is the frequently used osteopenia model, this model burdens the animals with nonphysiologic blood distribution and systemic stress. We developed a new simple animal model for osteopenia under reduced loading. Both hind legs of male ICR mice (8 weeks old) in the experimental group were inserted into plastic tubes, which then were connected with wires of three sizes. This apparatus completely immobilized both femurs while allowing the tibias to move. Animals were pair-fed and sacrificed on Days 3, 7, 10, and 14 after immobilization. Bone mineral density measurement with dual energy xray absorptiometry revealed bone loss in the immobilized femurs. Histomorphometric analysis showed increased bone resorption and decreased bone formation, which started from Day 7 and continued until Day 14, resulting in structural disorders in the cancellous bone. Osteoclast population increase before osteoblast population decrease revealed that osteoclasts initially affect the process of this type of osteopenia. Our immobilization model is simple, easy to use, well-tolerated by the animal, and has potential for evaluating the therapeutic effects of drugs to treat osteopenia caused by reduced loading.

INTRODUCTION: Although previous studies have shown that maxillary molar extraction in ovariectomized (OVX) animals causes mandibular loss of bone, it is still questionable as to whether estrogen deficiency affects mandibles with functional occlusion. MATERIALS and METHODS: To answer this question, 13-week-old female Sprague‐Dawley rats were bilaterally OVX or sham-operated. After 109 days, the bone mineral density (BMD) of the femurs and mandibles was measured using dual-energy X-ray absorptiometry (DEXA) and peripheral quantitative computed tomography (pQCT). RESULTS: In DEXA analysis, although the BMD of the total mandible of the OVX rats was similar to that of the sham-operated rats, the BMD of the condylar region in the OVX rats had decreased by 12.5%. In pQCT analysis, decrease in trabecular BMD of the mandibular bone was detectable but low in the molar region (maximal 13%), whereas no difference was seen in cortical BMD. In the femurs, the trabecular bone prominently decreased in OVX rats (30% decrease in pQCT analysis) as previously reported. CONCLUSION: This study revealed regional differences in the mandibular bone decrease in OVX rats. Although the mechanism of low susceptibility of the mandible to estrogen-deficient conditions remains unknown, it is likely that mechanical stress derived from functional occlusion is preventing bone loss in this pathological condition. Furthermore, this study demonstrated the advantage of pQCT in analyzing rat mandibular bone. Oral Diseases (2003) 9, 24–28

Some dental implants are coated with hydroxyapatite (HA), which preferentially binds to bone. Several matrix proteins have an arginine-glycine-aspartic acid (RGD) sequence where cells attach via an integrin receptor. We hypothesized that coating an HA surface with an RGD-containing peptide might enhance the attachment and differentiation of osteoblasts. The HA disks (diameter 34 mm, thickness 1 mm) were treated with a solution (50 mM Tris/HCl and 150 mM NaCl, pH 7.4) containing the peptide EEEEEEEPRGDT, in which the E repetition exerts a high affinity to HA. After washing with phosphate-buffered saline, KUSA/A1 mouse osteoblastic cells were inoculated onto the HA surface and cultured. After 30 min, the number of cells attached to the surface was counted. The DNA content and alkaline phosphatase (ALP) activity were measured after 10 days in culture. Expression of bone matrix proteins was also examined by means of reverse transcriptase-polymerase chain reaction at 7 days; the mineralized area of the culture was also evaluated by staining with Alizarin Red S after 10 days. Treatment with the peptide stimulated cell attachment and increased DNA content and ALP activity. Furthermore, matrix protein expression and mineralized nodule formation were enhanced to a greater extent on the peptide-treated surface than on the nontreated surface. Our results indicate that coating an HA surface with RGD-containing peptide enhances osteoblast attachment and differentiation. This peptide treatment of HA-coated implants may stimulate the osseointegration of the implants.

One G protein-coupled receptor (GPCR) can activate more than one G protein, but the physiologic importance of such activation has not been demonstrated in vivo. We have generated mice expressing exclusively a mutant form of the PTH/PTHrP receptor (DSEL) that activates adenylyl cyclase normally but not phospholipase C (PLC). DSEL mutant mice exhibit abnormalities in embryonic endochondral bone development, including delayed ossification and increased chondrocyte proliferation. Analysis of the differentiation of embryonic metatarsals in vitro shows that PTH(1–34) and forskolin inhibit, whereas active phorbol ester stimulates, hypertrophic differentiation. Thus, PLC signaling via the PTH/PTHrP receptor normally slows the proliferation and hastens the differentiation of chondrocytes, actions that oppose the dominant effects of PTH/PTHrP receptors and that involve cAMP-dependent signaling pathways.

This position paper reviews physical and mechanical properties of thermoplastic resin used for non-metal clasp dentures, and describes feature of each thermoplastic resin in clinical application of non-metal clasp dentures and complications based on clinical experience of expert panels. Since products of thermoplastic resin have great variability in physical and mechanical properties, clinicians should utilize them with careful consideration of the specific properties of each product. In general, thermoplastic resin has lower color-stability and higher risk for fracture than polymethyl methacrylate. Additionally, the surface of thermoplastic resin becomes roughened more easily than polymethyl methacrylate. Studies related to material properties of thermoplastic resin, treatment efficacy and follow-up are insufficient to provide definitive conclusions at this time. Therefore, this position paper should be revised based on future studies and a clinical guideline should be provided.

Summary Prosthetic treatment with dentures is often required for the elderly who have reduced swallowing function. Therefore, it is important to understand the relationship between denture-wearing and feeding function from the perspective of swallowing. To clarify changes in bolus transport during feeding in elderly edentulous patients with or without complete dentures. Subjects were 15 elderly edentulous volunteers who were treated with maxillary and mandibular complete dentures. The test food was 10 g of minced agar jelly containing barium sulphate with a particle diameter of 4·0–5·6 mm. Lateral videofluoroscopy was performed to assess the position of the leading edge of the bolus, the bolus volume in each area at swallow onset, bolus transit time and the mandibular position during pharyngeal swallowing. There were significant changes between the bolus transport with and without dentures. Without dentures, the leading edge of the bolus at swallow onset fell from the valleculae area to the hypopharynx, and the bolus volume in the hypopharynx increased. Bolus transit time increased in the oral cavity, valleculae and hypopharynx. The mandibular position shifted anterosuperior direction. The results arose owing to anatomical changes in the oral and pharyngeal structure and the following functional changes: poor food manipulation, poor bolus formation and delayed swallowing reflex. Removing dentures in elderly edentulous individuals influences bolus transport during feeding, resulting in the exacerbation of the reduced swallowing reserve capacity that accompanies ageing, and may increase the risk of dysphagia.

Transforming growth factor-β (TGF-β) is a critical regulator of osteogenic differentiation and the platelet-derived growth factor (PDGF) is a chemoattractant or mitogen of osteogenic mesenchymal cells. However, the combined effects of these regulators on the osteogenic differentiation of mesenchymal cells remains unknown. In this study, we investigated the effects of TGF-β and/or PDGF on the osteogenic differentiation of human mesenchymal stem cells (hMSCs). The TGF-β-induced osteogenic differentiation of UE7T-13 cells, a bone marrow-derived hMSC line, was markedly enhanced by PDGF, although PDGF alone did not induce differentiation. TGF-β induced extracellular signal-regulated kinase (ERK) phosphorylation and PDGF induced Akt phosphorylation. In addition, the mitogen-activated protein kinase (MAPK)/ERK kinase (MEK) inhibitor, U0126, suppressed the osteogenic differentiation induced by TGF-β alone. Moreover, U0126 completely suppressed the osteogenic differentiation synergistically induced by TGF-β and PDGF, whereas the phosphoinositide-3-kinase (PI3K) inhibitor, LY294002, only partially suppressed this effect. These results suggest that the enhancement of TGF-β-induced osteogenic differentiation by PDGF-induced PI3K/Akt-mediated signaling depends on TGF-β-induced MEK activity. Thus, PDGF positively modulates the TGF-β-induced osteogenic differentiation of hMSCs through synergistic crosstalk between MEK- and PI3K/Akt-mediated signaling.

Denture adhesives, characterized as medical products in 1935 by the American Dental Association, have been considered useful adjuncts for improving denture retention and stability. However, many dentists in Japan are hesitant to acknowledge denture adhesives in daily practice because of the stereotype that dentures should be inherently stable, without the aid of adhesives. The aim of this study is to verify the efficacy of denture adhesives to establish guidelines for Japanese users. The null hypothesis is that the application of denture adhesives, including the cream and powder types, or a control (isotonic sodium chloride solution) would not produce different outcomes nor would they differentially improve the set outcomes between baseline and day 4 post-application. This ten-center, randomized controlled trial with parallel groups is ongoing. Three hundred edentulous patients with complete dentures will be allocated to three groups (cream-type adhesive, powder-type adhesive, and control groups). The participants will wear their dentures with the denture adhesive for 4 days, including during eight meals (three breakfasts, two lunches, and three dinners). The baseline measurements and final measurements for the denture adhesives will be performed on the first day and after breakfast on the fourth day. The primary outcome is a general satisfaction rating for the denture. The secondary outcomes are denture satisfaction ratings for various denture functions, occlusal bite force, resistance to dislodgement, masticatory performance, perceived chewing ability, and oral health-related quality of life. Between-subjects comparisons among the three groups and within-subjects comparisons of the pre- and post-intervention measurements will be performed. Furthermore, a multiple regression analysis will be performed. The main analyses will be based on the intention-to-treat principle. A sample size of 100 subjects per group, including an assumed dropout rate of 10 %, will be required to achieve 80 % power with a 5 % alpha level. This randomized clinical trial will provide information about denture adhesives to complete denture wearers, prosthodontic educators, and dentists in Japan. We believe this new evidence on denture adhesive use from Japan will aid dentists in their daily practice even in other countries. ClinicalTrials.gov NCT01712802 . Registered on 17 October 2012.

The aim of this study was to produce epithelial attachment on a typical implant abutment surface of smooth titanium. A challenging complication that hinders the success of dental implants is peri-implantitis. A common cause of peri-implantitis may results from the lack of epithelial sealing at the peri-implant collar. Histologically, epithelial sealing is recognized as the attachment of the basement membrane (BM). BM-attachment is promoted by activated platelet aggregates at surgical wound sites. On the other hand, platelets did not aggregate on smooth titanium, the surface typical of the implant abutment. We then hypothesized that epithelial BM-attachment was produced when titanium surface was modified to allow platelet aggregation. Titanium surfaces were coated with a protease activated receptor 4-activating peptide (PAR4-AP). PAR4-AP coating yielded rapid aggregation of platelets on the titanium surface. Platelet aggregates released robust amount of epithelial chemoattractants (IGF-I, TGF-β) and growth factors (EGF, VEGF) on the titanium surface. Human gingival epithelial cells, when they were co-cultured on the platelet aggregates, successfully attached to the PAR4-AP coated titanium surface with spread laminin5 positive BM and consecutive staining of the epithelial tight junction component ZO1, indicating the formation of complete epithelial sheet. These in-vitro results indicate the establishment of epithelial BM-attachment to the titanium surface.