Hidemichi Kihara

Effects of the combination with α-tricalcium phosphate and simvastatin on bone regeneration

BACKGROUND Although local application of statins stimulates bone formation, high dose of simvastatin induces inflammation. OBJECTIVE A study was conducted to test the hypothesis that maximum bone regeneration with less inflammation would be achieved by combining an optimal dose of simvastatin with alpha-tricalcium phosphate (alpha-TCP), which is an osteoconductive biomaterial capable of releasing the drug gradually. MATERIAL AND METHODS Bilateral 5-mm-diameter calvarial defects were created in adult Wistar rats and filled with preparations of different doses of simvastatin (0, 0.01, 0.1, 0.25 and 0.5 mg) combined with alpha-TCP particles or left empty. The animals were sacrificed at 2, 4 and 8 weeks and analyzed radiologically and histologically. Half of the animals of 4 and 8 weeks were labeled with fluorescence dyes and histomorphometrically analyzed. RESULTS Simvastatin doses of 0.25 and 0.5 mg caused inflammation of the soft tissue at the graft site whereas control and other doses did not. The micro-CT analysis revealed that the alpha-TCP with 0.1 mg simvastatin (TCP-0.1) group yielded significantly higher bone volumes than untreated control group at all three time points (249%, 227% and 266% at 2, 4 and 8 weeks, respectively). The percentage of defect closure, bone mineral content and bone mineral density were also higher in the TCP-0.1 group than in the other groups. CONCLUSION When combined with alpha-TCP particles, 0.1 mg simvastatin is the optimal dose for stimulation of the maximum bone regeneration in rat calvarial defects without inducing inflammation and it could be applied as an effective bone graft material.

Effects of apatite particle size in two apatite/collagen composites on the osteogenic differentiation profile of osteoblastic cells.

The development of new osteoconductive bone substitute materials is expected in medicine. In this study, we attempted to produce new hydroxylapatite (HAP)/collagen (Col) composites using two HAP particles of different sizes and porcine type I collagen. The two HAP particles were either nano-sized (40 nm in average diameter; n-HAP) or had macro-pore sizes of 0.5–1.0 mm in length with fully interconnected pores (m-HAP). The aim of this study was to investigate the effects of apatite particle size in two HAP/Col composites on the osteogenic differentiation profile in osteoblast-like cells (SaOS-2). We created a collagen control sponge (Col) and two HAP/Col composite sponges (n-HAP/Col and m-HAP/Col) using freeze-drying and dehydrothermal cross-linking techniques, and then punched out samples of 6 mm in diameter and 1 mm in height. The SaOS-2 cells were cultured on three test materials for 1, 2, 3 and 4 weeks. Total RNA was extracted from the cultured cells and the expression of osteogenic differentiation-related genes was evaluated by reverse transcription PCR (RT-PCR) using primer sets of alkaline phosphatase (ALP), type 1 collagen (COL1), bone sialoprotein (BSP) and osteocalcin precursor [bone gamma-carboxyglutamate (gla) protein (BGLAP)] genes, as well as the β-actin gene. The cells were also cultured on Col, n-HAP/Col and m-HAP/Col specimens for 1 and 4 weeks, and were then observed under a scanning electron microscope (SEM). The experimental results were as follows: RT-PCR indicated that osteogenic differentiation, particularly the gene expression of BSP, was most accelerated when the cells were cultured on n-HAP/Col specimens, followed by m-HAP/Col, whilst the weakest accelaeration was observed when the cells were cultured on Col specimens. As shown by the SEM images, the SaOS-2 cells were fibroblastic when cultured on Col specimens for up to 4 weeks; they were fibroblastic when cultured on n-HAP/Col specimens for 1 week, but appeared as spheroids, while actively phagocytizing n-HAP particles at 4 weeks; however, they appeared as deformed fibroblasts when cultured on m-HAP/Col specimens, detached from the particles. Despite limited experimental results, our study suggests that n-HAP/Col may be employed as a new osteoconductive bone substitute material.

Examination of the Position Accuracy of Implant Abutments Reproduced by Intra-Oral Optical Impression

An impression technique called optical impression using intraoral scanner has attracted attention in digital dentistry. This study aimed to evaluate the accuracy of the optical impression, comparing a virtual model reproduced by an intraoral scanner to a working cast made by conventional silicone impression technique. Two implants were placed on a master model. Working casts made of plaster were fabricated from the master model by silicone impression. The distance between the ball abutments and the angulation between the healing abutments of 5 mm and 7 mm height at master model were measured using Computer Numerical Control Coordinate Measuring Machine (CNCCMM) as control. Working casts were then measured using CNCCMM, and virtual models via stereo lithography data of master model were measured by a three-dimensional analyzing software. The distance between ball abutments of the master model was 9634.9 ± 1.2 μm. The mean values of trueness of the Lava COS and working casts were 64.5 μm and 22.5 μm, respectively, greater than that of control. The mean of precision values of the Lava COS and working casts were 15.6 μm and 13.5 μm, respectively. In the case of a 5-mm-height healing abutment, mean angulation error of the Lava COS was greater than that of the working cast, resulting in significant differences in trueness and precision. However, in the case of a 7-mm-height abutment, mean angulation errors of the Lava COS and the working cast were not significantly different in trueness and precision. Therefore, distance errors of the optical impression were slightly greater than those of conventional impression. Moreover, the trueness and precision of angulation error could be improved in the optical impression using longer healing abutments. In the near future, the development of information technology could enable improvement in the accuracy of the optical impression with intraoral scanners.

Impact of the Static and Radiofrequency Magnetic Fields Produced by a 7T MR Imager on Metallic Dental Materials

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.

Function of Platelet-Induced Epithelial Attachment at Titanium Surfaces Inhibits Microbial Colonization:

The aim of this study was to evaluate the barrier function of platelet-induced epithelial sheets on titanium surfaces. The lack of functional peri-implant epithelial sealing with basal lamina (BL) attachment at the interface of the implant and the adjacent epithelium allows for bacterial invasion, which may lead to peri-implantitis. Although various approaches have been reported to combat bacterial infection by surface modifications to titanium, none of these have been successful in a clinical application. In our previous study, surface modification with protease-activated receptor 4–activating peptide (PAR4-AP), which induced platelet activation and aggregation, was successful in demonstrating epithelial attachment via BL and epithelial sheet formation on the titanium surface. We hypothesized that the platelet-induced epithelial sheet on PAR4-AP–modified titanium surfaces would reduce bacterial attachment, penetration, and invasion. Titanium surface was modified with PAR4-AP and incubated with platelet-rich plasma (PRP). The aggregated platelets released collagen IV, a critical BL component, onto the PAR4-AP–modified titanium surface. Then, human gingival epithelial cells were seeded on the modified titanium surface and formed epithelial sheets. Green fluorescent protein (GFP)–expressing Escherichia coli was cultured onto PAR4-AP–modified titanium with and without epithelial sheet formation. While Escherichia coli accumulated densely onto the PAR4-AP titanium lacking epithelial sheet, few Escherichia coli were observed on the epithelial sheet on the PAR4-AP surface. No bacterial invasion into the interface of the epithelial sheet and the titanium surface was observed. These in vitro results indicate the efficacy of a platelet-induced epithelial barrier that functions to prevent bacterial attachment, penetration, and invasion on PAR4-AP–modified titanium.

A Survey of Dental Implant Instruction in Predoctoral Dental Curricula in North America

The aim of this study was to investigate the extent and forms of predoctoral implant dentistry instruction in North American dental schools and to identify future directions and challenges. The survey collected data on curriculum content, departmental oversight, techniques, and materials, as well as current problems to be solved. The 30-question survey was sent in 2012 to the dean or administrator in charge of the predoctoral curriculum of all 73 dental schools in the U.S. and Canada at the time; four reminders were sent. Forty-seven schools responded, for a response rate of 64%. Of the 47 responding schools, 46 (98%) offered didactic instruction (mean of 17 hours); 87% had a laboratory component (mean of 14.46 hours); and 57% had a clinical requirement. In the responding schools, students had an average of 1.85 implant restorative cases and 0.61 surgical cases. Forty-two of the schools (89%) had implemented observation of implant surgery and/or assisting with implant surgery in their curricula. Major challenges reported in implementing a comprehensive predoctoral implant curriculum included expense of implant systems to the schools and to patients, shortage of predoctoral cases, and lack of curriculum time and trained faculty. These results show that implant education for predoctoral dental students continues to expand, with a trend towards more preclinical exercises and clinical experiences and fewer didactic courses.

Knowledge and Practice of Oral Cancer Screening in Teaching Faculty—Comparison of Specialty and Year of Clinical Experience

The purpose of this study was to assess the knowledge, practice, confidence, and perceived barriers to oral cancer screening among teaching faculty in Japan. Results were compared by specialist as well as years of clinical experience. A 25-question survey was used to assess the oral cancer screening practices of faculty dentists at Iwate Medical University, School of Dental Medicine, the only dental school located in the northeast (Tohoku) region of Japan. The study was approved by the Institutional Review Board of Iwate Medical University. The response rate was 83% (n = 110, 71.8% were male). This survey revealed that only 43.6% of the dentists performed oral cancer screening frequently (always or usually) at the initial appointment, and there was no significant differences between specialists and clinical experience. Visual inspection of the oral cavity was the primary screening method, but the frequency and content of the examination (TMJ and tonsil) was significantly different between specialties. A history of cancer and tobacco use motivated providers to perform an examination and was significantly different between various specialists and clinically experienced providers. In contrast, HPV and alcohol consumption were a weak motivator. The confidence level of providers on their examination knowledge/skills was poor, especially among junior faculty. More than 80% of junior faculty indicated a lack of knowledge/skills as a major barrier.

Epithelial cell adhesion efficacy of a novel peptide identified by panning on a smooth titanium surface.

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.