Quantitative analysis of change in bone volume 5 years after sinus floor elevation using plate-shaped bone substitutes: a prospective observational study.

PURPOSE: Tricalcium phosphate (TCP) has osteoconductive ability and reportedly offers similar clinical results as autogenous bone grafts in dental implant treatment. However, few reports quantify temporal changes in augmented bone volume after sinus augmentation. We aimed to establish a three-dimensional (3D) quantification method to assess bone volume after sinus augmentation and to evaluate biocompatibility of the TCP plate. METHODS: Maxillary sinus floor augmentation was performed employing the lateral window technique, and plate-shaped ß-TCP (TCP plate) was used instead of granular bone grafting materials. After lifting the sinus membrane, the TCP plate was inserted and supported by dental implants or micro-screws. The changes in bone volumes in the maxillary sinus before and after surgery were recorded using cone-beam computed tomography, saved as Digital Imaging and Communications in Medicine-formatted files, and transformed to Standard Triangle Language (STL)-formatted files. Pre- and post-operative STL data of bone volume were superimposed, and the augmented bone volume was calculated. Moreover, changes in bone volumes, TCP plate resorption rates, and bone heights surrounding the implants were three dimensionally quantified. RESULTS: Fifteen implants in nine subjects were included in this study. TCP plates secured long-term space making, with results similar to those of granular bone substitutes. Newly formed bone was identified around the implant without bone graft material. TCP plate was absorbed and gradually disappeared. CONCLUSIONS: A novel 3D quantification method was established to evaluate changes in bone volume. Clinical application of TCP plate in sinus augmentation could be a better procedure in terms of prognosis and safety.

Self-Prepared Hyaluronic Acid/Alkaline Gelatin Composite with Nano-Hydroxyapatite and Bone Morphogenetic Protein for Cranial Bone Formation.

New bone-forming substitute materials are highly useful in dental implantology. The purpose of this study was to prepare cross-linked hyaluronic acid (cHLA)/cross-linked alkaline gelatin (cAG)/nano-hydroxyapatite (nHAp)/bone morphogenic protein (BMP) constructs; and evaluate their bone-forming capabilities in rat cranial bone defects. The cHLA and cAG liquids processed with an epoxy cross-linker were blended with a 3:1 volume ratio, followed by freeze-drying. The dry composites were further infiltrated with water containing nHAp only (BMP (−)) or with water containing nHAp and BMP (BMP (+)). Prepared wet constructs (BMP (−) and BMP (+)) were implanted in rat cranial bone defects, while defects only were also made, and animals were fed for 8 weeks, followed by subsequent soft X-ray measurements and histological observations. The X-ray results showed that BMP (+) constructs disappeared, though caused inward extension of peripherical bone from defect edges with an increase in length of approximately 24%, larger than those of BMP (−) constructs and defect only with approximately 17% and 8% increments, respectively (p < 0.05). Histological observations of BMP (+) construct samples clearly indicated active bone extension consisting of an array of island-like bones. It was concluded that cHLA/cAG/nHAp/BMP could be used as novel bone-substitute materials.

Bone Formation on Murine Cranial Bone by Injectable Cross-Linked Hyaluronic Acid Containing Nano-Hydroxyapatite and Bone Morphogenetic Protein.

New injection-type bone-forming materials are desired in dental implantology. In this study, we added nano-hydroxyapatite (nHAp) and bone morphogenetic protein (BMP) to cross-linkable thiol-modified hyaluronic acid (tHyA) and evaluated its usefulness as an osteoinductive injectable material using an animal model. The sol (ux-tHyA) was changed to a gel (x-tHyA) by mixing with a cross-linker. We prepared two sol−gel (SG) material series, that is, x-tHyA + BMP with and without nHAp (SG I) and x-tHyA + nHAp with and without BMP (SG II). SG I materials in the sol stage were injected into the cranial subcutaneous connective tissues of mice, followed by in vivo gelation, while SG II materials gelled in Teflon rings were surgically placed directly on the cranial bones of rats. The animals were sacrificed 8 weeks after implantation, followed by X-ray analysis and histological examination. The results revealed that bone formation occurred at a high rate (>70%), mainly as ectopic bone in the SG I tests in mouse cranial connective tissues, and largely as bone augmentation in rat cranial bones in the SG II experiments when x-tHyA contained both nHAp and BMP. The prepared x-tHyA + nHAp + BMP SG material can be used as an injection-type osteoinductive bone-forming material. Sub-periosteum injection was expected.

Preparation of Collagen/Hydroxyapatite Composites Using the Alternate Immersion Method and Evaluation of the Cranial Bone-Forming Capability of Composites Complexed with Acidic Gelatin and b-FGF.

Bone-substitute materials are essential in dental implantology. We prepared collagen (Col)/hydroxyapatite (Hap)/acidic gelatin (AG)/basic fibroblast growth factor (b-FGF) constructs with enhanced bone-forming capability. The Col/Hap apatite composites were prepared by immersing Col sponges alternately in calcium and phosphate ion solutions five times, for 20 and 60 min, respectively. Then, the sponges were heated to 56 °C for 48 h. Scanning electron microscopy/energy-dispersive X-ray spectroscopy, Fourier-transform infrared spectroscopy, and X-ray diffraction analyses showed that the Col/Hap composites contained poorly crystalline Hap precipitates on the Col matrix. Col/Hap composite granules were infiltrated by AG, freeze-dried, and immersed in b-FGF solution. The wet quaternary constructs were implanted in rat cranial bone defects for 8 weeks, followed by soft X-ray measurements and histological analysis. Animal studies have shown that the constructs moderately increase bone formation in cranial bone defects. We found that an alternate immersion time of 20 min led to the greatest bone formation (p < 0.05). Constructs placed inside defects slightly extend the preexisting bone from the defect edges and lead to the formation of small island-like bones inside the defect, followed by disappearance of the constructs. The combined use of Col, Hap, AG, and b-FGF might bring about novel bone-forming biomaterials.

Current complications and issues of implant superstructure.

The purpose of this review is to search for complications of dental implant superstructures and consider the issues involved. This narrative review was performed by searching through PubMed databases and review articles that were published after 1990. Misfitting of the superstructure can result in loosening of screws, reduced preload, and in some cases, significant stress around the implant. External connection modalities and single implant prostheses have been reported to have more loose or broken abutment screws. In addition, when zirconia abutment was used for platform shifting, the rate of fracture of the abutment was considered to be high. Additionally, it was reported that men were significantly at an increased risk of abutment fracture. As for the retention mechanism of implant overdenture, stud attachment (Locator type) should receive more attention to wear and damage of retention parts than other attachments. The causes of the complications of implant superstructures have not been clarified in some cases, and further verification is required. Verification of complications is considered important to obtain a long-term prognosis for superstructures of implants. It will be necessary to further verify complications of implants in the future.

Bone Regeneration of Critical-Size Calvarial Defects in Rats Using Highly Pressed Nano-Apatite/Collagen Composites.

Osteo-conductive bone substitute materials are required in dentistry. In this study, highly pressed nano-hydroxyapatite/collagen (P-nHAP/COL) composites were formed by a hydraulic press. Critical-size bone defects (Φ = 6 mm) were made in the cranial bones of 10-week-old Wistar rats, in which P-nHAP/COL and pressed collagen (P-COL) specimens were implanted. Defect-only samples (DEF) were also prepared. After the rats had been nourished for 3 days, 4 weeks, or 8 weeks, ossification of the cranial defects of the rats was evaluated by micro-computed tomography (micro-CT) (n = 6 each). Animals were sacrificed at 8 weeks, followed by histological examination. On micro-CT, the opacity of the defect significantly increased with time after P-nHAP/COL implantation (between 3 days and 8 weeks, p < 0.05) due to active bone regeneration. In contrast, with P-COL and DEF, the opacity increased only slightly with time after implantation, indicating sluggish bone regeneration. Histological inspections of the defect zone implanted with P-nHAP/COL indicated the adherence of multinucleated giant cells (osteoclasts) to the implant with phagocytosis and fragmentation of P-nHAP/COL, whereas active bone formation occurred nearby. Fluorescent double staining indicated dynamic bone-formation activities. P-nHAP/COL is strongly osteo-conductive and could serve as a useful novel bone substitute material for future dental implant treatments.

Characterization of Five Collagenous Biomaterials by SEM Observations, TG-DTA, Collagenase Dissolution Tests and Subcutaneous Implantation Tests.

Collagenous biomaterials that are clinically applied in dentistry have dermis-type and membrane-type, both of which are materials for promoting bone and soft tissue formation. The properties of materials supplied with different types could affect their biodegradation periods. The purpose of this study was to characterize five of these products by four different methods: scanning electron microscopy (SEM) observation, thermogravimetry-differential thermal analysis (TG-DTA), 0.01 wt% collagenase dissolution test, and subcutaneous implantation test in vivo. SEM micrographs revealed that both dermis and membranous materials were fibrous and porous. The membranous materials had higher specific derivative thermal gravimetry (DTG) peak temperatures in TG-DTA at around 320 °C, longer collagenase dissolution time ranging from about 300 to 500 min, and more longevity in mice exceeding 9 weeks than the dermis materials. There existed a correlation between the peak temperature in TG-DTA and the collagenase dissolution time. It was considered that higher cross-link degree among collagen fibrils of the membrane-type collagenous materials might account for these phenomena. The experimental protocol and numerical results obtained could be helpful for selection and future development of fibrous collagenous biomaterials in clinical use.

A review of the recent literature on maxillary overdenture with dental implants.

The recent literature on maxillary implant overdenture (IOD) was reviewed in order to clarify its predictability and establish treatment guidelines. Electronic searches were performed using PubMed, and articles about maxillary IOD written after 1990 were reviewed, focusing on the following items: I. implant survival rate, II. maxillary IOD survival rate, III. number of implants, IV. attachment type, V. follow-up period, VI. implant system, and VII. opposing dentition. The review revealed an implant survival rate of 61-100% and an overdenture survival rate of 72.4-100%. The attachments used included bars, balls, locators, and telescope crowns. The minimum and maximum observation periods were 12 months and 120 months, respectively, and the number of implants used for supporting IOD ranged from 2 to 8. At present, there is no strong evidence to indicate that maxillary IOD is clearly superior for all the items examined. However, the existing data indicate that maxillary IOD has almost the same therapeutic effect as fixed implant superstructures, and is a treatment option that can be actively adopted for patients in whom fixed superstructures cannot be applied for various reasons.

Applications of three-dimensional printers in prosthetic dentistry.

This narrative review aims to provide an overview of recent studies and case reports on three-dimensional (3D) printing, and to verify the applicability of 3D printers in the field of dental prostheses. This review was performed by conducting a search of PubMed. The clinical application of fabricating a prosthesis made with cobalt-chromium is considered possible depending on the material and hardware of the 3D printer. However, it is currently difficult to assess the clinical use of 3D-printed zirconia crowns. Further research is required, such as verification of materials used, margin morphology, and hardware. Clinically acceptable results have been reported for patterns using 3D printers. Interim restorations made using a 3D printer have been reported with good results that are considered clinically usable. Dentures made with 3D printers need further verification in terms of strength and deformation. Custom trays made with 3D printers are clinically useful, however, issues remain with design time and effort. Although several studies have reported the usefulness of 3D printers, further verification is required since 3D printers are still considered new technology.

Titanium nanoparticles potentially affect gingival tissue through IL-13α2 receptor expression.

PURPOSE: To determine the effects of titanium nanoparticles, that may have been scattered after dental implant placement, on gene and promoter expression, and gingival tissue. METHODS: Ca9-22 cell lines were used as gingival epithelial cells to assess the effects of titanium dioxide nanomaterials as titanium nanoparticles. Cells were cocultured with or without titanium dioxide nanomaterials prior to gene and promoter expression analysis. Expression of interleukin-13α2 receptor was investigated using real-time quantitative reverse-transcription polymerase chain reaction and immunofluorescence staining. Additionally, the enhanced messenger ribonucleic acid (mRNA) expression of transforming growth factor ß1 was analyzed using the same method. RESULTS: Titanium dioxide nanomaterials affected gene and promoter expression in Ca9-22 cells: among the 160 upregulated genes, the upregulation of IL13RA2, which encodes interleukin-13α2 receptor, was the highest (8.625 log2 fold change). Immunofluorescence staining confirmed the increased expression of interleukin-13α2 receptor, which enhanced transforming growth factor ß1 expression by stimulation with interleukin-13. CONCLUSION: Titanium dioxide nanomaterials applied on the gingival epithelium around the dental implant may increase interleukin-13α2 receptor expression. In turn, this can enhance the secretion of transforming growth factor ß1, which is known to promote the differentiation of osteoclasts involved in bone resorption, and potentially affect gingival tissue.

Sinus floor elevation using tricalcium phosphate plate.

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.

Accuracy and practicality of intraoral scanner in dentistry: A literature review.

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.

Establishment of perpendicular protrusion of type I collagen on TiO2 nanotube surface as a priming site of peri-implant connective fibers.

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 < 0.01, a 1-way ANOVA and Tukey HSD post hoc test). Furthermore, TNT-CoIEPF surface rejected epithelial cell stretching and epithelial sheet formation. Chemically linked horizontal CoI on titanium oxide (TiO2) facilitated epithelial cell stretching and sheet formation.

Retrograde transport of masseter muscle-derived neprilysin to hippocampus.

Although the effects of neprilysin (NEP), also called CD10, on the clearance of Alzheimer's disease (AD)-associated amyloid-ß (Aß) 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.

A comparison of diagnosis of early stage interproximal caries with bitewing radiographs and periapical images using consensus reference.

OBJECTIVES:: The purpose of this study was to assess the diagnostic property of intraoral bitewing radiographs (BTW) for early stage interproximal caries, and to compare them with periapical radiographs (PA) at different levels of caries progression. METHODS:: A total of 241 interproximal surfaces of BTW and corresponding PAs were used. Seven teaching faculty consisting of three oral and maxillofacial radiologists, two operative faculty, and two prosthodontists evaluated the images. The observers graded images as either "intact", "enamel caries <1/2 width", "enamel caries >1/2 width", or "caries into dentin". The gold-standard was established by consensus of two experienced faculty with 35 years and 27 years of experience. Specificity, sensitivity, positive-predictive value, and negative-predictive value were calculated for the different level of caries progression. Furthermore, receiver operating curves) of BTW and PAs of each evaluator were made and the area under the curve of BTW and PAs were compared. RESULTS:: There was no significant difference in the specificity of BTW and PAs. BTW showed significantly higher sensitivity than PAs in all levels of caries progression (p<0.01). Positive-predictive value and negative-predictive value of BTWs were also significantly higher than PAs. One-way ANOVA and Tukey HSD test showed a significant difference in sensitivity with different levels of caries progression. The average area under the curve was significantly higher for BTWs than PAs (p<0.01). CONCLUSIONS:: BTWs offer a significant advantage over PAs in the diagnoses of early stages of interproximal carious lesions.

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.

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 0.5­1.0 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 ß-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.