Analysis of Bone Mineral Density and Bone Quality of Cortical Bone in the Human Hyoid Body and Histological Observation of the Entheses.

The hyoid is the only bone in the human body that is completely independent, not forming a joint with any other bone; its position is maintained by the suprahyoid and infrahyoid muscles, as well as several ligaments. The purpose of this study was to ascertain the effect of the functional pressure arising from these muscles and ligaments on the hyoid body structure from its bone mineral density, bone quality, and histological observations. The area between the mesial-most part of each lesser horn and the center of the hyoid body was divided equally into four measurement regions. We conducted histological investigations at each measurement region and observed the entheses. To analyze bone mass and bone quality, we also measured bone mineral density (BMD) and analyzed biological apatite (BAp) crystallite orientation in the same regions. Histological observations identified periosteal insertions and fibrocartilaginous entheses. There was no significant difference in BMD between any of the measurement regions, but the preferential orientation of BAp crystallites was stronger in the infrahyoid muscles and ligaments, where fibrocartilaginous entheses are found, than in other places. This suggests that the functional pressure at these sites might exert a major effect not only on the morphological characteristics of the entheses but also on bone quality.

The Wear Behavior of Glass-Ceramic CAD/CAM Blocks against Bovine Enamel.

The wear of enamel and crown restorative materials often occur by occlusion. The purpose of this study was to evaluate the wear volume between glass-ceramics used for CAD/CAM blocks (lithium disilicate: Initial LiSi block (LIS), IPS e.max CAD (IPS), zirconia-reinforced lithium silicate glass-ceramics: Celtra DUO (DUO), VITA Suprinity (VITS) and feldspar-based glass-ceramics: Vitablocs Mark II (MAK)) and bovine tooth enamel using a two-body wear test, the hardness, three-point bending strength, micro-structure and the element components of glass-ceramics. The data were analyzed using a one-way analysis of variance and Tukey's multiple comparison test (α = 0.05). IPS and DUO with relatively large size crystal gain had significantly larger abrader wear volumes. Zirconia-reinforced lithium silicate glass-ceramics (DUO, VITS) caused significantly greater wear volume in antagonist enamel. MAK with scale-shape crystals grains produced distinct scratches after wear tests, both in the material itself and in the enamel. A strong correlation between the mechanical properties (hardness, three-point bending strength) and wear volume could not be confirmed. The type of glass-ceramic, size, and shape of the crystal grains affected the wear behavior of the glass-ceramics for CAD/CAM blocks. Therefore, dentists should consider that wear behavior varies with crystal structure, size, and shape in glass-ceramics for CAD/CAM blocks.

Effect of the Correction of Bilateral Differences in Masseter Muscle Functional Pressure on the Mandible of Growing Rats.

The objective of this study is to clarify the effect of restoring the lowered masticatory muscle functional pressure and correcting bilateral differences in masticatory muscle functional pressure on jawbone growth during growth and development with a quantitative evaluation of the changes in the micro/nanostructural characteristics of entheses. Male Wistar rats aged 4 weeks were divided into an experimental group injected with a botulinum toxin serotype A (BoNT/A) formulation to reduce muscle function (BTX group) and a control group (CTRL group). They were euthanised after 6, 8, 10, 12, and 16 weeks after measuring the difference between the midline of the upper and lower incisors. The mandibles were harvested for histological examination, second harmonic generation imaging, and the quantitative evaluation of biological apatite (BAp) crystal alignment. The midline difference decreased with age in weeks. In rats from 6 weeks after BoNT/A administration to 12 weeks after administration, the collagen fibre bundle diameter was significantly smaller in the BTX group; the difference between the two groups decreased with increasing age. BAp crystal alignment was significantly different on the x-axis and the y-axis on the BTX group from 6 weeks after BoNT/A administration to 10 weeks after administration. Asymmetry of mandibular bone formation caused by load imbalance during growth could be corrected by the adjustment of the function of the masseter muscle on either side.

Alveolar Bone Microstructure Surrounding Orthodontic Anchor Screws with Plasma Surface Treatment in Rats.

A lateral load was applied to anchor screws that had undergone surface treatment, and the structure, cellular dynamics, and quality of the bone surrounding anchor screws were analyzed to investigate the effect of this surface treatment on the peri-implant jawbone. In addition, bone microstructural characteristics were quantitatively evaluated for each site of loading on the bone around the anchor screw. Rats were euthanized after observation on days 3, 5, or 7, and bone quality analyses were performed. Bone-implant contact rate increased more rapidly at an early stage in the treated surface group than in the untreated surface group. Bone lacuna morphometry showed that the measured values adjacent to the screw at the screw neck on the compressed side (A) and at the screw tip on the uncompressed side (D) were significantly lower than those at the screw tip on the compressed side (B) and at the screw neck on the uncompressed side (C). Collagen fiber bundle diameter showed that the measured values adjacent to regions A and D were significantly higher than those at regions B and C. Anchor screw surface activation facilitates initial bone contact of the screw, suggesting that early loading may be possible in clinical practice.

The effects of orthodontic anchor screw inserted into the femur of growth-phase or mature rats -Osteoid formation, bone mineral density, collagen fiber bundles, biological apatite crystal orientation.

The purpose of this study is to investigate the effect of orthodontic anchor screws (OASs) inserted into the femur of growth-phase or mature rats using histological observation and bone structure analysis. The experimental animals are growth-phase (6-week-old) or mature (25-week-old) male Wistar rats. OAS was placed into the point one-third of the femoral length from the proximal end of the femur, and the response of the surrounding bone was observed and measured. The results showed at the OAS bone interface, in growth-phase rats, bone mineral density (BMD) was reduced and the running angle of collagen fiber bundles varied significantly. In mature rats, more osteoid was observed and biological apatite (BAp) crystals showed a different orientation. It was suggested that after the insertion of OASs, bone volume and quality are decreased, but after a sufficient healing period, a new bone micro/nano structure, different from the original structure, are reconstructed.

Effects of hollow structures added by selective laser sintering on the mechanical properties of Co-Cr alloy.

PURPOSE: This study investigates the effects of hollow structures, added by selective laser sintering (SLS), on the mechanical properties of a Co-Cr alloy for providing an optimal structural property to the framework components of removable partial dentures (RPDs). METHODS: The specimens produced using the 3D data of the dumbbell-shaped cylinders were divided into four groups based on the manufacturing method: Cast, Mill, SLS-solid, and SLS-hollow. Tensile tests were performed to measure the mechanical properties of the specimens. The mechanical property values among the four groups were statistically compared using the Kruskal-Wallis test followed by the Steel-Dwass test (α = 0.05). RESULTS: The median elastic modulus was the largest in the Cast, followed by SLS-solid, Mill, and SLS-hollow, with no significant differences observed between all conditions. The median ultimate tensile strength was the largest in the order of SLS-solid, Mill, SLS-hollow, and Cast. The median 0.2% proof stress was the largest in SLS-solid, followed by SLS-hollow, Cast, and Mill. The median elongation was the highest in the order of Mill, SLS-solid, SLS-hollow, and Cast. CONCLUSIONS: With the addition of hollow structures, the elastic modulus decreased while the mechanical strength and proof stress remained high in SLS specimens. In addition, the ISO 22674 standard for dental metals was met, suggesting that SLS may be a possible method to design RPD frameworks with high strength and optimal structural properties.

Effect of Surface Treatments of Polyetherketoneketone as a Post Material on Shear Bond Strength to Root Dentin using Two Types of Resin Cement.

PURPOSE: This study investigated the effects of mechanical and chemical pretreatment of polyetherketoneketone (PEKK) on shear bond strength (SBS) to root dentin using two types of resin cement. MATERIALS AND METHODS: A total of 100 PEKK specimens were prepared and polished. Sixty specimens were mechanically treated by sandblasting (50-µm Al2O3 for 10 s at 0.2-0.25 MPa), and the remaining 40 were untreated. Self-adhesive resin cement and conventional resin cement were used. PEKK specimens bonded to root dentin using self-adhesive resin cement were classified into three groups by pretreatment method: (1) untreated PEKK, (2) mechanical pretreatment (sandblasted PEKK), and (3) both mechanical and chemical pretreatment (PEKK sandblasting as well as application of Scotchbond Universal; 3M Oral Care). Conventional resin cement was pretreated following the same steps (1-3) as those followed for self-adhesive specimens. Each group included 10 specimens. PEKK specimens after surface treatments were examined using SEM. SBS tests were performed using a universal testing machine, and data were statistically analyzed using one-way ANOVA and Tukey's multiple comparison test (p < 0.05). RESULTS: No significant difference was observed between cements with and without sandblasting. However, self-adhesive specimens with both mechanical and chemical pretreatments demonstrated higher SBS than other cements with or without pretreatment. CONCLUSION: Mechanical pretreatment by sandblasting did not improve the PEKK-root dentin SBS. However, combined mechanical (sandblasting) and chemical pretreatment (ScotchBond Universal) significantly improved the SBS between the PEKK and root dentin.

Effect of bone defect width and a ferrule on the fracture characteristics of vertically fractured teeth reattached with adhesive resin cement: An in vitro study.

This study aimed to clarify the effects of vertical bone defect width and a ferrule on fracture of the fragments of fractured tooth reattached with adhesive resin cement (reattached tooth). The reattached tooth was built up by a fiber post and composite resin core for abutment and formed to the abutment with or without a ferrule. The vertical bone defect was fabricated with a V-shaped defect in different widths. The fracture load was evaluated using a universal testing machine. The vertical bone defect did not affect the fracture load, but a ferrule increased the root fracture load. For the specimens without a ferrule, debonding between the composite resin core and the root at the coronal loading side and fractures at the apical side of the root were found. In conclusion, the ferrule at abutment could affect fracture load and modes, and the bone defect width did not.

Structural characteristics of the bone surrounding dental implants placed into the tail-suspended mice.

BACKGROUND: There are many unclear points regarding local structural characteristics of the bone surrounding the implant reflecting the mechanical environment. PURPOSE: The purpose of this study is to quantitatively evaluate bone quality surrounding implants placed into the femurs of mice in an unloading model, and to determine the influence of the mechanical environment on bone quality. METHODS: Twenty 12-week-old male C57BL6/NcL mice (n = 5/group) were used as experimental animals. The mice were divided into two groups: the experimental group (n = 10) which were reared by tail suspension, and the control group (n = 10) which were reared normally. An implant was placed into the femur of a tail-suspended mouse, and after the healing period, they were sacrificed and the femur was removed. After micro-CT imaging, Villanueva osteochrome bone stain was performed. It was embedded in unsaturated polyester resin. The polymerized block was sliced passing through the center of the implant body. Next, 100-µm-thick polished specimens were prepared with water-resistant abrasive paper. In addition to histological observation, morphometric evaluation of cancellous bone was performed, and the anisotropy of collagen fibers and biological apatite (BAp) crystals was analyzed. RESULTS: As a result, the femoral cortical bone thickness and new peri-implant bone mass showed low values in the tail suspension group. The uniaxial preferential orientation of BAp c-axis in the femoral long axis direction in the non-implant groups, but biaxial preferential orientation of BAp c-axis along the long axis of implant and femoral long axis direction were confirmed in new bone reconstructed by implant placement. Collagen fiber running anisotropy and orientation of BAp c-axis in the bone surrounding the implant were not significantly different due to tail suspension. CONCLUSIONS: From the above results, it was clarified that bone formation occurs surrounding the implant even under extremely low load conditions, and bone microstructure and bone quality adapted to the new mechanical environment are acquired.

Retention force of fiber-reinforced composite resin post on resin composite for core buildup -Effects of fiber orientation, silane treatment and thermal cycling.

The purpose of this study was to investigate the effects of fiber orientation, silane treatment, and thermal cycling on the retention force of fiber-reinforced composite resin (FRC) posts on resin composite. Two types of FRC posts (linear and woven) were prepared with and without silane treatment. Pull-out test specimens were made of FRC posts and resin composite for core buildup. Specimens were either incubated in distilled water for 24 h or subjected to 10,000 thermal cycles. The maximum fracture load obtained by a pull-out test was defined as the retention force. Fracture modes were observed after the test. Fiber orientation and thermal cycling did not affect the retention force on resin composite, and the retention force was improved by silane treatment. Whereas, fiber orientation affected the fracture mode. The result showed woven fiber orientation could contribute to the mechanically interlocking with the resin composite.

Severe Hydroxyapatite Deposition Disease in Binturongs (Arctictis binturong).

In a collection of 6 young binturongs (Arctictis binturong), 3 presented with anorexia, lethargy, and footpad swelling diagnosed by surgical biopsy as calcinosis circumscripta. Despite supportive care over the next 4 years, affected binturongs had progressive weight loss, hyperphosphatemia, and hyperkalemia and developed large radiodense deposits in tissues adjacent to appendicular joints, thoracolumbar vertebrae, and the sternum. Two binturongs died and necropsies showed severe periarticular mineral deposition with fibrosis and granulomatous inflammation. Additionally, binturongs had mineralization of distal renal tubules, mineralization of the renal interstitium, and marked mineralization of the pulmonary interstitium and peribronchiolar smooth muscle. Foot biopsies from the surviving binturong were evaluated by spectroscopic microanalysis and were positive for hydroxyapatite, Ca10(PO4)6(OH)2. Periarticular and footpad mineralization in these binturongs is consistent with hydroxyapatite deposition disease, which is rarely described in humans and animals as a primary familial condition or a secondary condition often associated with renal injury.

Localization of T-cell factor 4 positive fibroblasts and CD206-positive macrophages during skeletal muscle regeneration in mice.

Skeletal muscle regeneration is initiated by the activation of the transcription factor paired box 7 (Pax7), which is expressed in the satellite cells. The nuclear transcription factor T-cell factor 4 (Tcf4) is expressed in the fibroblasts and is involved in muscle tissue repair, while M2-like macrophages play an important role in skeletal muscle regeneration. However, the localization of M2-like macrophages and the expression of Tcf4 over a period of time during skeletal muscle regeneration remain unknown. Therefore, the murine masseter muscle was immunofluorescence investigated for the surface protein CD206 of M2-like macrophages and Tcf4 of fibroblasts during skeletal muscle regeneration to understand the changes in the CD206 and Tcf4 expression over time. We observed that CD206 entered the cytoplasm of some regenerating muscle fibers 5-7 days after the experimental muscle damage, that is, in the early stage of maturation of the regenerating muscle fibers with central nuclei. In addition, Tcf4 was expressed in the nuclei of the fibroblasts around the regenerating muscle fibers and in the central nuclei of the regenerating muscle fibers. Furthermore, the expression of laminin adjacent to Tcf4-positive cells was observed to partially disappear, and the shape of this missing part was observed to be identical to that of the nuclei of Tcf4-positive cells adjacent to the laminin. Clathrin was also expressed in these sites, demonstrating endocytosis. Thus, these results suggest that in the early stage of maturation of the regenerating muscle fibers, M2-like macrophages and Tcf4-positive fibroblasts enter the cytoplasm of the regenerating muscle fibers, thereby regulating the expression of various maturation factors.

Micro- and nano-bone analyses of the human mandible coronoid process and tendon-bone entheses.

The coronoid process provides attachment to temporalis and masseter muscles, and thus plays an important role in mastication. Tendons connect muscles and bones, mediating the transmission of functional loads to bones. Thus, tendon-bone entheses govern mechanical stress in bones. The preferential orientation of biological apatite (BAp) crystallites, the main mineral component in bones, is an important index for bone quality and function, and is largely influenced by locally applied stress. In this study, we analyzed BAp orientation, Young's modulus, and bone mineral density (BMD) at different sites in the human coronoid process. No differences in BMD were found among the analyzed sites, but BAp crystal orientation was observed to differ. BAp crystallites showed a uni-directional orientation in the mesiodistal direction at the coronoid process apex, but were oriented in the direction vertical to the occlusal plane at other sites. Young's modulus tended to vary according to the BAp orientation. At the apex, a tendon form with characteristics different from those at other sites, including the presence of a fibrocartilaginous layer that may act as a stretching brake to control stress concentration, was observed. These findings suggest that the functional pressure of the temporalis muscle affects bone quality and strength.

Effect of Mesenchymal Cells on Myoblast Sheets Embedded in Collagen Gel.

The objective of this study was to investigate the effects of mesenchymal cells on myoblasts in long-term cultivation of myoblast cell sheets. Sheets of myoblasts and mesenchymal cells from Japanese rabbit oral mucosa were generated and analyzed by histochemistry, Western blot, and reverse transcription-polymerase chain reaction. The presence of desmin and type IV collagen, which is seen in normal muscle tissue, was also confirmed in all the sheets produced. Expression of desmin and type IV collagen showed a decrease under co-culture conditions. In addition, expression of genes important in maintaining the undifferentiated state (Pax7, CD34, myogenin, MyoD) in myoblasts was observed throughout the long cultivation period. Insulin-like growth factor was expressed only when the mesenchymal cells were co-cultured with myoblasts. These data suggest that the presence of mesenchymal cells in a long-term co-culture system influences myoblast differentiation.

Anatomic and Histological Study of Lingual Nerve and Its Clinical Implications.

Although the risk of injuring the lingual nerve in the mandibular molar area during dental treatment is high, it can be repaired by nerve grafting. However, from the perspective of clinical dentistry, the pathway and histomorphometric characteristics of this nerve remain to be documented in detail. The purpose of the present study was to morphologically elucidate the pathway of the lingual nerve to clarify its significance in a clinical setting. A histomorphometric analysis was also performed in consideration of nerve grafting. The vertical distance between the occlusal plane and the superior margin of the lingual nerve showed a gradual decrease from the premolar toward the distal molar area. This suggests that the risk of injuring the lingual nerve increases gradually toward the distal area. The average total fascicular area of the lingual nerve was 1.90 mm2, which was larger than that of the sural nerve. It is the first-choice donor nerve for grafting. Therefore, even though the total fascicular area of the donor nerve is a little smaller than that of the recipient nerve, nerve grafting should be successful.

Histological study of the developing pterygoid process of the fetal mouse sphenoid.

The pterygoid process undergoes ossification of both the cartilage and membrane. However, few studies have attempted to explore the sequential development of the pterygoid process. Using histological examination, we performed morphological observations of the pterygoid process and surrounding tissue. ICR mice at embryonic days 13.5-18.0 and postnatal day 0 were used for morphological observations of the pterygoid process. By embryonic day 14.5, a mesenchymal cell condensation forming the anlage of the future medial pterygoid process differentiated into osteoid-like tissue and cartilage. At embryonic days 15.5-16.5, cartilage cells were clearly evident in the medial pterygoid process. In the medial pterygoid process, a bone collar was evident and calcified bone tissue surrounded the cartilage. At this point, a mesenchymal cell condensation formed the anlage of the pterygoid hamulus. At embryonic days 17.0-18.0, the cartilages were located along the lower and posterior border of the medial pterygoid process. A metachromatically stained matrix first became detectable around cells located in the pterygoid hamulus. On the other hand, at embryonic day 13.5, a metachromatically stained matrix was already evident in the space between the flattened cells in the lateral pterygoid process. At embryonic day 17.0, a hypertrophic cell zone had clearly formed in the diaphysis. On the basis of our present investigation, the lateral pterygoid process can be classified as primary cartilage, whereas the medial pterygoid process can be classified as secondary cartilage. Furthermore, it was found that the pterygoid hamulus is formed latest in the medial pterygoid process.

Sympathetic and parasympathetic neurons are likely to be absent in the human vestibular and geniculate ganglia: an immunohistochemical study using elderly cadaveric specimens.

The vestibular and geniculate ganglia of the ear in experimental animals carry both of the tyrosine hydroxylase (TH)-positive sympathetic neurons and the neuronal nitric oxide synthase (nNOS)-positive parasympathetic neurons. With an aid of immunohistochemistry, we examined these ganglia as well as the horizontal part of the facial nerve using specimens from 10 formalin-fixed elderly cadavers. The submandibular ganglion from the same cadavers was used for the positive control for both markers. Although there was a nonspecific reaction in nuclei for the present antibody of nNOS, these ganglia were unlikely to contain either nNOS- or TH-positive neurons. However, we did not deny a possibility that the absence was a result of degeneration with aging. In contrast, the facial nerve horizontal part consistently contained both of TH-positive- and nNOS-positive fibers. These fibers might regulate blood supply to the facial nerve and the dysregulation leads to edema to elevate pressure on the nerve within its osseous canal.

Synovial tissue morphology of the cricoarytenoid joint in the elderly: a histological comparison with the cricothyroid joint.

We compared the age-related morphology of the cricothyroid (CT) joint with that of the cricoarytenoid (CA) joint using 18 specimens from elderly cadavers in terms of their elastic fiber contents as well as the cells composing the joint capsule and synovial tissues. In contrast to an almost flat-flat interface in the CT joint, the CA joint was similar to a saddle joint. The CA joint capsule was thin and contained few elastic fibers, and in contrast to the CT joint, external fibrous tissues were not exposed to the joint cavity, there being no injury to the CA joint capsule. The lateral and posterior aspects of the CA joint were covered by the lateral and posterior CA muscles, respectively, and the fascia of the latter muscle was sometimes thick with abundant elastic fibers. However, due to possible muscle degeneration, loose connective tissue was often interposed between the fascia and the capsule. The medial and anterior aspects of the CA joint faced loose tissue that was continuous with the laryngeal submucosal tissue. Therefore, in contrast to the CT joint, a definite supporting ligament was usually absent in the CA joint. Synovial folds were always seen in the CA joint, comprising a short triangular mass on the posterior side and long laminar folds on the anterior side. The synovial folds usually contained multiple capillaries and a few CD68-positive macrophages. High congruity of the CA joint surfaces as well as strong muscle support to the arytenoid cartilage appeared to provide the specific synovial morphology.

Regional Anatomical Observation of Morphology of Greater Palatine Canal and Surrounding Structures.

In maxillary molar region implant therapy, support is sometimes obtained from trabecular bone comprising the maxillary tuberosity, pterygoid process of the sphenoid bone, and pyramidal process of the palatine bone. Great care is necessary in such cases due to the presence of the greater palatine canal, which forms a passageway for the greater palatine artery, vein, and nerve. However, clinical anatomical reports envisioning embedding of pterygomaxillary implants in this trabecular bone region have been limited in number. In this study, the 3-D morphology of the greater palatine canal region, including the maxillary tuberosity region and points requiring particular care in pterygomaxillary implantation, were therefore investigated. Micro-CT was used to image 20 dentulous jaws (40 sides) harvested from the dry skulls of Japanese individuals with a mean age of 28.2 years at time of death. The skulls were obtained from the Jikei University School of Medicine cadaver repository. Three-dimensional reconstruction of the trabecular bone region, including the greater palatine canal, was performed using software for 3-D measurement of trabecular bone structure. Trabecular bone region morphometry was performed with the hamular notch-incisive papilla (HIP) plane as the reference plane. The results showed a truncated-cone structure with the greater palatine foramen as the base extending to the pterygopalatine fossa. This indicates the need for care with respect to proximity of the dental implant body to the greater palatine canal and the risk of perforation if it is embedded in the maxillary tuberosity region at an inclination of 60° toward the lingual side. Moreover, caution must be exercised to avoid possible damage to the medial wall of the maxillary sinus if the inclination of the embedded dental implant body is almost perpendicular to the HIP plane.

Macrophage density in pharyngeal and laryngeal muscles greatly exceeds that in other striated muscles: an immunohistochemical study using elderly human cadavers / 대한해부학회지

Macrophages play an important role in aging-related muscle atrophy (i.e., sarcopenia). We examined macrophage density in six striated muscles (cricopharyngeus muscle, posterior cricoarytenoideus muscle, genioglossus muscle, masseter muscle, infraspinatus muscle, and external anal sphincter). We examined 14 donated male cadavers and utilized CD68 immunohistochemistry to clarify macrophage density in muscles. The numbers of macrophages per striated muscle fiber in the larynx and pharynx (0.34 and 0.31) were 5–6 times greater than those in the tongue, shoulder, and anus (0.05–0.07) with high statistical significance. Thick muscle fibers over 80 µm in diameter were seen in the pharynx, larynx, and anal sphincter of two limited specimens. Conversely, in the other sites or specimens, muscle fibers were thinner than 50 µm. We did not find any multinuclear muscle cells suggestive of regeneration. At the beginning of the study, we suspected that mucosal macrophages might have invaded into the muscle layer of the larynx and pharynx, but we found no evidence of inflammation in the mucosa. Likewise, the internal anal sphincter (a smooth muscle layer near the mucosa) usually contained fewer macrophages than the external sphincter. The present result suggest that, in elderly men, thinning and death of striated muscle fibers occur more frequently in the larynx and pharynx than in other parts of the body.