Biomedical subjects

Sadakazu Ejiri

Publications and source records attributed to Sadakazu Ejiri.

12 recordsLinked to original sources

PubMed2006

Histological and histomorphometrical changes in rat alveolar bone following antagonistic tooth extraction and/or ovariectomy.

OBJECTIVE: To examine changes appearing in the alveolar bone following the removal of the mechanical stress of occlusal loading, as well as the added influence of estrogen deficiency on such changes. DESIGN: The right mandibular molars of female rats were extracted. After 8 weeks, 12 animals were ovariectomized (OVX), and the other 12 were subjected to sham surgery (sham). Four weeks after surgery, all rats were sacrificed. The left-half and right-half maxillas of the sham group (the sham-occluded side and the sham-extruded side, respectively) and right-half maxilla of OVX group (the OVX-extruded side) were examined by histological observation and bone histomorphometry. RESULTS: The vertical height of alveolar bone in the sham-extruded and the OVX-extruded sides increased as compared with that of the sham-occluded side. In both extruded sides, active bone formation occurred on the surface of the alveolar bone facing the periodontal ligament, but the bone marrow was expanded and the bone volume had decreased in the internal area of the alveolar bone. In the OVX-extruded side, the bone marrow expanded more remarkably than that of the sham-extruded side, and the highest percentage of osteoclast surface was detected. CONCLUSIONS: Around the extruded teeth, there were regional differences in bone dynamics between the internal area of the alveolar bone and the bone surface facing the periodontal ligament, and estrogen deficiency seems to have caused further loss of bone volume in the interior of the alveolar bone supporting the extruded tooth.

Acid Phosphatase↗

PubMed2004

Osteocytic osteolysis observed in rats to which parathyroid hormone was continuously administered.

In order to prove osteocytic osteolysis in vivo, human parathyroid hormone (hPTH (1-34), 749 ng/h), or only solvent of the same volume, was continuously administered to 8-month-old rats by an infusion pump for 4 weeks, and then structural changes in osteocytes in the cortical bones of the tibiae were analyzed morphometrically, histologically, and histochemically. Based on contact microradiography (CMR) observations, the osteocyte lacunae in the PTH group tended to be enlarged, compared with those of the control, while the average lacuna area was 137.0 microm2 in the PTH group versus 93.9 microm2 in the control, suggesting evidence of osteocytic osteolysis. Acid phosphatase enzyme histochemical localization was observed in some osteocytes in the PTH group; therefore, lysosome systems may participate in the osteolytic mechanisms. On histological samples stained with hematoxylin-eosin or toluidine blue, the lacunae of the controls were surrounded by narrow areas of matrices both positive for hematoxylin and metachromatic for toluidine blue, while belt-like areas positive for hematoxylin were observed around the PTH-group lacunae. These findings suggested that, after osteocytic osteolysis, regenerated bone matrices may be added to the walls of osteocytes that possess enlarged lacunae.

Animals↗

PubMed2004

Calcitonin-induced change in serum calcium levels and its relationship to osteoclast morphology and number of calcitonin receptors.

It has been shown that, in live subjects, the ability of calcitonin (CT) to decrease serum calcium (Ca) levels can be lost in response to its continued or repeated administration. The present study investigated the relationship between such changes of in vivo serum Ca levels and the response of osteoclasts to CT administration, including the downregulation of their CT receptors (CTRs). Rats were either given a single injection of CT or repeated injections at either 6- or 24-h intervals, after which their serum Ca levels were evaluated. Their parietal bones were dissected, and the amount of 125I-labeled elcatonin (125I-eCT) binding to their osteoclasts measured using autoradiography. Ultrastructural changes in the osteoclasts were also examined. Twenty-four hours after a single CT administration, serum Ca levels had dropped, and there was an absence of ruffled borders on the osteoclasts. Less 125I-eCT binding to the osteoclast was found than in the control group. Forty-eight and 72 h after CT administration, serum Ca levels had almost returned to control levels, and the osteoclasts showed ruffled borders once again. The amount of 125I-eCT binding to the osteoclast also recovered to control levels. When these osteoclasts were then incubated in CT, their ruffled borders once again disappeared. In the 6-h interval multiple CT administration schedule subjects, upon inspection 72 h after their first administration (6 h following the final one), serum Ca levels were found to have almost returned to control levels with the presence of osteoclast ruffled borders. The amount of 125I-eCT binding to these osteoclasts was remarkably limited, and no disappearance of the ruffled borders occurred in response to additional CT incubation. In the 24-h interval multiple administration schedule subjects, upon inspection 72 h after their first CT administration (24 h following the final one), there was less 125I-eCT binding than in the single-dose subjects tested 24 h after their injection, and the ability of CT to lower their serum Ca levels was reduced. The ability of CT to lower serum Ca levels was therefore related to the response of osteoclasts to the CT (the disappearance of the ruffled borders), and this response was related to the amount of CTRs available for binding with CT on the osteoclast surface. Furthermore, the reduced effectiveness of CT in response to repeated CT administration was found to be related to the downregulation of the CTRs on the osteoclast surface.

Animals↗

PubMed2003

Tensile stress induces alpha-adaptin C production in mouse calvariae in an organ culture: possible involvement of endocytosis in mechanical stress-stimulated osteoblast differentiation.

We previously demonstrated that tensile stress (TS)-induced osteoblast differentiation eventually led to osteogenesis in an organ culture of mouse calvarial sutures. In the present study, we employed RNA-fingerprinting using an arbitrarily primed polymerase chain reaction (RAP-PCR) to identify alpha-adaptin C, a component of the endocytosis machinery AP2, as a TS-inducible gene. Protein production, as well as the gene expression of alpha-adaptin C, was induced by TS as early as 3 h following the initiation of loading. In situ hybridization and immunohistochemical analysis revealed that the induction of alpha-adaptin C mostly occurred in fibroblastic cells in the sutures, suggesting that it precedes TS-induced osteoblast differentiation. Consistent with this result, TS significantly increased the number of coated pits (CPs) and coated vesicles (CVs) in the undifferentiated fibroblastic cells but not in the osteoblastic cells around calvarial bones. Further, TS-induced osteoblast differentiation was suppressed when endocytosis was inhibited by potassium depletion. These results, taken together, suggest that TS accelerates osteoblast differentiation and osteogenesis, possibly through the induction of the alpha-adaptin C expression and consequent activation of receptor-mediated endocytosis.

Adaptor Protein Complex 2↗

PubMed2003

Bone structure and mineralization demonstrated using synchrotron radiation computed tomography (SR-CT) in animal models: preliminary findings.

We investigated the ability of synchrotron radiation computed tomography (SR-CT) to demonstrate trabecular microstructure, detail of trabecular surfaces, and mineralization of bones. Eight rat vertebrae, six rat tibiae, and eight minipig vertebrae were scanned using SR-CT at the synchrotron radiation facility Super Photon ring-8GeV (SPring-8). Images obtained using conventional micro-CT, scanning electron microscopy (SEM), and contact microradiography (CMR) were compared with the SR-CT images. SR-CT showed high image quality without visible partial volume effect. Three-dimensional SR-CT revealed shallow concavities in the bone surface, which were considered to correspond to osteoclastic resorption areas, as well as the connectivity, anisotropy, and shape (rod- or platelike) of trabeculae. Two-dimensional SR-CT showed different density along the surface of the trabecular bone, indicating the degree of bone mineralization. In conclusion, SR-CT seems to be a useful tool for delineating trabecular surfaces, evaluating bone mineralization, and revealing precise trabecular structure.

Animals↗

PubMed2003

Osteoclast differentiation in ectopic bone formation induced by recombinant human bone morphogenetic protein 2 (rhBMP-2).

Osteoclast differentiation in the process of ectopic bone formation induced by recombinant human bone morphogenetic protein 2 (rhBMP-2) was examined to clarify the relationship between osteoclast development and rhBMP-2-induced bone formation. A combination of rhBMP-2 with a porous microsphere (PMS) and blood clot was implanted subcutaneously on the bilateral chest muscles of rats. Tartrate-resistant acid phosphatase (TRAPase) activity, cathepsin K (cath K), and calcitonin receptor (CTR), as markers of osteoclasts and their precursors, were examined using enzyme and immunohistochemical analysis up to 7 days after implantation. Mononuclear cells positive for TRAPase, cath K, and CTR first appeared on day 3 in connective tissue surrounding the PMS after implantation of rhBMP-2. Simultaneously, alkaline phosphatase activity became detectable in mesenchymal cells in the connective tissue. Electron microscopy demonstrated some mononuclear cells with abundant mitochondria and poorly developed rough endoplasmic reticulum in the proximity of mesenchymal cells. However, there was no evidence of cartilage or bone matrix formation on day 3. Osteoclasts in various stages of development, classified by the pattern of immunoreactivity for cath K, were observed by day 7. The polarized intracellular distribution of cath K was found only in osteoclasts attached to bone matrix. In conclusion, we have demonstrated for the first time the appearance of osteoclast precursors before bone matrix formation induced by rhBMP-2, suggesting that bone matrix is not a prerequisite for osteoclast precursor recruitment. Furthermore, we suggest that differentiation into polarized functional osteoclasts is accomplished when the osteoclasts attach to the bone matrix.

Animals↗

PubMed2003

Bisphosphonate incadronate inhibits maturation of ectopic bone induced by recombinant human bone morphogenetic protein 2.

To determine the effects of a bisphosphonate on the quality of bone morphogenetic protein-(BMP-) induced bone, incadronate was administered to rats in which subcutaneous ectopic bones were induced by recombinant human BMP-2. Incadronate (1 microg/kg/day) was administered to rats carrying the BMP-induced bones three times per week, from the 3rd to 7th week after BMP implantation (incadronate group). Aliquots of phosphate-buffered saline were administered in the same protocol without incadronate to the control group. During the 3rd, 4th, 7th, or 10th week, the BMP-induced bones were removed and observed by contact microradiography (CMR), H&E staining, enzyme histochemistry for tartrate-resistant acid phosphatase (TRAP), and immunohistochemistry for cathepsin K. By 3 weeks when administration of incadronate began, woven bones formed in the periphery of the BMP pellets and osteoclasts were attached to these bones. At 4 weeks, in both the incadronate and control groups, bone formation advanced inward. However, in the incadronate group, poorly calcified areas, corresponding to the remaining BMP pellets, were found in the middle areas of bone formation, whereas osteoclasts decreased when compared with those of the control group. During the 10 weeks, bone marrow was formed and the characteristics of lamellar bones, in which the lacunae of small osteocytes were regularly arranged, were noted in the control group. In contrast, the poorly calcified areas were still present up to 10 weeks in incadronate group in which the osteoclasts were also scarcer than in the control group. These findings suggested that osteoclast-mediated bone resorption was inhibited by incadronate administration, and that immature bones including the BMP pellets remained for a long time, indicating that the process of bone maturation was blocked. The possibility of using incadronate for the purpose of inhibiting osteoclastic bone resorption was confirmed, but further study is needed before clinical application.

Acid Phosphatase↗

PubMed2003

Long-term changes in trabecular structure of aged rat alveolar bone after ovariectomy.

OBJECTIVE: Trabecular structural changes in the jaw after long-term estrogen deficiency are not well understood. Therefore, we sought to observe the changes in rat alveolar bone for 1 year. METHODS: Six-month-old female rats were ovariectomized (OVX) or underwent a sham operation. After 1 year, bone histomorphometry and a node-strut analysis were performed on the interradicular septum of the mandibular first molar by using micro computed tomography and confocal laser scanning microscopy. Statistical analysis was carried out by using analysis of variance. RESULTS: The alveolar trabeculae of rats in the sham group had network structures, whereas the trabeculae of rats in the OVX group became fragmented. The trabecular bone volume, number, and thickness in the OVX group were significantly lower than those found in the sham group, and the trabecular separation was 4-fold higher in the OVX group than in the sham group. Bone resorptive and formative activity appeared to be moderately higher in the OVX group than in the sham group, but only the difference in bone formation was of statistical significance. CONCLUSION: By 1 year after ovariectomy, bone loss and trabecular fragmentation had occurred in the rat mandibular alveolar bone.

Aging↗

PubMed2003

[Effects of ovariectomy on trabecular structures of rat alveolar bone].

To clarify the relationship between estrogen deficiency and tooth loss, we analyzed the trabecular structural changes of mandibular alveolar bone in ovariectomized rats. Two months after ovariectomy, the bone resorptive activity remarkably accelerated and caused high bone turnover. On the other hand, one year after ovariectomy, bone loss and trabecular fragmentation occurred in the alveolar bone, although the accelerated degree of bone resorptive activity was moderate. If elderly women could experience similar changes on their alveolar bone, we must pay more attention to the bone resorption presently after menopause and the trabecular fragility long after menopause.

English Abstract↗

PubMed2002

Odontoblast processes in human dentin revealed by fluorescence labeling and transmission electron microscopy.

In the present undertaking, the distribution of odontoblast processes in human dentin was determined through the DiI carbocyanine dye fluorescent staining of the cell membrane, while F-actin was identified by rhodamine-phalloidin. Confocal laser scanning microscopy revealed intense labeling for both agents in inner dentin, while transmission electron microscopy (TEM) identified dentinal tubules including odontoblast processes in this area, each process being surrounded by a cell membrane and containing an abundance of filamentous structures. Electron-dense "lamina limitans" lined the dentinal tubules. Individual cell processes became narrower toward the middle area, and their overall numbers decreased as well under TEM. Labeling for F-actin was absent in both middle and outer dentin, while faint labeling for DiI was visible along the dentinal tubules as far as the dentino-enamel junction (DEJ), where it was also recognized within the tubules themselves. Under TEM, the dentinal tubules lined with electron-dense structures were, in fact, empty in the middle and outer dentin. Immediately below the DEJ, however, the tubules manifested dense concentrations of fine granular material. Our study, therefore, appears to suggest that odontoblast processes do not extend beyond the inner dentin of fully erupted human premolars.

Actins↗

PubMedDate not supplied

Bone regeneration by recombinant human bone morphogenetic protein-2 around immediate implants: a pilot study in rats.

PURPOSE: Difficulties relating to bone regeneration that complicate immediate implant placement include buccal and/or lingual fenestrations, primary anchorage of the implants, and the need for protection from functional loading during the osseointegration period. The objective of this pilot study was to evaluate bone regeneration by recombinant human bone morphogenetic protein-2 (rhBMP-2) around immediate implants placed in maxillary sockets in rats. MATERIALS AND METHODS: A total of 16 cylindric 0.8 x 1.8-mm commercially pure, solid titanium Implants were placed immediately after gentle extraction of the maxillary first molar teeth of 8 male Wistar rats. The sockets were randomly divided into 3 groups: group 1 (n = 6) received rhBMP-2 with polylactic acid/polyglycolic acid copolymer-coated gelatin sponge carrier; group 2 (n = 5) received only the carrier; and group 3 (n = 5) received no grafting materials following placement The rats were euthanized at 90 days postsurgery for microscopic analysis. RESULTS: In group 1, the implant body remained submerged completely, including the coronal part, which was fully covered by a significant amount (30% of total height) of regenerated cortical bone, even though the implant could easily be pulled out by a tweezer at the time of placement. Close approximation between the implant surface and regenerated bone could also be detected, indicating good bone-to-implant contact. In contrast, only peri-implant bone regeneration occurred in group 2, and an approximate 0.3-mm coronal part of the implant remained exposed. When no grafting materials were used (group 3), almost one third of the total length of the implant was exfoliated out of the socket when no grafting materials were used. DISCUSSION AND CONCLUSIONS: Based on previous study and data from 16 sockets of the present study, it could be concluded that rhBMP-2 facilitated the regeneration of bone around immediate implants. In particular, the bone covering the coronal part could have been regenerated shortly after surgery, which helped to maintain the implant body inside the socket during the integration period in rats.

Animals↗

PubMedDate not supplied

Peri-implant stress analysis in simulation models with or without trabecular bone structure.

Most 3-dimensional (3D) finite element analyses (FEAs) simplify the cancellous bone to a block and completely ignore its trabecular structure. Thus, a 3D FEA was performed to compare the peri-implant stress distribution of a model in which the trabecular structure was accurately simulated (precise model) with that of a model with a homogenous cancellous bone component (simplified model). In contrast to the simplified model, the distribution patterns and higher stresses in the precise model may explain the overall bone resorption at the implant-bone interface in load-related implant failures. Further studies using data from the jawbone and a more detailed implant simulation are planned.

Animals↗