Jun Hosomichi

Effect of sympathetic nervous activity on alveolar bone loss induced by occlusal hypofunction in rats

OBJECTIVE To elucidate the effect of sympathetic nervous activity on alveolar bone loss induced by occlusal hypofunction in rat molars. DESIGN Occlusal hypofunction in the molar area was produced by attaching appliances to rat maxillary and mandibular incisors. In addition, a non-selective β-adrenergic receptor antagonist, propranolol, was administered orally to rats in drinking water to pharmacologically suppress sympathetic nervous activity. After 1 week, alveolar bones in all groups were examined by micro-CT, histomorphometry and histology to determine their trabecular bone phenotypes and histological changes. RESULTS The marrow spaces of the interradicular alveolar bone of rat mandibular first molars (M1) increased in the occlusal hypofunction group (Group H) but not in the control group (Group C), whilst these decreased in rats in the occlusal hypofunction group that were administered propranolol (Group HB). Bone volume/tissue volume (BV/TV), trabecular thickness (Tb.Th) and trabecular number (Tb.N) for interradicular alveolar bone in M1 in Group H were significantly lower than those in Group C, whereas those in Group HB remained as high as those in Group C. The number of TRAP-positive cells in Group H increased compared to that in Group C, whereas it significantly decreased in Group HB. CONCLUSIONS These results suggest that sympathetic nervous activity may influence the alveolar bone loss induced by occlusal hypofunction.

Soft diet causes greater alveolar osteopenia in the mandible than in the maxilla

OBJECTIVE To investigate changes in the bony microstructure of the upper and lower alveolar bone during masticatory loading induced by soft diet feeding in growing rats. DESIGN Three-week-old male Wistar rats were randomly divided into two groups. Rats were fed with either pellets [control group (n=6)] or a soft diet [experimental group (n=6)] for nine weeks. 3D-microstructure of the alveolar bone of the first molar region (M1) was examined by micro-CT analysis. RESULTS Micro-CT images showed increased marrow spaces of the inter-radicular alveolar bone around the rat mandibular M1 in the experimental group compared with that in the control group. The bone volume/tissue volume ratio, trabecular thickness, trabecular number, mean intercept length, trabecular width and trabecular star volume for the mandibular M1 inter-radicular alveolar bone were lower in the experimental group than in the control group. Marrow space star volume was increased in the experimental group compared with the control group. CONCLUSIONS These results suggest that alveolar osteopenia is more extensive in the mandible than the maxilla in rats that experience low masticatory loading during growth.

Occlusal hypofunction causes periodontal atrophy and VEGF/VEGFR inhibition in tooth movement

OBJECTIVE To examine changes in microvasculature and the expression of vascular endothelial growth factor A (VEGF-A) and VEGF receptor 2 (VEGFR-2) in rat hypofunctional periodontal ligament (PDL) during experimental tooth movement. MATERIALS AND METHODS Twelve-week-old male Sprague-Dawley rats were divided into normal occlusion and occlusal hypofunction groups. After a 2-week bite-raising period, rat first molar was moved mesially using a 10-gf titanium-nickel alloy closed coil spring in both groups. On days 0, 1, 2, 3, and 7 after tooth movement, histologic changes were examined by micro-computed tomography and immunohistochemistry using CD31, VEGF-A, VEGFR-2, and the terminal deoxynucleotidyl transferase mediated dUTP nick end labeling (TUNEL) method. RESULTS Hypofunctional molars inclined more than normal molars and did not move notably after day 1 of tooth movement. Blood vessels increased on the tension side of the PDL in normal teeth. Immunoreactivities for VEGF-A and VEGFR-2 in normal teeth were greater than those in hypofunctional teeth during tooth movement. Compressive force rapidly caused apoptosis of the PDL and vascular endothelial cells in hypofunctional teeth, but not in normal teeth. CONCLUSIONS Occlusal hypofunction induces vascular constriction through a decrease in the expression of VEGF-A and VEGFR-2, and apoptosis of the PDL and vascular cells occurs during tooth movement.

Intermittent posterior displacement of the rat mandible in the growth period affects the condylar cancellous bone

OBJECTIVE To examine whether intermittent posterior condylar displacement causes changes in cancellous bone in the mandibular condyle during the growth period. MATERIALS AND METHODS Sixteen 5-week-old male Wistar rats were divided into experimental and control groups. In the experimental group, an appliance was attached to the maxillary incisors to induce posterior displacement of the condyles in the occluded condition. Untreated rats served as the control group. Animals were sacrificed at 14 days, and the condyles were removed to analyze the three-dimensional cancellous bone structure by microcomputed tomography (micro-CT). Serial sagittal paraffin sections of the condyles were used for hematoxylin and eosin staining to investigate histomorphological changes and for tartrate-resistant acid phosphatase (TRAP) staining to identify osteoclastic cells. RESULTS Micro-CT analysis showed that in the experimental group, the bone volume fraction and the degree of anisotropy were significantly decreased compared with those in the control group in the anterior region of the condyle. Moreover, the number of TRAP-positive cells was significantly greater in the same region in the experimental group than in the control group. CONCLUSION Intermittent posterior displacement of the mandible can cause region-specific changes in the profile and microarchitecture of the condylar cancellous bone.

Micro-computed tomography analysis of changes in the periodontal ligament and alveolar bone proper induced by occlusal hypofunction of rat molars

Objective To three-dimensionally elucidate the effects of occlusal hypofunction on the periodontal ligament and alveolar bone proper of rat molars by micro-computed tomography (micro-CT). Methods Occlusal function in the molar area was restricted by attaching an anterior bite plate on the maxillary incisors and a metal cap on the mandibular incisors of 5-week-old male Wistar rats for 1 week. The periodontal ligament space and alveolar bone proper around roots of the mandibular first molar were assessed by histology and micro-CT. Results The periodontal ligament space was narrower and the alveolar bone proper was sparser and less continuous in the hypofunction group than in the control group. Further, both the volume of the periodontal ligament and the volumetric ratio of the alveolar bone proper to the total tissue in the region of interest were significantly lower in the hypofunction group (p < 0.05). Conclusions Occlusal hypofunction induces atrophic changes in the periodontal ligament and alveolar bone proper of rat molars.

Intermittent hypoxia induces disturbances in craniofacial growth and defects in craniofacial morphology.

OBJECTIVES To investigate intermittent hypoxia (IH) induced changes in craniofacial morphology and bone mineral density (BMD) in the mandible of growing rats. DESIGN Seven-week-old male Sprague-Dawley rats were exposed to IH for 4 days or 3 weeks. Sham-operated rats simultaneously breathed room air. Lateral and transverse cephalometric radiographs of the craniofacial region were obtained, and the linear distances between cephalometric landmarks were statistically analyzed. BMD and bone microstructure of the mandible were evaluated using micro-computed tomography (micro-CT). RESULTS Cephalometric analyses demonstrated that exposure to IH only in the two groups for 3 weeks decreased the size of the mandibular and viscerocranial bones, but not that of the neurocranial bones, in early adolescent rats. These findings are consistent with upper airway narrowing and obstructive sleep apnea (OSA). Micro-CT showed that IH increased the BMD in the cancellous bone of the mandibular condyle and the inter-radicular alveolar bone in the mandibular first molar (M1) region. CONCLUSIONS This study is the first to identify growth retardation of the craniofacial bones in an animal model of sleep apnea. Notably, 3 weeks of IH can induce multiple changes in the bones around the upper airway in pubertal rats, which can enhance upper airway narrowing and the development of OSA. The reproducibility of these results supports the validity and usefulness of this model. These findings also emphasize the critical importance of morphometric evaluation of patients with OSA.

Influence of Occlusal Stimuli on the Microvasculature in Rat Dental Pulp

OBJECTIVE To examine the influence of occlusal stimuli on the vasculature in the dental pulp, using an occlusal hypofunction model. MATERIALS AND METHODS Twenty 7-week-old male Sprague-Dawley rats were divided into two groups. To produce occlusal hypofunction, the appliances were attached to the maxillary and mandibular incisors. Untreated rats served as controls. Serial horizontal paraffin sections of the mandibular first molar were processed by conventional methods. To evaluate the microvasculature in the dental pulp, sections of each specimen were stained with hematoxylin-eosin. RESULTS In the experimental group, the arterioles in the tooth pulp tissue ran convergently, and their inside diameter was significantly smaller than that of the control group. CONCLUSION This study suggests that occlusal stimuli influence the periodontal ligament throughout the microvasculature of the dental pulp.

Influence of occlusal hypofunction on the elastic property and bone formation of rat alveolar bone

Abstract The purpose of this study was to investigate the influence of occlusal hypofunction on the maintenance of the elastic property and alveolar bone formation using an occlusal hypofunction model. Analyses of the elastic property and bone formation of the alveolar bone were carried out by the compression test and the bone histomorphometry in the two directions (the tooth axis direction and the bucco-lingual direction). The compression test was done with the texturometer to analyse the springiness of bone specimens of the mandibular first molar. Bone histomorphometric analyses were evaluated using double labeled frontal section. Results showed that occlusal hypofunction significantly reduced the springiness and suppressed the alveolar bone formation in the bucco-lingual direction. These results suggested that occlusal function plays an important role in maintenance of the elastic property and bone formation of the alveolar bone.

ALTERATION OF BFGF EXPRESSION WITH GROWTH AND AGE IN RAT MOLAR PERIODONTAL LIGAMENT

OBJECTIVE To test the hypothesis that immunohistochemical changes in expression of basic fibroblast growth factor in the periodontal ligament do not change with age. MATERIALS AND METHODS Thirty male Wistar-ST rats were divided into growing groups (5, 9, and 15 weeks of age) and aging groups (6, 12, and 18 months of age). Serial sagittal sections (5 microm thick) were cut parallel at the distobuccal roots of the maxillary first molar. Paraffin-embedded tissue sections were stained with hematoxylin and eosin and rabbit polyclonal antibodies to basic fibroblast growth factor. RESULTS The number and the area of basic fibroblast growth factor-immunoreactive cells in the periodontal ligament of the maxillary first molar decreased with age. The number of basic fibroblast growth factor-immunoreactive cells was much greater in the root furcation area, which experiences the greatest effect of occlusal force. Regardless of age, the production of basic fibroblast growth factor in the periodontal ligament may occur subject to functional demand. CONCLUSIONS The hypothesis is rejected. The expression of basic fibroblast growth factor in the periodontal ligament decreased with age.

Intermittent Hypoxia Influences Alveolar Bone Proper Microstructure via Hypoxia-Inducible Factor and VEGF Expression in Periodontal Ligaments of Growing Rats

Intermittent hypoxia (IH) recapitulates morphological changes in the maxillofacial bones in children with obstructive sleep apnea (OSA). Recently, we found that IH increased bone mineral density (BMD) in the inter-radicular alveolar bone (reflecting enhanced osteogenesis) in the mandibular first molar (M1) region in the growing rats, but the underlying mechanism remains unknown. In this study, we focused on the hypoxia-inducible factor (HIF) pathway to assess the effect of IH by testing the null hypothesis of no significant differences in the mRNA-expression levels of relevant factors associated with the HIF pathway, between control rats and growing rats with IH. To test the null hypothesis, we investigated how IH enhances mandibular osteogenesis in the alveolar bone proper with respect to HIF-1α and vascular endothelial growth factor (VEGF) in periodontal ligament (PDL) tissues. Seven-week-old male Sprague–Dawley rats were exposed to IH for 3 weeks. The microstructure and BMD in the alveolar bone proper of the distal root of the mandibular M1 were evaluated using micro-computed tomography (micro-CT). Expression of HIF-1α and VEGF mRNA in PDL tissues were measured, whereas osteogenesis was evaluated by measuring mRNA levels for alkaline phosphatase (ALP) and bone morphogenetic protein-2 (BMP-2). The null hypothesis was rejected: we found an increase in the expression of all of these markers after IH exposure. The results provided the first indication that IH enhanced osteogenesis of the mandibular M1 region in association with PDL angiogenesis during growth via HIF-1α in an animal model.