Yuji Ishida

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.

Occlusal hypofunction induces atrophic changes in rat gingiva.

OBJECTIVE To clarify the influence of occlusal hypofunction on the integrity of gingival tissue and gingival extracellular matrix biosynthesis. MATERIALS AND METHODS Thirteen-week-old male Wistar rats were divided into two groups. To eliminate occlusal forces, all the right maxillary molars were extracted in the hypofunctional group. The control group was anesthetized but not subjected to surgery. The rats were killed at 2 and 4 weeks after the procedure, and the lower right second molars were prepared for histological analysis. To investigate the effect of occlusal hypofunction on collagen biosynthesis, the expression of connective tissue growth factor (CTGF) and lysyl oxidase (LOX) was determined by immunohistochemistry as well as histological examination by hematoxylin and eosin staining. RESULTS Disorientation of the collagen fibers, proliferation of the connective tissue fibroblasts, and enlargement of epithelial intercellular gaps were observed in gingival tissue of rat molars with experimental occlusal hypofunction. Immunohistochemically, the expression of CTGF and LOX was increased significantly (P < .05) in the hypofunctional group. CONCLUSION These results suggest that occlusal hypofunction can affect the structural integrity and the expression of CTGF and LOX in gingival tissue.

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.

Unilateral maxillary molar extraction influences AQP5 expression and distribution in the rat submandibular salivary gland

OBJECTIVE Mastication has been regarded as a crucial factor for maintaining the morphology and function of secretion in salivary glands. Although it is known that occlusion affects mastication, the detailed process for how occlusal changes affect the secretory function of salivary glands is still unknown. Aquaporin 5 (AQP5) is a membrane protein that forms water channels, and plays an important role in water transport. In this study, we investigated the structural changes and alterations in the expression and distribution of AQP5 in the rat submandibular salivary gland (SMG) under occlusal hypofunction after unilateral molar extraction. METHODS Seven-week-old male Wistar rats (n = 36) were used in the study. In the experimental group, all of the right maxillary molars were extracted. Rats with no molar extraction were used as the control group. The rats were euthanized at 7, 14 or 28 days after the procedure, and the right SMGs were isolated and subjected to histological analyses. The expression and distribution of AQP5 were detected by immunohistochemistry. RESULTS Morphological analyses revealed hypertrophic changes in the acinar cells in the experimental group. Immunohistochemical staining of AQP5 was detected in the apical membrane (APM) and intercellular secretory canaliculi of acinar cells in both groups. On the other hand, the AQP5 expression in the APM and intercellular secretory canaliculi of acinar cells was less prominent in the experimental group than in the control group. CONCLUSIONS The present findings suggest that unilateral molar extraction has significant influences on the function of water transport in the rat SMG.

Effect of functional shift of the mandible on lubrication of the temporomandibular joint

Lubrication of synovial joints reduces the coefficient of friction of the articular cartilage surface. To investigate the effect of malocclusion on the lubrication of the temporomandibular joint (TMJ), we evaluated lubricin expression in the rat TMJ immunohistochemically, under conditions of functional lateral shift of the mandible, during period of growth. Thirty 5-week-old male Wistar rats were divided into experimental, recovery, and control groups. Each rt in the experimental and recovery groups was fitted with an acrylic-plate guiding appliance. The rats in the experimental and control groups were killed at 14 and 28 days after the appliance was attached. Each rat in the recovery group was detached from the appliance at 14 days, and was killed 14 days after the appliance was removed. In the experimental group, the expression of lubricin staining in TMJ cartilage was significantly decreased during the experimental period. In the recovery group, the expression of lubricin staining in TMJ cartilage was significantly greater than in the experimental group, and there was no significant difference at 28 days between the control and recovery groups. Analysis of these data suggests that a functional lateral shift of the mandible during the growth period influences lubrication of the TMJ.

Quantitative analysis of the relationship between maxillary incisors and the incisive canal by cone-beam computed tomography in an adult Japanese population

BackgroundIn setting goals for orthodontic treatment, determining the morphologies of the alveolar bone and maxillary incisor root is important for avoiding root resorption, dehiscence, and fenestration. This study aimed to analyze the configurational relationships among maxillary incisors, the alveolar border, and the incisive canal by cone-beam computed tomography (CBCT).MethodsCone-beam CT images of 93 orthodontic patients were evaluated for length of the incisive canal (L); angles between the palatal plane and the maxillary alveolar border (θ1), the incisive canal (θ2), and maxillary incisor (θ3); distance from the left maxillary incisor to the incisive canal (D); and cross-sectional areas of the incisive canal (CSAs) at three vertical levels. Comparison of variables between male and female patients was performed with the two-sample t test. Correlations between parameters were examined by Pearson’s correlation analysis and Bonferroni correction for multiple comparisons.ResultsMale patients exhibited significantly greater values of L than female patients. There were significant positive correlations between θ1 and θ2, θ2 and θ3, and θ3 and θ1. While the value of D was the lowest at the oral opening, that of the cross-sectional area of the incisive canal (CSA) was the greatest at the incisal root apex.ConclusionsThis study demonstrated that the incisive canal had large inter-individual variability, and the proximity between the incisive canal and the incisal root could not be precisely predicted by the conventional cephalograms. Therefore, pre-treatment CBCT examination should be recommended when a large amount of maxillary anterior retraction and/or intrusion is planned in orthodontic diagnosis.

A new approach to transfect NF-|[kappa]|B decoy oligodeoxynucleotides into the periodontal tissue using the ultrasound-microbubble method

The objective of this study is to investigate the effect of the ultrasound-microbubble technique in nuclear factor kappa B (NF-κB) decoy oligodeoxynucleotide (ODN) transfection in the gingival tissue in mice. The 6-FAM-labeled scrambled decoy ODN with microbubbles was applied to the periodontal tissue in 8-week-old male C57BL/6J mice by ultrasound radiation at low (LUM-Sc) and high (HUM-Sc) intensities to optimize the transfection condition of the ultrasound-microbubble method. Histological inspections were performed two hours after transfection to compare the expression with that in the sham-operated group without ultrasound radiation (A-Sc). Then, an NF-κB decoy was transfected into the periodontal tissue using the high-intensity ultrasound-microbubble (HUM-NF) technique to examine the anti-inflammatory effects of the decoy ODN. Western blot analysis was performed to investigate the expression of interleukin(IL)-1β, IL-6 and intercellular adhesion molecule-1 (ICAM-1) in the gingival tissues in the HUM-Sc, the HUM-NF and control groups. The fluorescence microscopy results showed that the fluorescent intensity in the periodontal tissues in the LUM-Sc and HUM-Sc groups was significantly higher than that in the A-Sc and the control groups. The fluorescent intensity in the HUM-Sc group, especially in the gingival connective tissue, was the highest of all groups. Western blot analysis indicated that the protein expression levels of IL-1β, IL-6 and ICAM-1 in the HUM-NF group were significantly lower than those in the HUM-Sc and the control groups. These findings suggest that the high-intensity ultrasound-microbubble technique is an effective tool for decoy transfection into the periodontal tissue.

Degeneration of fungiform and circumvallate papillae following molar extraction in rats

Abstract Objective. Proper occlusion facilitates food intake and gustatory function is indispensable for the enjoyment of food. Although an interaction between dentoalveolar and gustatory afferent neurons has been suggested by previous studies, the relationship between occlusion and gustation remains unclear. This study investigated the effect of upper molar extraction which diminished occlusal support on peripheral gustatory receptors in rats. Materials and methods. Thirty-six 7-week-old male Wistar rats were randomly assigned to either the experimental or the control group. All maxillary molars were extracted from rats in the experimental group under anesthesia, while a sham operation was conducted in the control group. The rats were euthanized 7, 14 or 28 days after the procedure. The morphology of the circumvallate papillae and taste buds using immunohistochemical methods and the fungiform papillae were visualized with 1% methylene blue. Results. Defects in the gustatory epithelium were observed after maxillary molar extraction. Rats in the experimental group had significantly fewer fungiform papillae, narrower circumvallate papillae, shallower trench depth, smaller trench area, smaller taste bud area, lower ratios of taste bud area to trench area and fewer taste buds than those in the control group. Conclusions. The findings indicate that molar extraction would affect peripheral gustatory receptors. This is the first study to characterize changes in rat fungiform and circumvallate papillae after maxillary molar extraction. This study suggests a possible synergic relationship between dentoalveolar perception and gustatory function, which has clinical implications that occlusion is closely correlated with gustatory perception.

Low-intensity pulsed ultrasound reduces periodontal atrophy in occlusal hypofunctional teeth

OBJECTIVE To clarify whether low-intensity pulsed ultrasound (LIPUS) exposure has recovery effects on the hypofunctional periodontal ligament (PDL) and interradicular alveolar bone (IRAB). MATERIALS AND METHODS Twelve-week-old male Sprague-Dawley rats were divided into three groups (n = 5 each): a normal occlusion (C) group, an occlusal hypofunction (H) group, and an occlusal hypofunction group subjected to LIPUS (HL) treatment. Hypofunctional occlusion of the maxillary first molar (M1) of the H and HL groups was induced by the bite-raising technique. Only the HL group was irradiated with LIPUS for 5 days. The IRAB and PDL of M1 were examined by microcomputed tomography (micro-CT) analysis. To quantify mRNA expression of cytokines involved in PDL proliferation and development, real-time reverse transcription quantitative PCR (qRT-PCR) was performed for twist family bHLH transcription factor 1 (Twist1), periostin, and connective tissue growth factor (CTGF) in the PDL samples. RESULTS Micro-CT analysis showed that the PDL volume was decreased in the H group compared with that of the C and HL groups. Both bone volume per tissue volume (BV/TV) of IRAB was decreased in the H group compared with that in the C group. LIPUS exposure restored BV/TV in the IRAB of the HL group. qRT-PCR analysis showed that Twist1, periostin, and CTGF mRNA levels were decreased in the H group and increased in the HL group. CONCLUSION LIPUS exposure reduced the atrophic changes of alveolar bone by inducing the upregulation of periostin and CTGF expression to promote PDL healing after induction of occlusal hypofunction.

The chemokine receptor type 4 antagonist, AMD3100, interrupts experimental tooth movement in rats

OBJECTIVE The aim of this study was to clarify the role of the stromal cell-derived factor-1 (SDF-1)/C-X-C chemokine receptor type 4 (CXCR4) axis in osteoclast accumulation, and the influence of orthodontic tooth movement (OTM) under mechanical force application to periodontal tissues, by administration of the CXCR4 antagonist AMD3100. DESIGN The upper right first molar (M1) of rats was moved mesially with a 10-g force titanium-nickel closed coil spring. Rats were treated with phosphate-buffered saline or AMD3100 (5mg/kg), which is a SDF-1 antagonist. After 0, 1, 3, and 7days, alveolar bones in all groups were examined at each time point by micro-computed tomography and histological analysis. RESULTS Tooth movement was decreased significantly in the AMD3100-treated group at 1, 3, and 7days after beginning OTM. The numbers of tartrate-resistant acid phosphatase-positive multinucleated cells in the periodontal ligament around the maxillary M1 were decreased significantly in the treated as compared to the control group on Days 1 and 3. CONCLUSION Administration of AMD3100 decreases OTM and osteoclast accumulation in rat molars under orthodontic force application. These findings suggest that the SDF-1/CXCR4 axis plays an important role in alveolar bone metabolism during OTM.