Autotransplantation of a Tooth Cryopreserved Over 11 Years Using a Programmed Freezer With a Magnetic Field: A Case Report.

This case report shows an autotransplantation of the lower right cryopreserved third molar into the extraction socket of the lower right first molar. Due to deep caries of the lower right first molar, the mesial root of this tooth was extracted. The patient asked to keep the distal root of the lower right first molar even if the root can survive only for a short period. So, a fixed partial denture supported by the lower right second premolar and the distal root of the lower right first molar was set. However, it was supposed that the distal root of the lower right first molar as an abutment tooth had a poor prognosis. Therefore, we also extracted the lower right third molar and cryopreserved to prepare autotransplantation if the lower right first molar has to be removed in the future. At first, the extracted third molar was frozen using a programmed freezer with a magnetic field named "Cells Alive System" (CAS) freezer, which was developed for tissue cryopreservation, and then, cryopreserved in the -150°C deep freezer. Eleven years later from the cryopreservation of the third molar, the lower right first molar showed root fracture. So, we extracted the lower right first molar and autotransplanted the cryopreserved third molar. Three years later, the autotransplanted tooth continued to be stable with healthy periodontium. The present case revealed that autotransplantation of a long-term cryopreserved tooth in a CAS freezer is a variable method to replace missing teeth.

Chewing ameliorates the effects of restraint stress on pERK-immunoreactive neurons in the rat insular cortex.

We investigated the effects of chewing on the anterior and posterior insular cortices during restraint stress using phosphorylated extracellular signal-regulated kinase (pERK) levels as a marker of neuronal responses. The stress only group demonstrated increased numbers of pERK-immunoreactive cells in both the anterior and posterior insular cortices compared to the control group (p < 0.01). In the stress with chewing group, the stress-induced increase of pERK-immunoreactive cell numbers was suppressed in both insular cortices and these differences were statistically significant compared to the stress-only group (p < 0.01). The suppressive effects of chewing were more prominent in the anterior insular cortex than in the posterior insular cortex. In general, the anterior insular cortex contributes to emotional processing, whereas the posterior insular cortex is associated with sensorimotor processes. Therefore, these results suggest that chewing ameliorates the emotional and sensorimotor responses to stress in the anterior and posterior insular cortices, respectively, with a greater effect on emotion-forming processes than on sensorimotor processes.

Mechanical and frictional properties of aesthetic orthodontic wires obtained by hard chrome carbide plating.

BACKGROUND/PURPOSE: Although aesthetic wire coating has been increasing in demand, it has problems that changes in mechanical properties and increase in frictional force. The aim of this study was to evaluate the coating of the wire, as characterized by aesthetics, in terms of low and constant friction and mechanical properties. MATERIALS AND METHODS: Hard chrome carbide-plated (HCCP) wires (HCCP group), commercially available polymer-coated wires (P group), rhodium-coated wires (R group), and uncoated wires (control group) were used. For all wire types, a stainless steel wire of dimensions 0.017 inch × 0.025 inch was used. They were evaluated by three-point bending, friction testing, surface observation, and colorimetric testing. RESULTS: The HCCP group was not significantly different from the control group in terms of flexural strength (σ) and flexural modulus (E) (σ: p = 0.90, E: p = 0.35). However, it was significantly inferior compared to the three other groups in terms of the maximum static and kinetic frictional forces under both dry and wet conditions (p < 0.05). In the surface observation, scratches were observed on the wire after the friction test. In the colorimetric test, no significant difference was observed between the HCCP group and the R group (p > 0.05). CONCLUSION: The mechanical properties of the HCCP wire were not significantly different compared to the control group. The frictional force of the HCCP wire was significantly lower than the other group. Therefore, the HCCP wire was suggested to increase the efficiency of tooth movement in clinics.

Adhesion of human periodontal ligament cells by three-dimensional culture to the sterilized root surface of extracted human teeth.

Residual periodontal ligament (PDL) and cement mass on the roots of extracted teeth are factors that considerably affect tooth transplantation. Therefore, when normal extracted teeth are used for autologous transplantation, it is necessary to regenerate the PDL of the root surface. Here we describe a method to examine human PDL cell adhesion on sterilized root surfaces. Sample teeth were extracted during orthodontic treatment. PDL cells were obtained from healthy periodontal tissue explants from teeth extracted for orthodontic reasons. We developed a method for adhering PDL cells to sterile root surfaces using three-dimensional culture for 3 weeks. We evaluated the adhesion of human PDL cells to the sterilized root surfaces biochemically and histologically. The adherent PDL cells presented new projections on the sterile root surfaces. Therefore, PDL cells can adhere to sterile root surfaces.

Irradiation effects of low temperature multi gas plasma jet on oral bacteria.

The purpose of this study is to evaluate the sterilization effects of a newly developed low temperature multi gas plasma jet on oral pathogenic microorganisms (Streptococcus mutans [S. mutans], Lactobacillus fermentum [L. fermentum], Aggregatibacter actinomycetemcomitans [A. actinomycetemcomitans]). Plasma gas which generated from O2, N2, Ar and 50% (O2+N2) was irradiated to the microbes. Effect of O2 plasma irradiation on S. mutans under scanning electron microscopy (SEM) was also observed. O2 plasma was directly applied to dental plaque on human extracted tooth. Then, the depth of enamel resorption area was noted by nanoscale hybrid microscope. O2 had the best sterilizing effect for all microbes. The potent bactericidal effect of plasma irradiation was also observed by SEM. Decalcification of enamel was noted significantly lower in plasma irradiated tooth surface compared to no plasma exposure group. These findings revealed that multi gas plasma jet has great potential to be used for dental treatment.

Simple Myofunctional Therapy Using Ready-made Mouthpiece Device before and after Orthodontic Treatment.

The present report describes myofunctional therapy using a ready-made training device, the T4A, in patients with permanent dentition and its effect on the prevention of relapse. The buccinator mechanism maintains the inner pressure of the tongue muscle equivalent to the outer pressure of the perioral soft tissues, such as the orbicular muscles, including the cephalopharyngeus and buccinator muscles. Training is performed so that patients learn to place their tongue and lips in the appropriate resting positions. The shape of the T4A and tongue guard supports the tongue from the bottom, allowing formation of the correct resting tongue position. However, the use of T4A for a long period of time may cause the teeth movement; therefore, caution is required. Use of the T4A is effective for the correction of oral habits, myofunctional therapy and for teaching the correct resting tongue position during the daytime and for the correction of oral habits, teaching correct resting tongue position during sleep.

Grinding patterns in migraine patients with sleep bruxism: a case-controlled study.

OBJECTIVES: Details on grinding patterns and types of contact during sleep bruxism in association with migraine headache have not yet been elucidated. This study compared the characteristics of sleep bruxism between patients with migraine and controls. METHODS: The study included 80 female patients who had been diagnosed with migraine and 52 women with no history of migraine. Grinding patterns were measured using the BruxChecker® (Scheu Dental, Iserlohn, Germany). RESULTS: There was a significant difference between the two groups in the distribution of grinding patterns at the laterotrusive side (p < 0.001). When the anterior teeth and premolar and molar regions in the two groups were compared, the proportion of the grinding area at all sites was significantly larger in the migraine group than in the control group (p < 0.001). DISCUSSION: The BruxChecker® showed that there was substantial grinding over a large area among migraine patients, particularly in the molar region.

Features of occlusal state in female Japanese patients with migraine: a case-controlled study.

OBJECTIVES: The prevalence rate of migraines is 8.4%; it is mostly diagnosed in women at 20s to 40s, and is known to cause major physical and mental disruption to daily life. This study was conducted on women aged between their 20s and 40s, in order to investigate the possible differences in the features of the occlusal state between a migraine and a non-migraine (control) group. METHODS: Age-matched female patients with migraine (n = 60) diagnosed by headache specialists and healthy controls (n = 57) were enrolled. Dental casts were used to evaluate some features. RESULTS: The maxillary and mandibular dentition casts from the migraine group showed significantly characteristic findings in their Angle's classification, overjet, and deviation in the anterior tooth midline, compared to the control group. DISCUSSION: The results relating occlusal state to both tension-related headaches and migraines, which have different pathogeneses, suggest the possibility of dental intervention to improve the symptoms of primary headaches.

Preoperative and postoperative examination of occlusal and maxillofacial changes after osteochondroma extirpation.

A patient came with left-side temporomandibular arthralgia, limited mandibular opening, frontal facial asymmetry, and a significant anterolateral open bite. Severe alterations in the occlusal and maxillofacial anatomy resulted from an osteochondroma associated with the mandibular condyle. We describe the changes associated with extirpation of the mandibular condylar osteochondroma and subsequent orthodontic treatment. These clinical changes resulted in improved facial symmetry and a satisfactory functional occlusion.

Prevention of root resorption in hypofunctional teeth by occlusal function recovery.

OBJECTIVE: To clarify whether occlusal hypofunction is one of the key determinants for root resorption during tooth movement and root resorption is prevented by its recovery. MATERIALS AND METHODS: The rats were randomly divided into one control and two experimental groups: hypofunctional and recovery groups. In the hypofunctional group, an anterior metal cap and bite plate were attached to the maxillary and mandibular incisors to simulate occlusal hypofunction. In the recovery group, the appliances were removed 7 weeks after their use, and the rats were allowed to bite for 4 weeks after removal. At the age of 16 weeks, the upper first molars were moved and after 0, 7, 14, and 21 days, the maxillae were resected. The resorption area was quantified morphohistologically and tartrate-resistant acid phosphatase (TRAP)-positive cells on the root surface were counted. We also examined the expressions of receptor activator of nuclear factor-κB ligand (RANKL), macrophage-colony stimulating factor (M-CSF), and interleukin (IL)-8 immunohistochemically. RESULTS: The amount of root resorption and the number of TRAP-positive cells were significantly greater in the hypofunctional group than in the control and recovery groups. Moreover, immunoreactivity for RANKL, M-CSF, and IL-8 was detected in the periodontal ligament and on the root surface in the hypofunctional group. CONCLUSION: Occlusal hypofunction is one of the critical factors for root resorption; however, root resorption may be prevented by recovery of occlusal function.

Effect of Melamine Sponge on Tooth Stain Removal.

OBJECTIVES: To investigate the stain removal ability of melamine sponge before aesthetic tooth whitening in extracted teeth. METHODS: Melamine sponge of thickness 40 mm was compressed and the destruction of the partition wall structure during the compression process was examined under a stereoscopic microscope. An extracted human tooth was cleaned by normal polishing or with melamine sponge for 90 s. To evaluate the stain level, the tooth surfaces were photographed under a stereoscopic microscope at 0, 30, 60 and 90 s. The residual stained region was traced in a high-magnification photograph, and the stain intensity was presented as a change, relative to the intensity before the experiment (0 s). RESULTS: Mechanical cleaning by toothbrushing produced polishing scratches on the tooth surface, whereas use of the melamine sponge resulted in only minimal scratches. As the compression level increased, the stain-removing effect tended to become stronger. CONCLUSION: Melamine sponge can remove stains from the tooth surface more effectively and less invasively compared to a conventional toothbrush. As no new scratches are made on the tooth surface when using a melamine sponge brush, the risk of re-staining is reduced. Cleaning using a melamine sponge brush can be easily and effectively performed at home and in a dental office.

Effects of mandibular retrusive deviation on prefrontal cortex activation: a functional near-infrared spectroscopy study.

The objective of this study was to evaluate occlusal condition by assessing brain activity in the prefrontal cortex, which is associated with emotion. Functional near-infrared spectroscopy (fNIRS) was used to detect changes in cerebral blood flow in the prefrontal cortex of 12 healthy volunteers. The malocclusion model was a custom-made splint that forced the mandible into retrusion. A splint with no modification was used as a control. The cortical activation during clenching was compared between the retrusive position condition and the control condition. A visual analog scale score for discomfort was also obtained during clenching and used to evaluate the interaction between fNIRS data and psychiatric changes. Activation of the prefrontal cortex was significantly greater during clenching in the mandibular retrusive condition than during clenching in the control condition. Furthermore, Spearman rank-correlation coefficient revealed a parallel relation between prefrontal cortex activation and visual analog scale score for discomfort. These results indicate that fNIRS can be used to objectively evaluate the occlusal condition by evaluating activity in the prefrontal cortex.

Chewing suppresses the stress-induced increase in the number of pERK-immunoreactive cells in the periaqueductal grey.

We investigated the effects of chewing under immobilization stress on the periaqueductal gray (PAG) matter using phosphorylated extracellular signal-regulated kinase (pERK) as a marker of responding cells. Immobilization stress increased pERK-immunoreactive cells in the PAG. Among four subdivisions of the PAG, the increase of immunoreactive cells was remarkable in the dorsolateral and ventrolateral subdivisions. However, increase of pERK-immunoreactive cells by the immobilization stress was not so evident in the dorsomedial and lateral subdivisions. The chewing under immobilization stress prevented the stress-induced increase of pERK-immunoreactive cells in the dorsolateral and ventrolateral subdivisions with statistical significances (p<0.05). Again, chewing effects on pERK-immunoreactive cells were not visible in the dorsomedial and lateral subdivisions. These results suggest that the chewing alleviates the PAG (dorsolateral and ventrolateral subdivisions) responses to stress.

A case of tooth autotransplantation after long-term cryopreservation using a programmed freezer with a magnetic field.

This case report describes the treatment of a skeletal Class III malocclusion with autotransplantation of a cryopreserved tooth. To gain an esthetic facial profile and good occlusion, extraction of bimaxillary premolars and surgical therapy were chosen. The patient had chronic apical periodontitis on the lower left first molar. Although she did not feel any pain in that region, the tooth was considered to have a poor prognosis. Therefore, we cryopreserved the extracted premolars to prepare for autotransplantation in the lower first molar area because the tooth would probably need to be removed in the future. The teeth were frozen by a programmed freezer with a magnetic field (CAS freezer) that was developed for tissue cryopreservation and were cryopreserved in -150°C deep freezer. After 1.5 years of presurgical orthodontic treatment, bilateral sagittal split ramus osteotomy was performed for mandible setback. Improvement of the facial profile and the occlusion were achieved in the retention phase. Six years after the initial visit, the patient had pain on the lower left first molar, and discharge of pus was observed, so we extracted the lower left first molar and autotransplanted the cryopreserved premolar. Three years later, healthy periodontium was observed at the autotransplanted tooth. This case report suggests that long-term cryopreservation of teeth by a CAS freezer is useful for later autotransplantation, and this can be a viable technique to replace missing teeth.

Ameliorating effects of Juzentaihoto on restraint stress and P. gingivalis-induced alveolar bone loss.

OBJECTIVE: Juzentaihoto (JTX) is a traditional Japanese medicine that consists of 10 herbs. The purpose of this study was to evaluate the efficacy of multi-herbal medicine JTX as a preventive and therapeutic drug for periodontal bone resorption and for reducing restraint stress. MATERIALS AND METHODS: Porphyromonas gingivalis ATCC 33277 was used for testing the antibacterial activity of JTX and a rat experimental periodontitis model. To evaluate the effect of JTX against P. gingivalis infection, we determined the differences in alveolar bone loss among experimental groups. The concentrations of adrenocorticotropic hormones were measured as stress markers, and atrophy of the thymus and spleen was assessed. RESULTS: JTX had antibacterial activity against P. gingivalis ATCC 33277. JTX treatment of mouse bone marrow cells at a concentration of 0.1 µg/ml significantly inhibited osteoclast formation. Administration of JTX to rats with P. gingivalis infection and restraint stress significantly reduced alveolar bone loss compared with the case with just the combination of P. gingivalis infection and restraint stress. In the restrained groups, stress markers were elevated, and the thymus and spleen were atrophied. The groups with administration of JTX showed not only inhibition of the decrease of weight but also normalization of corticosterone and cortisol values. CONCLUSION: JTX effectively inhibited restraint stress and osteoclastogenesis. It appears that the effects of JTX inhibit the destruction of periodontal tissue by suppressing stress. Our study demonstrated that JTX affects the correlation between restraint stress and periodontitis.

Effects of a low level laser on periodontal tissue in hypofunctional teeth.

Malocclusions, such as an open bite and high canines, are often encountered in orthodontic practice. Teeth without occlusal stimuli are known as hypofunctional teeth, and numerous atrophic changes have been reported in the periodontal tissue, including reductions in blood vessels in the periodontal ligament (PDL), heavy root resorption, and reduced bone mineral density (BMD) in the alveolar bone. Low Level Laser (LLL) has been shown to have a positive effect on bone formation and the vasculature. Although the recovery of hypofunctional teeth remains unclear, LLL is expected to have a positive influence on periodontal tissue in occlusal hypofunction. The aim of the present study was to elucidate the relationship between LLL and periodontal tissue in occlusal hypofunction. Twenty-four male rats aged 5 weeks were randomly divided into control and hypofunctional groups. An anterior metal cap and bite plate were attached to the maxillary and mandibular incisors in the hypofunctional group to simulate occlusal hypofunction in the molars. LLL irradiation was applied to the maxillary first molar through the gingival sulcus in half of the rats. Rats were divided into four groups; control, control+LLL, hypofunctional, and hypofunctional+LLL. Exposure to LLL irradiation was performed for 3 minutes every other day for 2 weeks. Animals were examined by Micro-CT at 5 and 7 weeks and were subsequently sacrificed. Heads were resected and examined histologically and immunohistologically. The hypofunctional group had obvious stricture of the PDL. However, no significant differences were observed in the PDL and alveolar bone between the hypofunctional+LLL and the control groups. In addition, the expression of basic fibroblast growth factor (bFGF) and vascular endothelial growth factor (VEGF)-positive cells were higher in the hypofunctional + LLL group than in the hypofunctional group. These results indicated that LLL enhanced the production of bFGF and VEGF in the periodontal tissue of hypofunctional teeth.

Cryopreservation of rat MSCs by use of a programmed freezer with magnetic field.

Mesenchymal stem cells (MSCs) can be used for the regeneration of various tissues and cryopreservation of MSCs is so important for regenerative medicine. The purpose of this study was to evaluate the influences of cryopreservation on MSCs by use of a programmed freezer with a magnetic field (CAS freezer). MSCs were isolated from bone marrow of rat femora. The cells were frozen by a CAS freezer with 10% dimethyl sulfoxide (Me2SO) and cryopreserved for 7 days at a temperature of -150 °C. Immediately after thawing, the number of survived cells was counted. The cell proliferation also examined after 48 h culture. Next, MSCs were frozen by two different freezers; CAS freezer and a conventional programmed freezer without magnetic field. Then, osteogenic and adipogenic differentiations of cryopreserved cells were examined. As a result, survival and proliferation rates of MSCs were significantly higher in CAS freezer than in the non-magnetic freezer. Alizarin positive reaction, large amount of calcium quantification, and greater alkaline phosphatase activity were shown in both the non-cryopreserved and CAS groups after osteogenic differentiation. Moreover, Oil Red O staining positive reaction and high amount of PPARγ and FABP4 mRNAs were shown in both the non-cryopreserved and CAS groups after adipogenic differentiation. From these findings, it is shown that a CAS freezer can maintain high survival and proliferation rates of MSCs and maintain both adipogenic and osteogenic differentiation abilities. It is thus concluded that CAS freezer is available for cryopreservation of MSCs, which can be applied to various tissue regeneration.

Open bite as a risk factor for orthodontic root resorption.

The purpose of the present study was to clarify the prevalence and degree of root resorption induced by orthodontic treatment in patients with and without open bite. One hundred and eleven patients treated with multibracket appliances were retrospectively selected from the patients and divided into non-open bite (NOB) and open bite (OB) groups. The severity of root resorption and the root shape were classified into five groups on periapical radiographs before and after treatment. Moreover, only in the OB group, all teeth were sub-divided into functional and hypofunctional ones that are occluding and non-occluding. As the results of multiple linear regression analysis of patient characteristics and clinical variables with the number of overall root resorption, the independent variables that were found to contribute significantly to root resorption were bite and abnormal root shape. The prevalences of root resorption evaluated in the number of patients were significantly higher in OB group than in NOB group, and those in the number of teeth were significantly higher in OB group than in NOB group, in particular anterior and premolar teeth. The prevalence of resorbed teeth with abnormal root shapes was also significantly higher in OB group than in NOB group. On the other hand, in OB group, the prevalences of root resorption and teeth with abnormal root shape were significantly greater in hypofunctional teeth than in normal functional teeth. There are more teeth with root resorption and abnormal root shape in open bite cases than in normal bite cases, and more teeth with abnormal root shapes and root resorption in hypofunctional teeth than in functional teeth.

Effects of vascular endothelial growth factor-C and -D on osteoclast differentiation and function in human peripheral blood mononuclear cells.

OBJECTIVE: The purpose of this study was to clarify the interaction of vascular endothelial growth factors (VEGFs)-C and -D with cell surface foetal liver kinase-1 (Flk-1) and fms-like tyrosine kinase-4 (Flt-4) receptors in the induction and activity of osteoclasts in cultured human peripheral blood mononuclear cells (PBMCs). DESIGN: PBMCs were cultured on chamber slides or on ivory discs for 2 or 3 weeks in the presence of macrophage-colony stimulating factor (M-CSF), VEGF-A, -C or -D, or placental growth factor (PlGF) with or without receptor activator of nuclear factor kappa-B ligand (RANKL). The number of osteoclasts in each group was counted and the area of ivory resorption was measured. In addition, osteoclast differentiation was further analysed under the same conditions, but with the addition of specific neutralizing antibodies against Flk-1 and Flt-4. RESULTS: RANKL was essential for the induction of osteoclasts in PBMCs. However, significant differences were found in the number of osteoclasts induced by VEGF-A, -C, -D or M-CSF with RANKL compared with control groups lacking or containing RANKL. Blocking of either Flk-1 or Flt-4 resulted in a reduction in the enhancement of osteoclast differentiation in PBMCs by VEGF-C or -D with RANKL. The osteoclasts induced by VEGF-A, -C, -D or M-CSF with RANKL formed significantly larger resorption lacunae than those formed by osteoclasts induced by RANKL alone. CONCLUSIONS: This study showed that VEGF-C and -D play a role in the induction of osteoclast differentiation through both Flk-1 and Flt-4 receptors and influence the area of the ivory resorption in PBMCs.