The Physiological Function of Deglutition and Swallowing in Patients With Maxillary Protrusion After Mandibular Advancement.

The objective of this study was to determine the tongue-palatal contact changes in patients with skeletal maxillary protrusion after sagittal split ramus osteotomy (SSRO) during swallowing. In this study, 15 patients with maxillary protrusion and 10 normal subjects participated. Before and 3 months after surgery, tongue-palatal contact patterns during swallowing of patients with maxillary protrusion as well as controls were evaluated by electropalatography. The electrode contact number in the alveolar, palatal, and velar parts was examined. The swallowing duration of each phase was also evaluated. In the lateral area of the velar part, incomplete electrode contact was shown at 0.3 seconds in patients with maxillary protrusion. The electrode contact number in the velar part at 0.3 seconds before tongue-palatal complete contact was significantly less in the preoperative patients compared with the controls ( P < 0.05). A small increase in the electrode contact number of the velar part was shown in the postoperative patients at 0.3 and 0.2 seconds before tongue-palatal complete contact ( P < 0.05). The pharyngeal phase duration was significantly larger in the patients with maxillary protrusion before SSRO compared with the controls ( P < 0.05). After SSRO, the pharyngeal phase duration was significantly shortened. It was shown that the tongue-palatal contact pattern during swallowing in patients with maxillary protrusion improved after orthognathic surgery, and the pharyngeal phase duration was also shortened. It is suggested that the changes in the mesiodistal mandibular position by orthognathic surgery can improve tongue posture and movement during swallowing.

Correction of Severe Skeletal Class II High Angle with Mandibular Retrusion and Gummy Smile by Double-Jaw Surgery.

This report describes the treatment of severe skeletal Class II malocclusion in a young woman with a gummy smile and pronounced lower anterior facial height. Overjet and overbite were +12.0 mm and -1.0 mm, respectively. Cephalometric analysis revealed inferior positioning of the maxilla and severe mandibular retrusion with clockwise rotation. Both the upper and lower anterior teeth showed labial inclination. Based on a diagnosis of a skeletal Class II high angle with mandibular retrusion and a gummy smile, double-jaw orthognathic surgeries for upper and lower premolar extraction were chosen to gain ideal occlusion and an improvement in the esthetic facial profile. Le Fort I osteotomy was performed to move the anterior and posterior teeth upward by 4.0 mm and achieve mandibular counterclockwise rotation. Short lingual sagittal split ramus osteotomy was performed to move the mandible forward by 3.0 mm. As a result, normal overjet and overbite were achieved together with a straight profile and a good smile. After surgery, electromyographic evaluation of anterior temporal muscle activity showed an improvement in the percentage overlapping coefficient value (a symmetric index of bilateral muscle activity) from 28.1% to 63.2% compared to at pre-treatment. The pattern of jaw movement also showed an improvement. These results suggest that orthognathic surgery in skeletal Class II cases can improve not only malocclusion and the skeletal relationship of the jaws, but also masticatory function and jaw movement.

Lansoprazole and zoledronate delays hard tissue healing of tooth extraction sockets in dexamethasone-treated mice.

Proton pump inhibitors (PPIs) are among the most commonly prescribed medicines for the management of acid-related gastrointestinal diseases. Osteonecrosis of the jaw (ONJ) is a serious adverse event that is associated with the use of antiresorptive and antiangiogenic agents. According to previous clinical reports, the use of PPIs contributes to the pathogenesis of severe ONJ that requires surgery. Here, we investigated the effects of lansoprazole (LP) or LP in combination with zoledronate (ZOL) on ONJ development in mice. C57BL/6J mice were administered ZOL (125 µg/kg intravenously, twice weekly) and/or LP (10 mg/kg intraperitoneally; 3 weeks of 3 consecutive days followed by 1 day off). One week after initiation of the study, the first molar was atraumatically extracted. Concurrently with ZOL administration, dexamethasone (Dex) was administered (5 mg/kg intraperitoneally, twice weekly). Micro-computed tomography and histological evaluation were performed to characterize femoral structures, tooth extraction sockets, and osteonecrosis areas. The results showed that ZOL/Dex significantly increased bone mass compared to saline/Dex, while the simultaneous administration of LP and ZOL/Dex diminished the ZOL-induced enhancement of bone mass. In the alveolar bone around the tooth extraction socket, necrotic bone was significantly increased in the LP/Dex group compared to the saline/Dex group. However, no signs of more severe ONJ-like lesions were observed following combined administration of LP and ZOL/Dex, other than an increase in the number of non-attached TRAP-positive cells. Our findings in a mouse model suggest that LP use can be a risk factor for the development of ONJ.

Functional regulation of osteoblastic MC3T3E-1 cells by hyperbaric oxygen treatment.

OBJECTIVES: The purpose of the present study was to examine the influence of hyperbaric oxygen (HBO) on the function of osteoblastic MC3T3-E1 cells. DESIGN: Murine MC3T3-E1 cells were exposed to HBO treatment (at 2.5 absolute atmospheric pressure with 100% oxygen, 90 min per day) for 28 days. Alkaline phosphatase (ALP) staining, activity, and calcium (Ca) content were measured. Gene expression of vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF), hypoxia-inducible factor-1α (HIF-1α), type 1 collagen (COL1), and osteocalcin (OCN) was assessed using real-time quantitative polymerase chain reaction after a single HBO exposure for 1.5, 6, and 12 h. Furthermore, adenosine triphosphate (ATP) levels were measured using a luminescent cell viability assay. RESULTS: ALP activity and Ca content were higher in the HBO group compared to those in the control group. Gene expression of bFGF, COL1, and OCN was upregulated in the HBO group; however, that of VEGF and HIF-1α significantly decreased in the HBO group in comparison with that in the control group. ATP levels were significantly higher in the HBO group compared to those in the control group. CONCLUSIONS: These findings suggest that HBO accelerates bone formation by increasing the ATP levels of osteoblasts, and bFGF can act as a substitute for VEGF in vascularization by HBO application.

Generalized periodontitis treated with periodontal, orthodontic, and prosthodontic therapy: A case report.

BACKGROUND: Generalized periodontitis is a severe periodontal disease characterized by rapid periodontal destruction in healthy persons. This case report describes the treatment of a severe crowding, large overjet, and occlusal collapse due to the loss of anterior guidance with generalized periodontitis. CASE SUMMARY: A 35-year-old female patient with a chief complaint of crowding and maxillary protrusion was diagnosed with generalized periodontitis by clinical and radiographic examinations. To improve crowding and overjet, orthodontic treatment was performed after basic periodontal therapy. Severely damaged upper right lateral incisor and left canine were extracted, and lower right first premolar and left second premolar were also removed to treat severe crowding. After orthodontic treatment, periodontal flap surgery for upper left molars and guided tissue regeneration for the lower left second molar was performed. Then, a dental implant was inserted in the upper left canine legion. The esthetics of the maxillary anterior tooth was improved by prosthetic restorations. The treatment result showed a well-improved occlusion with proper anterior guidance and healthy periodontal tissue after a retention period of 10 years. CONCLUSION: Periodontal, orthodontic, and prosthodontic treatments are extremely useful to improve function and stable periodontal tissue for generalized periodontitis.

Changes in tongue-palatal contact during swallowing in patients with skeletal mandibular prognathism after orthognathic surgery.

This study aimed to evaluate improvement of tongue-palatal contact patterns during swallowing after orthognathic surgery in mandibular prognathism patients. Thirty patients with mandibular prognathism treated by orthognathic surgery (average age of 27 years, 3 months) and 10 controls (average age 29 years, 6 months) participated in this study. Tongue-palatal contact patterns of patients before and three months after surgery were evaluated by electropalatography (EPG) as well as controls. Whole total of tongue-palatal contact at 0.3, 0.2, and 0.1 sec before complete tongue-palatal contact during swallowing were evaluated. The duration of swallowing phases was also examined. Complete contact of tongue-tip in the alveolar part of individual artificial EPG plate were shown at 0.3, 0.2, and 0.1 sec before complete tongue-palatal contact in the controls, although incomplete contact in the alveolar part were shown at 0.3 sec in mandibular prognathism patients. Whole total of tongue-palatal contact at 0.3 and 0.2 sec before complete tongue-palatal contact was significantly lower in the patients before surgery than in the controls (p<0.05). However, these values increased after surgery. The duration of oral and pharyngeal phase was significantly longer in the patients before surgery than in the controls and the patients after surgery (p<0.01). This study demonstrated that the tongue-palatal contact pattern improved and the duration of oral and pharyngeal phase was shortened in mandibular prognathism patients during swallowing after orthognathic surgery. It is suggested that changes in maxillofacial morphology by orthognathic surgery can induce normal tongue movement during swallowing. (The data underlying this study have been uploaded to figshare and are accessible using the following DOI: https://doi.org/10.6084/m9.figshare.14101616.v1).

Bonding and Thermal Cycling Performances of Two (Poly)Aryl-Ether-Ketone (PAEKs) Materials to an Acrylic Denture Base Resin.

Poly(aryl-ether-ketone) materials (PAEKs) are gaining interest in everyday dental practices because of their natural properties. This study aims to analyze the bonding performance of PAEKs to a denture acrylic. Testing materials were pretreated by grinding, sandblasting, and priming prior to polymerization with the denture acrylic. The surface morphologies were observed using a scanning electron microscope and the surface roughness was measured using atomic force microscopy. The shear bond strength (SBS) values were determined after 0 and 2500 thermal cycles. The obtained data were analyzed using a paired samples t-test and Tukey's honestly significant difference (HSD) test (α = 0.05). The surface characteristics of testing materials after different surface pretreatments showed obvious differences. PAEKs showed lower surface roughness values (0.02-0.03 MPa) than Co-Cr (0.16 MPa) and zirconia (0.22 MPa) after priming and sandblasting treatments (p < 0.05). The SBS values of PAEKs (7.60-8.38 MPa) met the clinical requirements suggested by ISO 10477 (5 MPa). Moreover, PAEKs showed significantly lower SBS reductions (p < 0.05) after thermal cycling fatigue testing compared to Co-Cr and zirconia. Bonding performance is essential for denture materials, and our results demonstrated that PAEKs possess good resistance to thermal cycling fatigue, which is an advantage in clinical applications. The results imply that PAEKs are potential alternative materials for the removable of prosthetic frameworks.

An Investigation of Tongue-Palatal Contact Changes in Patients With Skeletal Mandibular Lateral Deviation After Sagittal Split Ramus Osteotomy.

ABSTRACT: This study aimed to investigate the changes in tongue-palatal contact patterns in patients with mandibular lateral deviation by electropalatography (EPG) before and after sagittal split ramus osteotomy (SSRO). Ten mandibular asymmetry patients who underwent SSRO participated in the study. Tongue-palatal contact patterns for the production of /t/ and /s/ sounds were observed using EPG before surgery and 3 months after surgery, and the changes in EPG pattern were examined. The number of electrode contacts in the 2 vertical columns of the EPG plate was calculated both in the mandibular deviation side and the nondeviation side. The EPG patterns for /t/ and /s/ showed asymmetry before surgery but became normal after surgery. Before surgery, the number of electrode contacts in the 2 vertical columns in the mandibular deviation side was significantly lower than that in the nondeviation side and the normal participants during /t/ and /s/ articulation. However, the number of electrode contacts in the deviation side significantly increased after surgery. This study demonstrated that the tongue-palatal contact patterns for /t/ and /s/ articulation shifted to the direction of mandibular deviation and improved after SSRO.

Effects of hyperbaric oxygen treatment on calvarial bone regeneration in young and adult mice.

OBJECTIVE: The present study aimed to investigate the effects of hyperbaric oxygen (HBO) treatment on calvarial bone regeneration in young and adult mice. METHODS: Calvarial defects of 6.0 mm diameter were created in sixteen 3-week (young) and sixteen 32-week old (adult) mice. The mice were divided into two groups of eight animals each (HBO-treated and control). The 90-min HBO treatment at 2.5 absolute atmospheric pressure and 100 % oxygen was performed for five days a week for 12 weeks. After 2-weeks from the operation, micro-computerized tomography and video microscopy were used to evaluate the regenerated bone volume and microcirculation every two weeks. The protein concentrations of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) in exudates of the calvarial tissue field were measured at 1, 2, 3, and 4 weeks after surgery. After 12 weeks, histochemical examination of regenerated calvarial bone was conducted. RESULTS: Regenerated bone was formed earlier in young mice than in adult mice treated with HBO. HBO stimulates angiogenesis in the periosteum around regenerated bone area in both young and adult mice at 2 weeks. VEGF concentrations in the calvarial tissue field were lower in the HBO group than in the control 1 week after operation, although bFGF were higher till the 2nd week in the HBO group than in the control. CONCLUSIONS: HBO accelerates bone regeneration earlier in young mice than in adult mice. In the HBO-treated group, bFGF expression was promoted at an early stage, although the expression of VEGF was inhibited.

Effect of continuous compressive force on the expression of RANKL, OPG, and VEGF in osteocytes.

The present study aimed to investigate the effect of a compressive force (CF) on the expression of receptor activator of nuclear factor kappa-B ligand (RANKL), osteoprotegerin (OPG), and vascular endothelial growth factor (VEGF) in murine osteocytes (MLO-Y4) as well as animal study. After application of a CF for 1, 3, 6, and 12 h, gene and protein expression of RANKL, OPG, and VEGF in MLO-Y4 cells were determined by real-time PCR and enzyme-linked immunosorbent assay (ELISA). Furthermore, the effect of a stretch-activated (S-A) channel was examined by gadolinium (Gd3+) administration. In an animal experiment, the expression of these factors in osteocytes of alveolar bone was examined after experimental tooth movement in rats. After CF application, significant increases in RANKL, VEGF and RANKL/OPG ratio were shown. The upregulated gene and protein levels of these factors were reduced by Gd3+ administration. After tooth movement, upregulated RANKL and VEGF were imunohistochemically shown in osteocytes of alveolar bone. These findings suggest that CF application on osteocytes elevates expression of osteoclast-inducing factor and angiogenesis factor in vivo and vitro.

Acetaminophen reduces apical root resorption during orthodontic tooth movement in rats.

OBJECTIVE: The present study aimed to investigate the inhibitory effect of acetaminophen on apical root resorption during orthodontic tooth movement by controlling inflammation in the periodontal ligament and apical pulp tissue. METHODS: Human periodontal ligament and pulp cells were subjected to 10 kPa of cyclic tensile force (CTF) in a Flexcell Strain Unit for 48 h. Then, 10 and 100 µM acetaminophen were added to the culture medium, and the expression of interleukin (IL)-1B, receptor activator of nuclear factor kappa-B ligand (RANKL), tumor necrosis factor (TNF)α, and colony stimulating factor 1 (CSF1) were evaluated. In an animal experiment, the upper first molars of 7-week-old rats were moved mesially by applying 10 g of orthodontic force. After 30 days of force application, the effects of acetaminophen on apical root resorption were examined. RESULTS: In both the periodontal ligament and pulp cells, the expression levels of IL-1B, TNFα, RANKL, and CSF1 were significantly higher in the CTF-treated group than in the control group. However, the expression levels of these factors were decreased by acetaminophen administration. High expression of IL-1B, TNFα, RANKL, and CSF1 at the root apex were also detected immunohistochemically in rats after tooth movement, but were decreased by acetaminophen administration. In addition, the number of odontoclasts and the amount of apical root resorption were significantly decreased in the acetaminophen group. Importantly, no significant difference in tooth movement was observed between the acetaminophen and control groups. CONCLUSIONS: These results suggest that acetaminophen can reduce severe root resorption in the apex area without disturbing orthodontic tooth movement.

Influences of mouth breathing on memory and learning ability in growing rats.

This study was to investigate the influences of habitual mouth breathing on memory and learning ability during the growth period. At age 5 weeks, the experimental rats were subjected to surgery to close completely one side of the nasal cavity. An 8-arm radial maze was used to evaluate memory and learning ability at age 7, 11, and 15 weeks. Moreover, the brain was extracted at age 7, 11, and 15 weeks, and subjected to histomorphometric examination for the distribution and number of pyramidal cells in the hippocampal CA1 and CA3 regions after Nissl staining. The trial time to accomplish each task was significantly longer in the experimental group than in the control group throughout the experimental period. The number of pyramidal cells was significantly less in the experimental rats than in controls in both the CA1 and CA3 regions for the entire experimental period. Thus, the functional deterioration of the respiratory system during the growth phase exerts a substantial effect on the growth and development of the central nervous system.

Effects of loxoprofen on the apical root resorption during orthodontic tooth movement in rats.

Studies have revealed that severe apical root resorption during tooth movement is caused by the noninfective inflammatory reaction of apical root tissues. We hypothesized that loxoprofen can suppress apical root resorption during tooth movement. Cyclic tensile force (CTF) of 10 kPa was applied to the human pulp cells for 48 hours by the Flexcell Strain Unit. Loxoprofen (10 and 100 µM) was added to the culture cells, and expression of cyclooxygenase (COX)-1, COX-2, interleukin (IL)-1ß, receptor activator of nuclear factor kappa-B ligand (RANKL), tumor necrosis factor (TNF)-α, and macrophage colony-stimulating factor (M-CSF) were examined. To determine the effects of loxoprofen sodium on apical root reabsorption during tooth movement, the upper first molars of 7-week-old rats were subjected to mesial movement by 10g force for 30 days with or without the oral administration of loxoprofen. Gene expression and protein concentration of COX-1, COX-2, IL-1ß, TNF-α, RANKL and M-CSF were significantly higher in the CTF group than in the control group. However, these levels were decreased by loxoprofen administration. After orthodontic tooth movement, the expression of IL-1ß, TNF-α, RANKL and M-CSF decreased in the loxoprofen group than in the control group by immunohistochemical staining. In comparison to control group, less number of odontoclasts and a decrease in the amount of apical root resorption was observed in the loxoprofen group. Many osteoclasts became visible on the pressure side of the alveolar bone in the both groups, and the amount of tooth movement did not show a significant difference. These findings demonstrate that severe apical root resorption may be suppressed by loxoprofen administration, without a disturbance of tooth movement.

Comparison of the bone regeneration ability between stem cells from human exfoliated deciduous teeth, human dental pulp stem cells and human bone marrow mesenchymal stem cells.

Cleft lip and palate is the most common congenital anomaly in the orofacial region. Autogenous iliac bone graft, in general, has been employed for closing the bone defect at the alveolar cleft. However, such iliac bone graft provides patients with substantial surgical and psychological invasions. Consequently, development of a less invasive method has been highly anticipated. Stem cells from human exfoliated deciduous teeth (SHED) are a major candidate for playing a significant role in tissue engineering and regenerative medicine. The aim of this study was to elucidate the nature of bone regeneration by SHED as compared to that of human dental pulp stem cells (hDPSCs) and bone marrow mesenchymal stem cells (hBMSCs). The stems cells derived from pulp tissues and bone marrow were transplanted with a polylactic-coglycolic acid barrier membrane as a scaffold, for use in bone regeneration in an artificial bone defect of 4 mm in diameter in the calvaria of immunodeficient mice. Three-dimensional analysis using micro CT and histological evaluation were performed. Degree of bone regeneration with SHED relative to the bone defect was almost equivalent to that with hDPSCs and hBMSCs 12 weeks after transplantation. The ratio of new bone formation relative to the pre-created bone defect was not significantly different among groups with SHED, hDPSCs and hBMSCs. In addition, as a result of histological evaluation, SHED produced the largest osteoid and widely distributed collagen fibers compared to hDPSCs and hBMSCs groups. Thus, SHED transplantation exerted bone regeneration ability sufficient for the repair of bone defect. The present study has demonstrated that SHED is one of the best candidate as a cell source for the reconstruction of alveolar cleft due to the bone regeneration ability with less surgical invasion.

RANKL and OPG expression: Jiggling force affects root resorption in rats.

OBJECTIVE: To immunohistochemically investigate the longitudinal changes in root resorption by jiggling force in experimental animal models. MATERIALS AND METHODS: Fifty-six 12-week-old male Wistar rats were used. The maxillary first molars were alternately moved in the buccal and lingual direction in 28 rats (experimental group) using an experimental appliance to produce jiggling forces of 10 g. In another 28 rats (control group), the maxillary first molars were moved in only the lingual direction with a force of 10 g. After 1, 3, 7, 10, 14, 17, and 21 days, the maxillae were resected and subjected to immunohistochemical analysis. The resorption area was quantified histomorphometrically and the number of odontoclasts on the root surface was counted. Expression of RANKL and OPG was also examined by immunohistochemical staining. RESULTS: The root resorption area and the number of odontoclasts were significantly greater in the experimental group than in controls. Odontoclasts were detected in the resorption lacunae and PDL in the experimental group, whereas osteoclasts were located only along the alveolar bone in controls. OPG was detected on the alveolar bone in the experimental group and on the root surfaces of the controls. CONCLUSIONS: Jiggling force is a critical factor in severe root resorption, affecting RANKL and OPG expression, which accelerates and inhibits odontoclastic induction, respectively.

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.

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.

Effects of pulpectomy on the amount of root resorption during orthodontic tooth movement.

INTRODUCTION: Previous studies have revealed that orthodontic force affects dental pulp via the rupture of blood vessels and vacuolization of pulp tissues. We hypothesized that pulp tissues express inflammatory cytokines and regulators of odontoclast differentiation after excess orthodontic force. The purpose of this study was to investigate the effects of tensile force in human pulp cells and to measure inflammatory root resorption during tooth movement in pulpless rat teeth. METHODS: After cyclic tensile force application in human pulp cells, gene expression and protein concentration of macrophage colony-stimulating factor, receptor activator of nuclear factor kappa-B ligand, interleukin-1 beta, and tumor necrosis factor alpha were determined by real-time polymerase chain reaction and enzyme-linked immunoassay. Moreover, the role of the stretch-activated channel was evaluated by gadolinium (Gd(3+)) treatment. The upper right first molars of 7-week Wistar rats were subjected to pulpectomy and root canal filling followed by mesial movement for 6 months. RESULTS: The expression of cytokine messenger RNAs and proteins in the experimental group peaked with loading at 10-kPa tensile force after 48 hours (P < .01). Gd(3+) reduced the expression of these cytokine messenger RNAs and protein concentrations (P < .01). The amount of inflammatory root resorption was significantly larger in the control teeth than the pulpectomized teeth (P < .05). CONCLUSIONS: This study shows that tensile forces in the pulp cells enhance the expression of various cytokines via the S-A channel, which may lead to inflammatory root resorption during tooth movement. It also suggests that root canal treatment is effective for progressive severe inflammatory root resorption during tooth movement.

Cranial bone regeneration after cranioplasty using cryopreserved autogenous bone by a programmed freezer with a magnetic field in rats.

BACKGROUND: The purpose of this study was to develop a bone tissue bank using a programmed freezer with a magnetic field. Parietal bones were removed from rats and used for organ culture examination (non-cryopreserved, cryopreserved with a magnetic field (CAS) and cryopreserved without a magnetic field group). Next, other parietal bones were used for histological examination. The cryopreserved bones by a CAS freezer and dried bones were transplanted respectively. Control bones were replanted without cryopreservation. Animals were sacrificed at 4, 8, 12 and 24 weeks after surgery. After organ culture, the isolated osteoblasts from parietal bones which were cryopreserved by a CAS freezer can survive and proliferate as much as non-cryopreserved group. Histological examinations showed new bone formation in control and CAS group. These results suggest that bone tissue cryopreservation by CAS freezer can be successfully used for bone grafting which may be a novel option for regeneration medicine.

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.