The origin of lumbar subcutaneous edema: two case reports.

Since magnetic resonance imaging (MRI) is widely used to evaluate complaints of low back pain, there have been many reports of lumbar subcutaneous edema (LSE). However, the mechanism underlying its development is unknown. We herein report 2 cases that showed the reduction of LSE. These cases suggest details concerning the mechanism underlying the development of LSE. The first case was an obese 70-year-old woman with a history of chronic back pain due to lumbar canal stenosis. MRI revealed LSE extending from the level of the L2 vertebral body to the sacrum. However, LSE was reduced following weight loss due to a stomach ulcer. This case clearly indicated obesity as the cause of LSE. The second case was a nonobese 31-year-old woman with acute excruciating low back pain due to thoracolumbar fascia strain. LSE was observed at the level of the L3-L4 vertebral body. Two weeks later, her low back pain and LSE were reduced. This case suggests that the origin of LSE was impairment of the thoracolumbar fascia due to strain. We hypothesize that the mechanism underlying the development of LSE may be lymphatic or interstitial fluid pooling due to disturbance of the lumbar fascia.

Effects of Neodymium-Doped Yttrium Aluminium Garnet (Nd:YAG) Laser Irradiation on Bone Metabolism During Tooth Movement.

INTRODUCTION: The aim of this study is to evaluate the effects of low-level neodymium-doped yttrium aluminium garnet (Nd:YAG) laser irradiation on orthodontic tooth movement and histological examination. METHODS: Eleven male Wistar rats (aged 10 weeks) were included. To produce experimental tooth movement in rats, 10 g force was applied to maxillary first molars with nickel titanium closed coil springs. Right molars were irradiated with Nd:YAG laser on days 0, 1, 2, 3, 7, 10, 14, 17, 21 and 24, while un-irradiated left molars were used as control. Distance between mesial side of second molar and distal side of first molar was measured on µCT image during tooth movement and the rats were sacrificed 4 weeks after the initiation of tooth movement. RESULTS: The amount of tooth movement was significantly greater in the irradiation group (0.20 ± 0.06) than in the control group (0.14 ± 0.03) during the first week (P < 0.05). However, no statistically significant difference was found afterwards. There was a tendency of higher tartrate-resistant acid phosphatase (TRAP)-positive nuclei count in the pressure zones of the laser irradiation group, but it was not statistically significant. In immuno-histological examination, expressions of alkaline phosphatase (ALP) and receptor activator of nuclear factor kappa-B ligand (RANKL) were higher at the pressure site of the laser irradiation group than the control group, whereas there was no difference in osteoprotegerin (OPG) expression. CONCLUSION: The results suggest that low-level Nd:YAG laser may stimulate osteoclast and osteoblast activation and accelerate bone metabolism during tooth movement.

Role of articular disc in condylar regeneration of the mandible.

The articular disc in the temporomandibular joint plays an important role in mandibular growth. Functional appliances induce regeneration of the condyle even after condylectomy. The aim of this study was to examine the role of the articular disc in regeneration of the condyle after unilateral condylectomy with use of a functional appliance in growing rats. Fifty growing rats were subjected to unilateral condylectomy and then half of them underwent discectomy. The functional appliance was applied to half of the rats in each group to induce regeneration of the condyle. Four and eight weeks later, morphometric and histologic analyses of the mandible were performed. Regeneration of the condyle was demonstrated in the two condylectomy groups. In the condylectomy+appliance group, the shape and cartilage of the condyle were equivalent to a normal condyle. However, regeneration of the condyle was not observed in the two discectomy groups even with the use of the functional appliance. The articular disc appears to be crucial in the regeneration of a damaged condyle, suggesting that defects or damage to the articular disc may influence mandibular growth and regeneration or repair of the condyle.

Factors related to stability following the surgical correction of skeletal open bite.

OBJECTIVES: If a skeletal anterior open bite malocclusion is treated by orthognathic surgery directed only at the mandible, the lower jaw is repositioned upward in a counter-clockwise rotation. However, this procedure has a high risk of relapse. In the present study, the key factors associated with post-surgical stability of corrected skeletal anterior open bite malocclusions were investigated. MATERIAL AND METHODS: Eighteen orthognathic patients were subjected to cephalometric analysis to assess the dental and skeletal changes following mandibular surgery for the correction of an anterior open bite. The patients were divided into two groups, determined by an increase or decrease in nasion-menton (N-Me) distance as a consequence of surgery. Changes in overbite, the displacements of molars and positional changes in Menton were evaluated immediately before and after surgery and after a minimum of one year post-operatively. RESULTS: The group with a decreased N-Me distance exhibited a significantly greater backward positioning of the mandible. The group with an increased N-Me distance experienced significantly greater dentoalveolar extrusion of the lower molars. CONCLUSIONS: A sufficient mandibular backward repositioning is an effective technique in the prevention of open bite relapse. In addition, it is important not to induce molar extrusion during post-surgical orthodontic treatment to preserve stability of the surgical open bite correction.

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.

Changes in the lip-line in asymmetrical cases treated with isolated mandibular surgery.

OBJECTIVE: The aim of this study was to investigate changes in the lip-line in asymmetrical cases treated with mandibular osteotomy alone. DESIGN: Retrospective study. SETTING: Hiroshima University Institute of Biomedical & Health Sciences, Hiroshima, Japan METHODS: The subjects in this study consisted of 30 patients with an altered lip-line inclination who underwent isolated mandibular osteotomy (bilateral or unilateral sagittal split ramus osteotomy or intraoral vertical ramus osteotomy) as part of their surgical correction. Frontal cephalograms and facial photographs, taken at the first examination and after treatment, were used to measure changes in the inclination of the lip-line and cant of the occlusal plane, as well as the lateral deviations of hard and soft tissue Menton. RESULTS: Inclination of the lip-line after active treatment was significantly improved compared with that before treatment. Menton on the hard and soft tissues after active treatment also experienced a significant improvement compared with its position at first examination. CONCLUSIONS: One-jaw mandibular osteotomy is able to improve the inclination of the lip-line even in the presence of an occlusal cant. The inclination of the lip-line is corrected in association with sufficient lateral movements of Menton on the mandible.

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.

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.

Gummy smile and facial profile correction using miniscrew anchorage.

This case report describes the treatment of a case involving a skeletal Class II facial profile with a gummy smile. While treating a facial profile and a gummy smile, the outcome may not always be successful with orthodontic therapy alone. For this reason, surgical therapy is often chosen to gain an esthetic facial profile and a good smile. However, sometimes the patients reject surgical treatment and an alternative method must be considered. Skeletal anchorage systems such as miniscrews are now frequently used for correcting severe malocclusion that should be treated by surgical therapy. In this case report, we treated a skeletal Class II malocclusion with a convex profile and a gummy smile using miniscrews, which were placed in the upper posterior and anterior areas. The active treatment period was 3.5 years, and the patient's teeth continued to be stable after a retention period of 36 months.

Age-related production of osteoclasts and the changes of serum levels of vascular endothelial growth factor (VEGF) and receptor activator for nuclear factor (NF)-κB ligand (RANKL) in osteopetrotic (op/op) mice.

OBJECTIVE: Expression of osteoclasts in osteopetrotic (op/op) mice is substantially reduced by the absence of functional macrophage colony-stimulating factor (M-CSF). However, it has been reported that osteoclasts do gradually appear in the bones of op/op mice and spontaneously correct the osteopetrosis. DESIGN: Age-related production of osteoclasts and the changes of serum levels of vascular endothelial growth factor (VEGF) and receptor activator for nuclear factor (NF)-κB ligand (RANKL) in op/op mice were examined. RESULTS: The number of femoral osteoclasts, and the serum levels of VEGF, both gradually increased in op/op mice after birth and reached a peak in 120- and 60-day-old mice, respectively. However, the serum levels of RANKL showed an inverse relationship to osteoclast number. CONCLUSIONS: These findings suggest that the appearance of osteoclasts may be influenced by the serum levels of VEGF and that the serum levels of RANKL may be influenced by the appearance of osteoclasts.

Uprighting of severely impacted mandibular second molars: a case report.

INTRODUCTION: The incidence of mandibular first and second molar impaction is increasing but still recorded as rare. Treatment methods involving uprighting, extraction, or autologous tooth transplantation have been described. AIM: The present study describes the uprighting of 3 impacted mandibular second molars presenting with eruptive disorders. METHODS: The application of limited and appropriate orthodontic therapy completed treatment in 11 months, 5 months, and 2 years and 3 months, respectively. Although no absolute anchorage in the form of miniscrews was required, no significant anchorage demands were considered necessary. Although the third molar tooth germs were identified and preserved in each case, no adverse influence on the uprighting of the second molars was encountered. RESULTS: The favourable molar repositioning results were likely due to the youth of the 3 patients as the third molars were in early development and bone remodelling was marked. Furthermore, no problems related to anchorage or alveolar bone loss were identified after treatment. CONCLUSION: The results indicated the benefits of limited orthodontic treatment and early intervention for the uprighting of impacted mandibular second molars.

In-vitro and in-vivo study of periodontal ligament cryopreserved with a magnetic field.

INTRODUCTION: The purpose of this study was to examine the effect of a new cryopreservation method with a magnetic field on periodontal regeneration in vitro and in vivo. METHODS: Human periodontal ligament cells were frozen in 10% dimethyl sulfoxide by using a programmed freezer with a magnetic field. Cells were cryopreserved for 3 days at -150°C. Immediately after thawing, collagen type I and alkaline phosphatase gene expression were determined by real-time polymerase chain reaction. Incisors were extracted from 15-week-old Wistar rats and cryopreserved or dried for 3 days. Then the incisors were replanted into the same sockets. Ninety days after transplantation, they were observed under light microscopy. RESULTS: There was no difference in the messenger RNA expression of collagen type I between the cryopreserved and the control groups. The expression of alkaline phosphatase messenger RNA in the cryopreserved group was slightly decreased compared with the control group. There was no progressive root resorption in the teeth that were replanted immediately (control group) or cryopreserved. However, there was widespread root resorption and ankylosis in the dried teeth. CONCLUSIONS: These results show that a magnetic field programmed freezer can be successfully used for cryopreservation of teeth.

Treatment of a case of skeletal class II malocclusion with temporomandibular joint disorder using miniscrew anchorage.

At the present time, there are no reports in the literature on the treatment of temporomandibular joint disorder (TMD) by intrusion of molars using mini-screws. This case report describes the treatment for a female patient, aged 19 years seven months, with a TMD and an excessive lower anterior facial height. Overjet and overbite were +5.0 mm and +0.5 mm, respectively. The patient had a history of orthodontic treatment in which her first premolars were all extracted. During the first orthodontic treatment, a clockwise mandibular rotation was observed as a result of the increase of posterior dentoalveolar height. She had temporomandibular joint (TMJ) pain during mouth opening and complained of difficulty in eating due to masticatory dysfunction. The pretreatment Schuller views of both TMJ showed a posterior condyle position. In order to correct the overjet, molar relationship and the mandibular condyle position, a miniscrew was inserted into the palatal region of the upper first molar to intrude the upper posterior teeth. As the upper molars were intruded, the overjet was decreased, and a class I molar relationship was achieved by a counterclockwise mandibular rotation. After one year of treatment, an acceptable occlusion was achieved, and the condyle moved into centric position in the glenoid fossa. The patient's teeth continued to be stable, and she had no pain in TMJ after a retention period of three years. The result of this treatment showed that molar intrusion using miniscrew anchorage is effective for treatment of a TMD patient with a posterior condyle position.

Changes in cortical bone mineralization in the mouse mandible with regenerated condyle.

The objective of this study was to evaluate the effects of condylectomy and of a functional appliance on the mineral content, density, and area of the mandibular cortical bone in a growing mouse model. In the condylectomy group, a unilateral condylectomy was performed on the right side of the mouse mandible. In the condylectomy + appliance group, a functional appliance was used to reposition the mandible after the unilateral condylectomy. All mice were killed 4 wk after surgery. Each mandible was then subjected to analyses of cortical bone mineral content (CRT_CNT), cortical bone density (CRT_DEN), and cortical bone area (CRT_A) by peripheral quantitative computed tomography (pQCT). The CRT_CNT, CRT_DEN, and CRT_A values were significantly lower in the condylectomized mandible (right side) than in the non-condylectomized mandible (left side). However, in the condylectomized animals in which a functional appliance was used, the CRT_CNT, CRT_DEN, and CRT_A values became higher than those in the mice treated with condylectomy alone. No significant differences were found in the CRT_CNT, CRT_DEN, and CRT_A between non-condylectomized (left side) mandibles, mandibles treated with condylectomy + functional appliance (right side), and control mandibles. It was thus shown that a functional appliance used to reposition the condylectomized mandible forward in a symmetric position induced improvement of the cortical bone in a mouse model in terms of cortical bone mineral content, density, and area.

Mesenchymal stem cell-induced cranial suture-like gap in rats.

BACKGROUND: Mesenchymal stem cells are used in the reconstruction of many organs and tissues. However, there are no data regarding cranial suture regeneration using mesenchymal stem cells. Because cranial sutures are important for cranial bone growth, the authors examined whether mesenchymal stem cells could aid with suture reformation in adolescent rats. METHODS: Mesenchymal stem cells were isolated from bone marrow of rat femora. Twenty 4-week-old Fischer 344 male rats received sagittal suture and bone defects with a diameter of 6.0 mm, and mesenchymal stem cells were grafted with membranes. Twenty control rats underwent empty membrane transplantation. The mesenchymal stem cell and control groups were killed at 4, 8, 16, and 24 weeks after surgery. The parietal bones, including the sagittal suture, were observed under a light microscope. Bone structures were measured on digitized photomicrographs captured in a computer. For each sample, bone and suture regeneration were observed by dorsoventral cephalograms. RESULTS: In 4- and 8-week control and mesenchymal stem cell groups, a small volume of new bone formation was observed. In the 16- and 24-week mesenchymal stem cell groups, a large amount of new bone formation and a suture-like gap were identified. In contrast, only a small volume of new bone formation along with craniosynostosis was detected in the 24-week control group. CONCLUSION: These results suggest that mesenchymal stem cell grafting may be a novel option for cranioplasty, not only repairing bone tissue but also resulting in suture-like gap formation, which may advance cranial bone growth.

Expression of Sox 9 and type II and X collagens in regenerated condyle.

The present study was designed to examine the expression of Sox 9 and type II and X collagens in regenerated condyle resulting from the use of a functional appliance. Ninety, 3-week-old, mice were divided equally into the following groups: two experimental groups (condylectomy group and condylectomy with functional appliance group) and the corresponding control group. In the condylectomy group, a unilateral condylectomy was performed on the right side. In the condylectomy with appliance group, the mandible was repositioned in a forward direction using a functional appliance after unilateral condylectomy. The expression of Sox 9 and type II and X collagens in the condyle was determined immunohistochemically 4 weeks after surgery. In mice with a condylectomy, the expression was minimal. On the other hand, these factors were highly expressed in the condylectomized side with the appliance. It is thus speculated that cartilaginous regeneration is due to the expression of chondrogenic factors, such as Sox 9 and type II and X collagens. It is also suggested that condyle regeneration results from an optimal intra-articular environment with appropriate joint spaces achieved by condylar repositioning.

A newly developed snack effective for enhancing bone volume.

BACKGROUND: The incidence of primary osteoporosis is higher in Japan than in USA and European countries. Recently, the importance of preventive medicine has been gradually recognized in the field of orthopaedic surgery with a concept that peak bone mass should be increased in childhood as much as possible for the prevention of osteoporosis. Under such background, we have developed a new bean snack with an aim to improve bone volume loss. In this study, we examined the effects of a newly developed snack on bone volume and density in osteoporosis model mice. METHODS: Orchiectomy (ORX) and ovariectomy (OVX) were performed for C57BL/6J mice of twelve-week-old (Jackson Laboratory, Bar Harbar, ME, USA) were used in this experiment. We prepared and given three types of powder diet e.g.: normal calcium diet (NCD, Ca: 0.9%, Clea Japan Co., Tokyo, Japan), low calcium diet (LCD, Ca: 0.63%, Clea Japan Co.,) and special diet (SCD, Ca: 0.9%). Eighteen weeks after surgery, all the animals were sacrified and prepared for histomorphometric analysis to quantify bone density and bone mineral content. RESULTS: As a result of histomorphometric examination, SCD was revealed to enhance bone volume irrespective of age and sex. The bone density was increased significantly in osteoporosis model mice fed the newly developmental snack as compared with the control mice. The bone mineral content was also enhanced significantly. These phenomena were revealed in both sexes. CONCLUSION: It is shown that the newly developed bean snack is highly effective for the improvement of bone volume loss irrespective of sex. We demonstrated that newly developmental snack supplements may be a useful preventive measure for Japanese whose bone mineral density values are less than the ideal condition.

Inhibitory effects of constituents of Morinda citrifolia seeds on elastase and tyrosinase.

A 50% ethanolic extract (MCS-ext) from seeds of Morinda citrifolia ("noni" seeds) showed more potent in vitro inhibition of elastase and tyrosinase, and 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity than extracts of M. citrifolia leaves or flesh. Activity-guided fractionation of MCS-ext using in vitro assays led to the isolation of ursolic acid as an active constituent of elastase inhibitory activity. 3,3'-Bisdemethylpinoresinol, americanin A, and quercetin were isolated as active constituents having both tyrosinase inhibitory and radical scavenging activities. Americanin A and quercetin also showed superoxide dismutase (SOD)-like activity. These active compounds were isolated from noni seeds for the first time.

Fms-like tyrosine kinase (Flt)-4 signaling participates in osteoclast differentiation in osteopetrotic (op/op) mice.

Vascular endothelial growth factor (VEGF) induces osteoclast differentiation as well as neovascularization by binding to the fms-like tyrosine kinase (Flt)-1 and fetal liver kinase (Flk)-1 receptors. The Flt-4 receptor also plays an important role in angiogenesis and lymphangiogenesis. The purpose of this study was to investigate the functions of Flt-4 in the signaling pathway of osteoclast differentiation. We examined the expression of Flt-4 on osteoclast precursor cells (OCPs), and the ability of recombinant human (rh) VEGF-D, one of the ligands of Flt-4, to stimulate the phosphorylation of extracellular-regulated kinase1/2 (ERK1/2) and to activate the nuclear factor-kappa B (NF-kappaB) pathway in OCPs. The number of osteoclasts induced by injection of rhVEGF-D in osteopetrotic (op/op) mice was also evaluated in the absence or presence of neutralizing antibodies to Flt-4. Flt-4 expression was detected on OCPs at both gene and protein levels and stimulation of Flt-4 by rhVEGF-D might induce activation of mitogen-activated protein kinase (MAPK) and NF-kappaB pathways for induction of osteoclast differentiation. Moreover, the number of osteoclasts in op/op mice increased after injection of rhVEGF-D, but was significantly reduced by the injection of Flt-4 neutralizing antibodies. We have therefore shown that Flt-4 expressed on OCPs, might activate MAPK and NF-kappaB pathways and played an important role in osteoclast differentiation.