Distribution of hemokinin-1 in the rat trigeminal ganglion and trigeminal sensory nuclear complex.

OBJECTIVE: A new mammalian tachykinin peptide encoded in a TAC4 gene was identified and designated as hemokinin-1 (HK-1). A representative of the tachykinin peptide family is substance P (SP), and the function of SP has been well characterized as a pain transmitter or modulator, while it is possible that HK-1 is involved in pruriceptive processing, but, as yet, the distribution of HK-1 peptide in the trigeminal sensory system is still unknown. Thus, the aim of the present study was to elucidate the distribution of HK-1, while comparing the expression of SP, in the trigeminal ganglion and trigeminal sensory nuclear complex. DESIGN: The trigeminal ganglion and the brain stem of male SD rats were used in the immunohistochemical study. Since the amino acid sequence in the carboxyl-terminal regions of HK-1 and SP is common, polyclonal antibodies of HK-1 and SP derived from 6 amino acids consisting of amino-terminal regions of these peptides were produced in guinea pig and rabbit, respectively. The immunohistochemical staining of HK-1 and SP was conducted using frozen sections of the trigeminal ganglion and brain stem in rats. RESULTS: Immunohistochemical studies revealed the expression of HK-1 in small- and medium-sized trigeminal ganglion neurons, in the paratrigeminal nucleus, and in lamina I of the trigeminal nucleus caudalis, while there was no immunoreactivity of HK-1 in the trigeminal nucleus principalis, trigeminal nucleus oralis, and trigeminal nucleus interpolaris. CONCLUSION: These findings indicate that HK-1 is a target molecule for treatment of itch in the orofaicial regions.

Development of a complete human anti-human transferrin receptor C antibody as a novel marker of oral dysplasia and oral cancer.

Oral squamous cell carcinoma (OSCC) is the sixth most common cancer worldwide. Up to 20% of oral dysplasia cases have been suggested to undergo malignant transformation to OSCC; however, there are no methods to predict OSCC development. In this study, to identify the genes associated with oral dysplasia progression, we performed genomic copy number analyses of genomic DNA samples isolated from primary oral dysplasia and OSCC via the microdissection method and found elevated expression of transferrin receptor C (TfR1/TFRC) with genomic amplification in oral dysplasia and OSCC. The expression rate of TFRC in OSCC was significantly higher than that in dysplasia, suggesting that OSCC disease progression might be related to TFRC expression. Additionally, we investigated the in vitro and in vivo impacts of a newly established anti-human TFRC monoclonal antibody, which was isolated from a human cDNA library using the phage-display method, on cell proliferation and survival. The anti-TFRC antibody blocked the interaction between transferrin and TFRC and consequently inhibited iron uptake, leading to the iron deprivation-mediated suppression of cell growth and induction of apoptosis. Moreover, we demonstrated that the anti-TFRC antibody efficiently inhibited tumor growth in a murine xenograft OSCC model. Therefore, we suggest our developed complete human anti-human TFRC antibody as a useful, novel treatment for oral dysplasia and OSCC.

The role of spinal serotonin receptor and alpha adrenoceptor on the antiallodynic effects induced by intrathecal milnacipran in chronic constriction injury rats.

Milnacipran, a reuptake inhibitor of noradrenaline (NA) and serotonin (5-HT), elicits an antiallodynic effect in rats with neuropathic pain; however, the role of NA and 5-HT receptors in the induction of the antiallodynic effect of milnacipran remains unclear. Thus, we examined the effects of prazosin as an α1 adrenoceptor antagonist, yohimbine as an α2 adrenoceptor antagonist, metergoline as a 5-HT1, 5-HT2 and 5-HT7 receptor antagonist, cyanopindolol as a 5-HT1A/1B receptor antagonist, ketanserin as a 5-HT2 receptor antagonist, and ondansetoron as a 5-HT3 receptor antagonist on the antiallodynic effect of milnacipran in neuropathic rats with chronic constriction injury (CCI). The CCI rats expressed mechanical and thermal allodynia, which was attenuated by intrathecal injection of milnacipran. Yohimbine, but not prazosin, reversed the milnacipran-induced antiallodynic effect. The antiallodynic effect of milnacipran was also reversed by metergoline, ketanserin and ondansetron, while cyanopindolol reversed the antiallodynic effect on mechanical, but not thermal stimulation. Furthermore, c-Fos expression in lamina I/II of the spinal dorsal horn was enhanced by thermal stimulation and the enhanced expression of c-Fos was suppressed by milnacipran. This effect of milnacipran was reversed by yohimbine, metergoline, katanserin and ondansetron, but not prazosin. These results indicate that the effect of milnacipran on mechanical and thermal allodynia and c-Fos expression is elicited through the α2 adrenoceptor, but not α1 adrenoceptor, and 5-HT2 and 5-HT3 receptors; furthermore, the 5-HT1A/1B receptor is involved in mechanical allodynia, but not thermal allodynia.

Risk factors for breakage of biodegradable plate systems after bilateral sagittal split mandibular setback surgery.

The aim of this retrospective study was to evaluate the risk factors associated with breakage of biodegradable plate systems after bilateral sagittal split mandibular setback. We studied 169 Japanese adults (62 men, 107 women; age range 16-53 years) with deformities of the jaw diagnosed as mandibular prognathism. All patients were treated by bilateral sagittal split osteotomy (BSSO) with 2 biodegradable fixation plates and screws at the anterior mandibular ramus. We collected the following data from the medical records and radiological findings: sex; age; degree of setback; presence of asymmetry; presence of open bite; operation; design of the plate; operating time; and blood loss. Multiple logistic regression analysis was used to find the factors that were independently associated with the dependent variable: breakage of the biodegradable plate system. In 10 of the 169 patients (6%) the biodegradable plate system for the BSSO broke. Factors that influenced whether or not the biodegradable plate system fractured were if they were asymmetrical (odds ratio (OR) 5.35; P=0.02) and had an open bite (OR 5.20; P=0.02). Asymmetry or open bite was significantly associated with breaks in the biodegradable plate system. Biodegradable plates should be used only when loading is minimal.

Criteria and limitations for selecting a sagittal split ramus osteotomy for patients with skeletal mandibular prognathism and open bite.

PURPOSE: The purpose of this study is to elucidate the criteria and limitations for selecting a sagittal split ramus osteotomy (SSRO) alone for patients with skeletal mandibular prognathism and open bite. STUDY DESIGN: The relationship between the mandibular relapse after 1 year and the extent of vertical movement following SSRO alone was investigated for patients with skeletal mandibular prognathism and open bite and the criteria were determined. RESULTS: The limitation in SSRO alone was found to be 3-mm vertical movement at the gonion. There was a significant difference in the mandibular relapse between the patients with >3-mm change and <3-mm change. CONCLUSIONS: For patients with skeletal mandibular prognathism and open bite, if the vertical movement at the gonion is <3 mm, SSRO alone could be an effective treatment. However, if the movement is >3 mm, SSRO alone should be avoided and bimaxillary osteotomies may be a preferable treatment.

Epidermal choristoma arising on the midline gingiva as a congenital epulis: a case report.

We report an extremely rare case of epidermal choristoma in the midline of the maxillary gingiva. A 2-month-old Japanese boy presented with a polypoid mass in the midline of the maxillary gingiva. The initial clinical diagnosis was congenital epulis. Microscopic examination revealed a granular cell layer and melanin pigmentation within the basal cell layer. Furthermore, sebaceous glands and hair follicles were observed within the connective tissue. The histological diagnosis was therefore epidermal choristoma, based on clinical microscopic observations.

Overexpression of the DNA sensor proteins, absent in melanoma 2 and interferon-inducible 16, contributes to tumorigenesis of oral squamous cell carcinoma with p53 inactivation.

The development of oral squamous cell carcinoma (OSCC) is a multistep process that requires the accumulation of genetic alterations. To identify genes responsible for OSCC development, we performed high-density single nucleotide polymorphism array analysis and genome-wide gene expression profiling on OSCC tumors. These analyses indicated that the absent in melanoma 2 (AIM2) gene and the interferon-inducible gene 16 (IFI16) mapped to the hematopoietic interferon-inducible nuclear proteins. The 200-amino-acid repeat gene cluster in the amplified region of chromosome 1q23 is overexpressed in OSCC. Both AIM2 and IFI16 are cytoplasmic double-stranded DNA sensors for innate immunity and act as tumor suppressors in several human cancers. Knockdown of AIM2 or IFI16 in OSCC cells results in the suppression of cell growth and apoptosis, accompanied by the downregulation of nuclear factor kappa-light-chain-enhancer of activated B cells activation. Because all OSCC cell lines have reduced p53 activity, wild-type p53 was introduced in p53-deficient OSCC cells. The expression of wild-type p53 suppressed cell growth and induced apoptosis via suppression of nuclear factor kappa-light-chain-enhancer of activated B cells activity. Finally, the co-expression of AIM2 and IFI16 significantly enhanced cell growth in p53-deficient cells; in contrast, the expression of AIM2 and/or IFI16 in cells bearing wild-type p53 suppressed cell growth. Moreover, AIM2 and IFI16 synergistically enhanced nuclear factor kappa-light-chain-enhancer of activated B cells signaling in p53-deficient cells. Thus, expression of AIM2 and IFI16 may have oncogenic activities in the OSCC cells that have inactivated the p53 system.

Loss of membrane-bound serine protease inhibitor HAI-1 induces oral squamous cell carcinoma cells' invasiveness.

A loss of balance between cell membrane-associated proteases and their inhibitors may underlie cancer invasion and metastasis. We analysed the roles of a membrane- associated serine protease inhibitor, HAI-1, in oral squamous cell carcinoma (OSCC). While membranous HAI-1 was widely observed in cancer cells of human OSCC tissues, this was significantly reduced at the infiltrative invasion front. In vitro, HAI-1 was detected in all eight OSCC cell lines examined, in which its cognate membrane protease, matriptase was also expressed. HAI-1 expression knock-down (KD) in OSCC lines, SAS and HSC-3, reduced the growth of both lines in vitro but significantly enhanced SAS tumourigenicity in vivo, which was accompanied by histological changes suggestive of the epithelial-mesenchymal transition. Both HAI-1-KD lines also exhibited significantly enhanced migratory capability, and membrane-associated but not truncated HAI-1 was required to rescue this phenotype. Other OSCC lines (HSC-2, Sa3, Ca9-22) also showed enhanced migration in response to HAI-1 KD. The enhanced migration is partly attributed to dysregulation of matriptase, as simultaneous matriptase KD alleviated the migration of HAI-1-KD cells. HAI-1 deficiency also altered the expression of CD24, S100A4, CCND2 and DUSP6, all of which are involved in tumour progression. While matriptase was involved in the increased CD24 expression associated with HAI-1 deficiency, the protease appeared to be not responsible for the altered expression of other genes. Therefore, a matriptase-independent mechanism for the invasiveness associated with HAI-1 KD is also present. Together, these observations suggest that HAI-1 has a crucial suppressive role in OSCC cell invasiveness.

Activation of macrophage-stimulating protein by human airway trypsin-like protease.

Macrophage-stimulating protein (MSP) circulates as a proform protein and requires proteolytic processing for activation. Respiratory ciliated cells express the MSP receptor, recepteur d'origine nantais (RON), at the apical surface, which reportedly has an important role in ciliary function. Like RON, human airway trypsin-like protease (HAT) is also expressed at the apical surface of ciliated cells. Here we show that HAT cleaves proMSP at the physiological activation site, Arg483-Val484. MSP processed by HAT could induce chemotactic responses and morphological changes of peritoneal macrophages. In human respiratory epithelial cells, knock down of HAT expression reduced proMSP processing and RON autophosphorylation. We suggest that HAT is important for MSP-RON signaling in the respiratory tract.

Effect of bone quality and position of the inferior alveolar nerve canal in continuous, long-term, neurosensory disturbance after sagittal split ramus osteotomy.

OBJECTIVES: To examine the relationship between the anatomical position and the bone quality of the inferior alveolar nerve (IFAN) canal and long-term neurosensory disturbance (NSD) of the IFAN after a sagittal split ramus osteotomy (SSRO). MATERIAL AND METHODS: CT images were taken of patients with mandibular prognathism. The location of the IFAN and the bone density around the IFAN were measured on CT images. Whether NSD at 6 months and 1 year after SSRO was related to the position and bone quality of the IFAN canal was analyzed. RESULTS: Significant correlations were found between the anatomical position and the bone quality of the IFAN canal and the period to the resolution of NSD of the IFAN from 3 months to 1 year after SSRO. If the HU values around the IFAN were greater than 300 HU and/or the distance from the buccal aspect of the IFAN canal to the outer buccal cortical margin was less than 6 mm, NSD of the IFAN at 1 year after SSRO was significantly increased. CONCLUSION: It is important to pay particular attention to the anatomical position of the IFAN and the bone quality around the IFAN to predict long-term NSD of the IFAN after SSRO.

Comparison of material-related complications after bilateral sagittal split mandibular setback surgery: biodegradable versus titanium miniplates.

PURPOSE: The aim of the present prospective study was to compare material-related complications using biodegradable and titanium miniplates after bilateral sagittal split mandibular setback surgery. PATIENTS AND METHODS: The subjects included 200 Japanese adults (67 men and 133 women, age range 18 to 45 years) with jaw deformities diagnosed as mandibular prognathism. All patients were prospectively and consecutively randomized to 2 study groups, receiving biodegradable or titanium fixation plates. Of the 200 patients, 110 underwent bilateral sagittal split ramus osteotomy with a biodegradable fixation plate and 90 underwent bilateral sagittal split ramus osteotomy with a titanium metal plate. The clinical records and radiologic findings of the patients were reviewed, and the incidence of material-related complications was compared. RESULTS: The incidence of postoperative complications and breakage in the biodegradable group was 8.2% (9 cases) and in the titanium group was 3.3% (3 cases). No statistically significant difference in the incidence of complications was found between the 2 groups. Fractures of the biodegradable plate occurred at a significantly greater frequency in patients with asymmetry than in patients without asymmetry. CONCLUSION: Biodegradable plates were reliable with minimal material-related complications. However, the use of biodegradable plates should be recommended for minimally loaded situations.

Correlation of mandibular bone quality with neurosensory disturbance after sagittal split ramus osteotomy.

Our aim was to find out whether the quality of bone around the inferior alveolar nerve is correlated with neurosensory disturbance to the nerve after sagittal split ramus osteotomy (SSRO) in patients with mandibular prognathism. Computed tomograms (CT) were taken of 35 patients with mandibular prognathism and 35 without. To assess the density of bone around the inferior alveolar nerve, the width of the buccal cortical bone in the mandibular second molar regions was measured on CT. The Hounsfield units (HU) in the same regions were also measured. The number of HU in the mandible around the second molar regions was significantly higher (p<0.01) in those with neurosensory disturbance (p<0.01). The quality of bone measured by HU is associated with an increased risk of neurosensory disturbance, but the width of buccal bone is not.

NDRG2 is a candidate tumor-suppressor for oral squamous-cell carcinoma.

Oral cancer is one of the most common cancers worldwide, and squamous-cell carcinoma (OSCC) is the most common phenotype of oral cancer. Although patients with OSCC have poor survival rates and a high incidence of metastasis, the molecular mechanisms of OSCC development have not yet been elucidated. This study investigated whether N-myc downstream-regulated gene 2 (NDRG2) contributes to the carcinogenesis of OSCC, as NDRG2 is reported to be a candidate tumor-suppressor gene in a wide variety of cancers. The down-regulation of NDRG2 mRNA, which was dependent on promoter methylation, was seen in the majority of OSCC cases and in several cases of precancerous leukoplakia with dysplasia. Induction of NDRG2 expression in an HSC-3/OSCC cell line significantly inhibited cell proliferation and decreased colony formation ability on soft agar. The majority of OSCC cell lines showed an activation of PI3K/Akt signaling, and enforced expression of NDRG2 in HSC-3 cells decreased the level of phosphorylated Akt at Serine 473 (p-Akt). Immunohistochemical p-Akt staining was detected in 56.5% of the OSCC tumors, and 80.4% of the tumors were negative for NDRG2 staining. Moreover, positive p-Akt staining was inversely correlated with decreased NDRG2 expression in OSCC tumors with moderate to poor differentiation (p<0.005). Therefore, NDRG2 is a candidate tumor-suppressor gene for OSCC development and probably contributes to the tumorigenesis of OSCC partly via the modulation of Akt signaling.

Acceleration of orthodontic tooth movement by alveolar corticotomy in the dog.

INTRODUCTION: Tooth movement and alveolar bone reaction after corticotomies have not been thoroughly examined. In this study, the effects of corticotomies on orthodontic tooth movement and alveolar bone reaction were investigated in dogs. METHODS: Corticotomies were performed on the cortical bone of the mandibular left third premolar region in 12 male adult beagles. The third premolars on the left experimental side and on the right sham side were moved mesially with a continuous force of 0.5 N. RESULTS: Tooth movement velocities from 0 to 1 week and from 1 to 2 weeks after the corticotomies were significantly faster on the experimental side than on the sham side. Hyalinization of the periodontal ligament appeared only at 1 week after the corticotomies on the experimental sides, whereas it was observed from 1 to 4 weeks after the corticotomies on the sham sides. Tartrate-resistant-acid-phosphatase positive cells of the experimental side tended to work vigorously at an early time on the alveolar wall and in the bone marrow cavities. CONCLUSIONS: Orthodontic tooth movement increased for at least 2 weeks after the corticotomies. This might be brought about by rapid alveolar bone reaction in the bone marrow cavities, which leads to less hyalinization of the periodontal ligament on the alveolar wall.

An adult bimaxillary protrusion treated with corticotomy-facilitated orthodontics and titanium miniplates.

We performed an orthodontic treatment combined with corticotomy and the placement of titanium miniplates in an adult patient who desired a shortened treatment period. The patient had an Angle Class I malocclusion with flaring of the maxillary and mandibular incisors. First, titanium miniplates were placed into the buccal alveolar bone of the maxilla for absolute orthodontic anchorage. Second, an edgewise appliance was applied to the maxillary and mandibular teeth. Then, the maxillary first premolars and mandibular second premolars were extracted. At the same time, a corticotomy was performed on the cortical bone of the lingual and buccal sides in the maxillary anterior as well as the mandibular anterior and posterior regions. Leveling was initiated immediately after the corticotomy. The extraction spaces were closed with conventional orthodontic force (approximately 1 N per side). The edgewise appliance was adjusted once every 2 weeks. The total treatment time was 1 year. Cephalometric superimpositions showed no anchorage loss, and panoramic radiographs showed neither significant reduction in the crest bone height nor marked apical root resorption. A corticotomy-facilitated orthodontic treatment with titanium miniplates might shorten an orthodontic treatment period without any anchorage loss or adverse effects.

Reduced syndecan-1 expression is correlated with the histological grade of malignancy at the deep invasive front in oral squamous cell carcinoma.

BACKGROUND: Although many histopathological characteristics of oral squamous cell carcinoma (O-SCC) have been identified as prognostic factors, no factor is completely accurate and unequivocal. This study evaluated the association between the loss of syndecan-1 expression and the histological grade of malignancy at the deep invasive front in O-SCC. METHODS: The expression of syndecan-1 at the invasive tumor front of O-SCC was examined immunohistochemically using archived tissue from 72 cases. The mean age of the patients was 62.5 years (range: 23-90 years) and the male-female ratio was 1.3:1 (41 men, 31 women). There were 26, 24, 11, and 11 cases classified as stages I-IV respectively. The correlation between the intensity of syndecan-1 immunostaining and the clinicopathological factors, especially the histological grade of malignancy at the deep invasive front (invasive front grade) was analyzed. RESULTS: Of the 72 cases, seven (9.7%), 29 (40.3%), 36 (50.0%) showed strong, intermediate, and weak or negative syndecan-1 staining respectively. There were significant differences between syndecan-1 expression and prognosis, differentiation, and pattern of invasion at the deep invasive front. Moreover, the invasive front grade scores, based on the intensity of syndecan-1 staining, were 5.6 +/- 1.0, 8.0 +/- 2.1, and 10.2 +/- 2.3 points with strong, intermediate, and weak or negative intensity respectively; and the difference was significant (P < 0.0001). Patients with intermediate or strong intensity for syndecan-1 had significantly better prognoses than did those with negative or weak intensity (P = 0.0138). CONCLUSION: This study demonstrated that the reduced expression of syndecan-1 seems to be a useful marker of histological malignancy at the deep tumor invasive front and may be a useful prognostic factor in O-SCC.

Relationship between head position and the muscle hardness of the masseter and trapezius muscles: a pilot study.

The purpose of this study was to examine the hardness of the masseter and trapezius muscles at various head positions and to explore the relationship of these two muscles to each other in terms of their respective levels of muscle hardness at different head positions. Twenty-two asymptomatic male subjects participated in this study. Using a hand-held hardness meter, muscle hardness was first measured in a relaxed position as a baseline. The subjects were then asked to assume five deviated head positions, and the muscle hardness was measured again. The data obtained at each deviated head position were compared to those at baseline. In addition, the subjects were asked to maintain a five minute sustained anterior flexion of the head, and muscle hardness was also measured and compared to the baseline. As a result, there was a significant increase in muscle hardness at the point of the whole trapezius with 30-degree anterior flexion, while there was a significant decrease at the point of the right masseter. A significant increase in hardness was seen in the upper trapezius muscle in conjunction with right side bending and in the right upper trapezius muscle on left side bending. Also, a significant decrease in hardness was observed in the right masseter with right side bending. With reference to the axial rotation, there was a significant increase in hardness in the upper right trapezius muscle upon right axial rotation. Moreover, there was a significant increase in the muscle hardness in the whole trapezius after the five minute anteriorly sustained head task. This study provides evidence that deviated head positions lead to an increase in hardness of the trapezius muscle. The data also revealed the simultaneous occurrence of the elevation of muscle hardness in the upper trapezius muscle and the decrease in muscle hardness in the masseter muscle associated with right side bending and anterior flexion.

Analysis of muscle hardness in patients with masticatory myofascial pain.

PURPOSE: The aim of the present study was to access any changes in the muscle hardness of the masseter muscle between normal subjects and patients with myofascial pain during brief sustained isometric contractions at various bite force levels, and to compare muscle hardness, especially in terms of the recovery phase, after a clenching task. MATERIALS AND METHODS: Ten patients with masticatory myofascial pain and 10 age- and weight-matched normal healthy controls participated in this study. First, the hardness of the right masseter muscle was measured at the bite force of 0, 3, 6, and 9 kgf with a hand-held hardness meter. Then, the subjects were requested to exert a 9 kgf-clenching task for 30 seconds. The muscle hardness was again measured at 5, 30, and 120 seconds after the task, and the data obtained were compared with the muscle hardness before the clenching task. RESULTS: The results showed that there was no significant difference between the patients and the normal controls, while the muscle hardness increased with contraction in all subjects. The present findings also showed that the patients had a delayed return to baseline after the clenching task compared with the normal subjects, although an immediate increase after the clenching task was seen in all subjects. CONCLUSION: The results indicated that patients with masticatory myofascial pain have different muscle properties in the recovery phase after contraction, probably because of a slower intramuscular reperfusion.

Effect of short-term use of a centric occlusion stabilization oral appliance on sensory and pain perception thresholds in the cervically innervated area: a pilot study.

This preliminary study was done to determine the effect of short-term use of a centric occlusion stabilization oral appliance with regard to noxious and sensory perception in the upper extremities. The subjects consisted of 22 asymptomatic females, and the experiments were performed on two separate days within the same week, with the days randomly assigned as either appliance-wearing or nonappliance-wearing days. For each experimental day, cool sensation, warm sensation, cold-induced pain and heat-induced pain were measured using a computer-based quantitative testing device, and these thresholds were compared between the experimental days with or without the oral appliance. We found that during the experimental day wearing an oral appliance, subjects had significantly higher thresholds for warm sensation and heat-induced pain. There was no statistical difference between the testing days in cool sensation or cold-induced pain thresholds. These findings indicate that short-term wearing of a centric occlusion stabilization oral appliance may inhibit some noxious and sensory inputs from cervically innervated structures, which are primarily served by unmyelinated C fibers, and that oral appliances may be appropriate for the treatment of painful cervical disorders.