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We describe a case of odontogenic maxillary sinusitis with sulfur granules suspected to be actinomyces. An 86-year-old woman was referred to our hospital for evaluation and treatment of nasal discharge. The clinical diagnosis was odontogenic maxillary sinusitis. We extracted the right maxillary first and second molars. After extaction, the socket was opened to enter the right maxillary sinus.We obtained granulation tissue with sulfur granules from the bottom of the maxillary sinus. Histopathological examination revealed the presence of actinomyces. We irrigated the maxillary sinus with saline from the extraction socket. As of 2 months after the operation, no signs or symptoms have been observed clinically or radiographically.

Epidermal growth factor receptor (EGFR) regulates the growth and progression of human oral squamous cell carcinoma (SCC). Recently, the link between EGFR signaling and the cell cycle has been identified. Some reports have described that EGFR-blocking monoclonal antibody 225 (mAb225) induced G1 arrest and inhibited the growth of various cancer cells. The purpose of this study was to evaluate the effect of mAb225 on human oral SCC cell lines. Exposure to mAb225 in culture inhibited the growth of oral SCC cell lines in an EGFR number-independent manner, with the percent inhibition ranging from 13.8 to 76.6%. Flow-cytometric analysis demonstrated that treatment with mAb225 induced cell accumulation in G1 phase, accompanied by a decrease in the percentage of cells in the S phase. Apoptosis was not seen in this study. G1 arrest was accompanied by a decrease in CDK2-, CDK4-, and CDK6-associated histone H1 kinase activities, and an increase in the expression levels of cell cycle inhibitors p27 KIP1 and p15 INK4B . These results suggested that the antiproliferative effect of EGFR blockade by mAb225 in oral SCC may be mediated by p27 KIP1 and p15 INK4B .

Bio-hybrid artificial organs are an attractive concept to restore organ function through precise biological cooperation with surrounding tissues in vivo. However, in bio-hybrid artificial organs, an artificial organ with fibrous connective tissues, including muscles, tendons and ligaments, has not been developed. Here, we have enveloped with embryonic dental follicle tissue around a HA-coated dental implant, and transplanted into the lower first molar region of a murine tooth-loss model. We successfully developed a novel fibrous connected tooth implant using a HA-coated dental implant and dental follicle stem cells as a bio-hybrid organ. This bio-hybrid implant restored physiological functions, including bone remodelling, regeneration of severe bone-defect and responsiveness to noxious stimuli, through regeneration with periodontal tissues, such as periodontal ligament and cementum. Thus, this study represents the potential for a next-generation bio-hybrid implant for tooth loss as a future bio-hybrid artificial organ replacement therapy.

Lymph node metastasis is a major factor for poor prognosis in oral squamous cell carcinoma (OSCC). However, the molecular mechanisms of lymph node metastasis are unclear. We determined that angiopoietin-like protein 4 (ANGPTL4) mRNA and protein expression were increased in OSCC cells established from the primary site in metastatic cases. In addition, ANGPTL4 expression in biopsy specimens was correlated with the presence of lymph node metastasis. Therefore, our initial findings suggest that OSCC cells expressing ANGPTL4 may possess metastatic ability. Furthermore, cell culture supernatants from OSCC cells that metastasized to the lymph node contain ANGPTL4 and promote invasive ability. These findings suggest that secreted ANGPTL4 may affect the invasive ability of OSCC. Moreover, the rates of positive ANGPTL4 expression at the primary site were significantly higher in the lymph node metastasis group. These results demonstrate that ANGPTL4 contributes to OSCC metastasis by stimulating cell invasion. Therefore, ANGPTL4 is a potential therapeutic target for preventing cancer metastasis.

The junctional epithelium (JE) is an epithelial component that is directly attached to the tooth surface and has a protective function against periodontal diseases. In this study, we determined the origin of the JE using a bioengineered tooth technique. We transplanted the bioengineered tooth germ into the alveolar bone with an epithelial component that expressed green fluorescence protein. The reduced enamel epithelium from the bioengineered tooth fused with the oral epithelium and the JE was apparently formed around the bioengineered tooth 50 days after transplantation. Importantly, the JE exhibited green fluorescence for at least 140 days after transplantation, suggesting that the JE was not replaced by oral epithelium. Therefore, our results demonstrated that the origin of the JE was the odontogenic epithelium and odontogenic epithelium-derived JE was maintained for a relatively long period.

It is important to clarify the participation of periodontal ligament (PDL) cells in the regeneration of alveolar bone to establish a reliable approach for obtaining periodontal regeneration. The aim of this study was to determine whether PDL cells play an important role in alveolar bone repair during the course of periodontal regeneration.In an in vitro study, the expression of the osteoblast phenotype, such as alkaline phosphatase activity and parathyroid hormone-dependent 3',5'-cyclic adenosine monophosphate accumulation, was investigated in dog PDL cells (DPLC) and dog bone cells isolated from mandibles (DBC). In a related study, the roots of mandibular third premolars extracted from aged dogs were divided into a PDL(+) group, in which the PDL was preserved, and a PDL(-) group, in which the PDL was removed. These roots were respectively transplanted into surgically created bone cavities with buccal and interproximal bone defects in an edentulous area, prepared in advance by extraction of mandibular fourth premolars. These bone defects with the transplanted roots were completely covered with submerged physical barrier membranes. New bone formation and new connective tissue attachment, which require new cementum and insertion of functionally oriented new collagen fibers of periodontal ligament, were histomorphometrically assessed, and were compared between the PDL(+) and PDL(-) groups 6 weeks after transplantation.Both cultured DPLC and DBC exhibited the osteoblast phenotype. New connective tissue attachment was observed only in the PDL(+) group. However, alveolar bone was almost completely regenerated to the original bone height in both the PDL(+) and PDL(-) groups, and the amount of newly formed bone was not significantly different between the 2 groups.DPLC retain the capability to differentiate into an osteoblast lineage and may act in the regeneration of periodontal ligament with new cementum formation, whereas these cells may have a limited influence on alveolar bone formation during the course of periodontal regeneration.

癌細胞の悪性度表現として, 1.腫瘍細胞ないし組織の形態の異型性が強いこと。2.腫瘍の増殖が早く, 腫瘍周囲の組織を破壊すること。3.転移を形成することが挙げられる。たとえば, 核クロマチンの増加や不均一性は核細胞質問の物質交換の亢進あるいは異常な相互作用の反映として見られ, その結果, 蛋白合成過程の異常が出現する。核小体の増大は細胞増殖の亢進, より多くの迅速なRNAのターンオーバーの結果としてみられる。同時に, 細胞分裂は細胞増殖性の亢進, 細胞周期の変調, 染色体異常の発生を示唆するものである。この様に細胞が悪性化し, 増殖能を獲得することにより, 細胞には多様な表現が出現する。本研究では, 癌細胞の悪性度表現について, 免疫組織学的および遺伝子解析を行い, 癌細胞の多様な悪性度を検索した。その結果, 悪性度の表現形質は癌細胞の形質をもっとも良く反映している部位である細胞増殖域を検索することが重要であることを示した。

Castleman's disease is a rare, benign lymphoproliferative disorder of unknown origin. The mediastinum is most commonly affected by this disease, followed by the cervical lymph nodes. We report a rare case of Castleman's disease arising in the buccal region. An 81-year-old woman was referred to our hospital in 1989 for further examination and treatment of a slowly-growing left buccal mass. A chicken egg-sized, relatively well-defined painless mass was found in the left buccal region. Smaller stellite masses were also palpable. Computed tomography (CT) revealed a homogenouslyenhanced soft tissue mass in the left buccal region, with expansion to the left infratemporal fossa and pterygomandibular space. The left parotid gland was not involved. A biopsy specimen of the lesion displayed features of Castleman's disease. Immunoserological analyses showed a markedly elevated IgM level. Radiotherapy followed by surgical resection of the lesion was performed. Postoperative follow-up has revealed no signs of recurrence.

Local amyloidosis was found in the right cheek of a 45-year-old woman. The tumor measured 13×18 mm and was located beneath healthy mucosa. It had s hard elasticconsistency and a relatively distinct border. A benign tumor was suspected on the basis of palpation and ultrasonic examination, and the entire tumor was enucleated under local anesthesia. Because the diagnosis of amyloidosis was confirmed histopathologically a comprehensive general physical examination was performed by a physician. Since no other medical abnormalities were found, we confirmed the diagnosis to be local cheek amyloidosis.

舌扁平上皮癌のリンパ節転移を解析するためには, リンパ節高転移能を有する細胞株の樹立が必要である。われわれは, ヒト口腔扁平上皮癌細胞株 (SAS, BSCC) を用いて, ヌードマウス舌への同所移植を繰り返すことにより, リンパ節高転移能を有する口腔扁平上皮癌細胞株の樹立に成功したので, この細胞株の性状解析をあわせて報告する。樹立し得た2種類の高リンパ節転移細胞株ともに, 舌への生着率並びに頸部リンパ節への転移率は100%を示し, もとの細胞株に比べ, 細胞増殖能, 浸潤能, 遊走能ともに亢進がみられ, 特に浸潤能の亢進が顕著であった。免疫蛍光抗体法では, E-カドヘリン, βカテニン, FAKはいずれの細胞株にも弱陽性であったが, 局在の違いは明らかではなかった。Western blotting法による解析では, FAK, パキシリンともに高転移能を有する細胞株において発現量の増加がみられた。リン酸化蛋白質も125kDa付近のものに増加がみられた。以上のことから, 癌細胞のリンパ節転移能の獲得には, 浸潤能の亢進, FAK, パキシリン等のシグナル伝達分子の発現量増加, 蛋白質リン酸化の亢進が重要であることが示された。

Frozen tissue sections of developing and adult rat heads were incubated on a film coated with a gelatin-containing colloidal silver emulsion in order to detect gelatinolytic activity present in the different tissues. The method, termed film in situ zymography, is based on the ability of the thiol group of the propeptide released from the degraded gelatin to induce a structural change in the colloidal silver and thereby a visible change in color. The frozen tissue sections mounted on the coated film were incubated at 37 degrees C overnight. Gelatinolytic activity was detected as a color change from yellow to red. The activity of gelatinase was completely blocked by phenanthroline, which inhibits matrix metalloproteinases. Gelatinolytic activity was widely present in the oral epithelium, tooth buds, tongue, Meckel's cartilage, salivary glands, and other tissues. The intensity of the gelatinolytic activity varied among the different tissue types. The present study demonstrated gelatinolytic activity in both developing and adult craniofacial tissues. These results suggest that gelatinolytic activity plays an important role in normal turn-over in several tissues. Whereas some of the activity also in the developing rats may be related to this turn-over process, some of it is probably directly associated with developmental changes.

Chondrolipoma is a rare form of lipoma with cartilaginous metaplasia. Chondrolipoma of the breast is well known, but only a few chondrolipomas involving the oral region have been reported. To the authors' knowledge, only 3 patients with chondrolipoma involving the lip have been reported in the literature. This report is of a 59-year-old man with chondrolipoma of the lip, who was referred with a painless mass on the left side of the lower lip.

The distribution of fodrin in oral precancerous lesions and squamous cell carcinoma was examined by immunoperoxidase staining. In normal mucous epithelium and leukoplakia, fodrin was localized mainly along the plasma membrane of the granular, prickle and basal cells of the mucous epithelium. Diffused distribution was also observed in the cytoplasm of basal cells. In contrast, staining of fodrin was not clear in well-differentiated squamous cell carcinoma. It decreased markedly at the plasma membrane of the basal-like cells and slightly at the plasma membrane of prickle-like and glanular cells. However, diffused distribution of fodrin appeared in the cytoplasm of prickle-like and granular-like cells. In the cytoplasm of the basal-like cells, labeling of fodrin increased when compared with the basal cells of normal epithelium. In undifferentiated squamous cell carcinoma, fodrin labeling was absent at the plasma membrane of peripheral cells in the cancer nests.The present findings indicate that the pattern of fodrin staining was consistently absent at the peripheral edge of the proliferation area in the early stages of carcinoma and the cancer nest of undifferentiated squamous cell carcinoma, and that fodrin is related to cell differentiation and proliferation.

Salivary gland hypofunction, also known as xerostomia, occurs as a result of radiation therapy for head cancer, Sjögren’s syndrome or aging, and can cause a variety of critical oral health issues, including dental decay, bacterial infection, mastication dysfunction, swallowing dysfunction and reduced quality of life. Here we demonstrate the full functional regeneration of a salivary gland that reproduces the morphogenesis induced by reciprocal epithelial and mesenchymal interactions through the orthotopic transplantation of a bioengineered salivary gland germ as a regenerative organ replacement therapy. The bioengineered germ develops into a mature gland through acinar formations with a myoepithelium and innervation. The bioengineered submandibular gland produces saliva in response to the administration of pilocarpine and gustatory stimulation by citrate, protects against oral bacterial infection and restores normal swallowing in a salivary gland-defective mouse model. This study thus provides a proof-of-concept for bioengineered salivary gland regeneration as a potential treatment of xerostomia., Salivary gland dysfunction as a result of diseases or ageing reduces the quality of life and causes various oral health problems. Here the authors show that the salivary gland function of mice can be recovered by orthotopic transplantation of a bioengineered salivary gland germ.

Introduction Mandibular growth is believed to be strongly related to mastication. Furthermore, mandibular condylar cartilage is known to be derived from neural crest cells. We examined whether the degree of chewing affects condylar cartilage growth of the mandible. Methods Mice were fed diets with varying hardness. Genes specific to neural crest–derived cells were measured by real-time polymerase chain reaction to compare the expression changes between the mandibular and tibia cartilages. The mandibular condylar cartilage was then evaluated histologically, and proliferation was evaluated using proliferating cell nuclear antigen. Immunostaining was conducted for osteopontin, type X collagen, and Musashi1, and real-time polymerase chain reaction was used to assess the expression levels of osteopontin and type X collagen. Results Markers including P75, Wnt-1, Musashi1, and Nestin were upregulated in the mandibular condylar cartilage as compared with the tibial cartilage. Histologic assessment of the mandibular cartilage showed that the hypertrophic chondrocyte zone was statistically significantly thicker in mice fed a hard diet. Chondrocyte proliferation and Musashi1 expression were lower in mice fed a hard diet. After 4 weeks, numerous osteopontin and type X collagen-positive cells were observed in mice fed a mixed diet. Conclusions Mastication affects the balance between differentiation and proliferation in the mandibular condylar cartilage. This phenomenon might be attributed to the presence of neural crest–derived cells.

The MSH6 gene is one of the mismatch repair genes involved in Lynch syndrome and its mutations account for 10-20% of Lynch syndrome. Although previous studies suggested that the difference of the geographical region affects the clinical phenotype of Lynch syndrome, there has been no report on the detailed features of Japanese Lynch syndrome patients carrying an MSH6 mutation. The aim of the present study was to investigate the clinical and molecular features of MSH6 mutation carriers in Japan. Surgically resected 1720 colorectal carcinoma specimens were screened by microsatellite instability (MSI) testing and the MSI-high cases were subjected to a germline mutation analysis of the mismatch repair genes MLH1, MSH2 and MSH6. We investigated the clinical and molecular features of the MSH6 variants, such as the family cancer history, pathological findings, immunohistochemistry, methylation status of the MLH1 promoter and BRAF mutation in the colorectal tumor. Furthermore, the impact of the missense variants on MSH6 protein was predicted by using in silico tools. We identified nine novel pathogenic mutations and eight unclassified missense variants. Among the eight missense variants, three were suspected pathogenic by in silico analysis. We also found that most colorectal cancers in the MSH6 mutation carrier were diagnosed after the age of 50 and were localized distally. Furthermore, the mean age at diagnosis of endometrial cancer in Japanese MSH6 mutation carriers (49.2 years) was earlier than previous reports from Western countries (56.5 years). These results may improve the surveillance program for Japanese MSH6 mutation carriers.

We found that dimethyl-sulfoxide (DMSO) at concentrations of 2.5% induced apoptosis in SV40-immortalized human keratinocytes, while normal keratinocytes were arrested at the boundary of G1/S phase under the same conditions. DMSO-induced apoptosis in SV-40 immortalized keratinocytes was not associated with change in phosphorylated state of the retinoblastoma susceptibility gene. When SV40-immortalized cells were treated with 2.5% DMSO, dissociation of the complex was observed by immunoblotting of SV40 T antigen from immunoprecipitated p53 protein fraction.

Florid cemento-osseous dysplasia is an uncommon disease of cemento-osseous tissue calcification in the tooth-bearing area of the jaw. Although most patients with florid cemento-osseous dysplasia do not have a family history of the disease, a few familial cases have been reported. This report is of a 29-year-old Japanese woman with florid cemento-osseous dysplasia. The patient's wisdom tooth was extracted due to pericoronitis, which was followed by intensive intravenous antibiotic therapy, although surgical procedures are usually not recommended for patients with florid cemento-osseous dysplasia. Radiological examination of the patient's father, who had a history of prolonged osteomyelitis of the upper jaw, also revealed typical features of florid cemento-osseous dysplasia.

Sleep bruxism is a sleep-related movement disorder that can be responsible for various pains and dysfunctions in the orofacial region. The aim of the current case-control association study was to investigate the association of genetic, psychological and behavioral factors with sleep bruxism in a Japanese population. Non-related participants were recruited and divided into either a sleep bruxism group (n = 66) or control group (n = 48) by clinical diagnoses and 3-night masseter electromyographic recordings by means of a portable miniature device. The Epworth Sleepiness Scale, Temperament and Character Inventory, NEO-Five Factor Inventory and custom-made questionnaires that asked about familial aggregation, alcohol intake, caffeine intake, cigarette smoking, past stressful life events, daytime tooth-contacting habit, temporomandibular disorder, daily headache, snoring, apnea/hypopnea symptoms, leg-restlessness symptoms and nocturnal-myoclonus symptoms were administered. In addition, 13 polymorphisms in four genes related to serotonergic neurotransmission (SLC6A4, HTR1A, HTR2A and HTR2C) were genotyped. These factors were compared between case (sleep bruxism) and control groups in order to select potential predictors of sleep-bruxism status. The statistical procedure selected five predictors: Epworth Sleepiness Scale, leg-restlessness symptoms, rs6313 genotypes, rs2770304 genotypes and rs4941573 genotypes. A multivariate stepwise logistic regression analysis between the selected predictors and sleep-bruxism status was then conducted. This analysis revealed that only the C allele carrier of HTR2A single nucleotide polymorphism rs6313 (102CT) was associated significantly with an increased risk of sleep bruxism (odds ratio = 4.250, 95% confidence interval: 1.599-11.297, P = 0.004).This finding suggests a possible genetic contribution to the etiology of sleep bruxism.

The growth and differentiation of human normal keratinocytes and their transformed counterparts were examined in organotypic cultures in which the keratinocytes were grown at the air‐liquid interface on top of contracted collagen gel containing fibroblasts. We developed a modified culture procedure including the use of a mixed medium for keratinocytes and fibroblasts. Normal keratinocytes formed a three‐dimensional structure of epithelium that closely resembled the epidermis in vivo, consisting of basal, spinous, granular and cornified layers. Cells synthesizing DNA were located in the lowest basal layer facing the collagen gel. Expressions of proteins involved in epidermal differentiation were examined by immunohistochemical staining and compared with those in skin in vivo. In the organotypic culture, transglutaminase, involucrin and filaggrin were expressed, as in the epidermis in vitro, most prominently in the granular layer. Type IV collagen, a component of basement membrane, was expressed at the interface between the keratinocyte sheet and the contracted collagen gel. Keratinocytes transformed by simian virus 40 or human papilloma virus (HPV) exhibited a highly disorganized pattern of squamous differentiation. In particular, HPV‐transformed cells invaded the collagen gel. Organotypic culture is unique in that regulatory mechanisms of growth and differentiation of keratinocytes can be investigated under conditions mimicking those in vivo.

The purpose of this study was to histomorphometrically evaluate the use of hydroxylapatite (HA) implants in 60Co-irradiated bone. HA implants were installed in rabbit mandibles 3 months, 6 months, and 12 months after 15-Gy irradiation. Nonirradiated rabbits served as controls. The rabbits were killed 7, 14, 30, 60, and 90 days after the HA implantation. The histologic features of the healing process were examined and histomorphometric measurements were made to quantify the percentage of HA-bone contact and trabecular bone in the medullary cavity. In the irradiated groups, although HA-bone contact was observed later than that in the controls, recovery increased with time after irradiation and the rate of HA-bone contact bone-contacting implant surface ratio; BCSR exceeded 90% in all groups examined before 90 days. In the radiated groups, the average trabecular bone-specific volume was lower than that in the controls and began to decrease before BCSR exceeded 90%. Based on the present data, as well as data from the literature, it is suggested that the success rate of HA implants in irradiated bone increases with the interval after radiotherapy. It is also recommended that HA implants in irradiated bone be installed so that bearing by the cortical bone is increased.

Preeclampsia is still one of the leading causes of maternal and neonatal mortality and morbidity. Despite intensive research, the cause of preeclampsia has not yet been established. However, for a variety of reasons, the numerous aspects of normal and pathological pregnancies, particularly with regard to preeclampsia, remain difficult to study and are, therefore, poorly understood. The development of a laser-based microdissection system has provided rapid morphologically and phenotypically distinct types of cells in the placenta for molecular analysis. Alterations in gene expression in the cytotrophoblast and syncytiotrophoblast or other specific placental cell types from patients who later develop preeclampsia have been reported. Laser microdissection is an attractive method to study each specific placental cell type to characterize the development of preeclampsia or to study the pathophysiology of preeclampsia. This method may contribute to a better understanding of the pathophysiology of preeclampsia.

Objective: Breast cancer cells with CD44+CD24–/low gene expression signature have been suggested to have stem cell-like tumor-initiating properties. The purpose of this study is to clarify the gene expression profiling of cells with CD44+CD24–/low gene expression signature in the luminal subtype. Methods: Laser capture microdissection was used to select the isolation of cancer cells in 35 frozen tissues of breast cancer, and RNA extracted from these cells was examined by real-time RT-PCR to quantify CD44 and CD24 expressions. Human stem cell RT2 Profiler PCR Array was used for gene expression analysis in the groups of CD44+CD24–/low and CD44+CD24+ gene expression signature. Results: Thirty-five tumors were divided into 3 groups. Group A was composed of the CD44+CD24–/low type, in which the ratio of CD44/CD24 was >10.0. Group B was composed of the CD44+CD24+ type, in which the ratio was >0.1 and ≤10.0. In group C, composed of the CD44–/lowCD24+ type, the ratio was Conclusion: This study suggests that the Notch pathway may be an important signaling pathway in luminal subtype with CD44+CD24–/low gene expression signature. In addition, either ALDH1 or SOX2 may be a candidate marker for cancer stem cells in luminal subtype breast cancer.

Interleukin-1β (IL-1β) induces cell death in chondrocytes in a nitric oxide (NO)- and reactive oxygen species (ROS)-dependent manner. In this study, increased production of lactate was observed in IL-1β-treated mouse chondrocytic ATDC5 cells prior to the onset of their death. IL-1β-induced cell death in ATDC5 cells was suppressed by introducing an siRNA for monocarboxylate transporter-1 (MCT-1), a lactate transporter distributed in plasma and mitochondrial inner membranes. Mct-1 knockdown also prevented IL-1β-induced expression of phagocyte-type NADPH oxidase (NOX-2), an enzyme specialized for production of ROS, whereas it did not have an effect on inducible NO synthase. Suppression of IL-1β-induced cell death by Nox-2 siRNA indicated that NOX-2 is involved in cell death. Phosphorylation and degradation of inhibitor of κBα (IκBα) from 5 to 20 min after the addition of IL-1β was not affected by Mct-1 siRNA. In addition, IκBα was slightly decreased after 12 h of incubation with IL-1β, and the decrease was prominent after 36 h, whereas activation of p65/RelA was observed from 12 to 48 h after exposure to IL-1β. These changes were not seen in Mct-1-silenced cells. Forced expression of IκBα super repressor as well as treatment with the IκB kinase inhibitor BAY 11-7082 suppressed NOX-2 expression. Furthermore, Mct-1 siRNA lowered the level of ROS generated after 15-h exposure to IL-1β, whereas a ROS scavenger, N-acetylcysteine, suppressed both late phase degradation of IκBα and Nox-2 expression. These results suggest that MCT-1 contributes to NOX-2 expression via late phase activation of NF-κB in a ROS-dependent manner in ATDC5 cells exposed to IL-1β.

Preeclampsia is still one of the leading causes of maternal and neonatal mortality and morbidity. Despite intensive research, the cause of preeclampsia has not yet been established. However, for a variety of reasons, the numerous aspects of normal and pathological pregnancies, particularly with regard to preeclampsia, remain difficult to study and are, therefore, poorly understood. The development of a laser-based microdissection system has provided rapid morphologically and phenotypically distinct types of cells in the placenta for molecular analysis. Alterations in gene expression in the cytotrophoblast and syncytiotrophoblast or other specific placental cell types from patients who later develop preeclampsia have been reported. Laser microdissection is an attractive method to study each specific placental cell type to characterize the development of preeclampsia or to study the pathophysiology of preeclampsia. This method may contribute to a better understanding of the pathophysiology of preeclampsia.

Although necrotizing sialometaplasia occurs mostly in the palate, we recently treated a case in which this condition had developed in the distomolar glands of the lower jaw. Only one other similar case has been reported. The present case was apparently caused by chronic, mechanical stimulation by a maxillary molar and denture. Because necrotizing sialometaplasia resolves spontaneously, no special treatment is necessary. However, to diagnose the present case, we totally resected the lesion. The patient was cured, and there have been no signs of recurrence.

In the clinical field of jawbone formation, the use of autogenous bone as the graft material is the gold standard. However, there are some problems with this technique, such as risk of infection on the donor side, the limited amount of available bone mass, and marked resorption of the grafted bone. We investigated the potential for using teeth as a bone graft material for jawbone formation because the dental pulp contains stem cells, including undifferentiated neural crest-derived cells.Alveolar bone defects were created in Wistar rats, and the defects were filled with either tooth or iliac bone graft material, or left as controls. The potential for using teeth as a bone graft material for jawbone formation was measured using real-time polymerase chain reaction, microcomputed tomography, and histologic analysis.Polymerase chain reaction revealed that the expressions of P75, P0, nestin, and musashi-1 were significantly higher in teeth than in mandibular bone and iliac bone grafts. Hematoxylin and eosin staining and microcomputed tomography showed that at 8 weeks, tooth graft material produced a similar amount of new bone compared to iliac bone graft material. Osteopontin was expressed in both the tooth and iliac bone graft material at 6 and 8 weeks after surgery. Dentin sialoprotein was expressed in the tooth graft material in the new bone at 6 weeks only.These results indicate that teeth may be an alternative material to autogenous bone for treating alveolar bone defects by grafting.

To examine early changes in the cell membrane caused by treatment with 9.10-dimethyl-1.2-benzanthracene (DMBA), rat lingual epithelium in vitro was observed by means of electrophysiolo-gical, freeze-fracture and whole mount electron microscopic methods. Membrane potential showed a transient decrease at 6 h after the treatment but returned to almost the same level as that of the control group 1 day later. Membrane resistance decreased rapidly at 6 h after the treatment but increased at 12 h. Transient decrease of membrane resistance and membrane potential occurred prior to the decrease in coupling ratio at 6 h after treatment with DMBA. In freeze-fracture replicas, after 6 h of treatment with DMBA, the number of intramembranous particles (IMPs) was less than that of the control. This decrease was especially pronounced on the P-face. After 12 h treatment with DMBA, the number of IMPs increased by 20-40 % more than the control value. This increase was especially noticeable on the E-face, which was 70 % of the control value. At 3 days and thereafter, the number of IMPs decreased again and remained approximately the same as that of the control, though the Eface values remained larger than those of the control. Treatment with DMBA had a marked effect on the cytoskeletal organization. Treatment with DMBA caused morphologic changes as early as 6 h. These early changes might represent triggers for subsequent metabolic cellular abnormalities. The affected cells which were treated with DMBA appear to differ from normal cells through accelerated-abnormal cell membrane and cytoskeletal turnover.

S-1, an oral fluorouracil antitumor drug, is composed of three agents: tegafur (FT), 5-chloro-2,4-dihydroxypyridine (CDHP), and potassium oxonate (Oxo). Approximately 50% of oral squamous cell carcinomas (OSCC) exhibit cervical lymph node metastasis. The extent of lymph node involvement is a major determinant in both staging and prognosis of the majority of OSCC. The purpose of this study was to examine the effect of S-1 on the metastatic potential of OSCC cells. We used orthotopic green fluorescence protein (GFP) SAS-L1, in BALB/c nu/nu mice. Mice received oral doses of either 5% hydroxypropylmethylcellulose (HPMC) for control or S-1 (20 mg/kg) and were autopsied at 2 weeks. We also performed in vitro experiments using concomitant 5-fluorouracil (5-FU) and CDHP as a drug model of S-1 to determine the effect of S-1 on OSCC invasion and metastasis. Although 100% (11 of 11) of mice not treated with S-1 showed cervical lymph node metastasis, only 54.4% (6 of 11) of S-1 treated mice demonstrated metastasis. In in vitro experiments, OSCC cells treated with 5-FU and CDHP showed a marked reduction in invasiveness and in adhesion to laminin coated plates. Western blot analysis revealed that treatment with 5-FU and CDHP suppressed expression of integrins alphav, alpha3, alpha6, beta1, beta3, beta4, beta5, and beta6. These results suggest that S-1 inhibits tumor proliferation and lymph node metastasis in OSCC cells. Moreover, expression of integrin subunits and the integrin signal transduction pathway may be closely related to metastasis suppression.

It is well known that mastication has a significant influence on mandibular growth and development, but the mechanism behind this effect has not yet been clarified. Furthermore, no studies have examined the effects of changes in mastication on the three-dimensional (3D) morphometry of the mandible. The aim of the present study was to investigate the influences of changes in mastication on mandibular growth and morphology. Twenty-five 3-week-old (at the time of weaning) imprinting control region mice were randomly divided into three groups: mice fed a hard diet (HD), mice fed a soft diet (SD), and mice alternately fed hard and soft diets (HSDs) every week for 4 weeks. The morphometry of the mandible was analysed using 3D microcomputed tomography (μCT). Statistical analysis was undertaken using a t -test. μCT analysis showed that the condylar width was significantly greater in the HD group than in the SD group after 1 week. After 4 weeks, mandibular length was significantly longer and ramus height was greater in the HSD group than in the other two groups. Bone volume was significantly less in the SD group than in the other two groups after 4 weeks. These findings suggest that changes in mastication markedly affect mandibular condylar cartilage growth and mandibular morphology. It is considered that dietary education at an early age is important in order to prevent disruption of the development of the mandible.

Objective Calpain is a calcium ion-dependent cysteine protease, consisting of two primary isoforms (calpain1/calpain2) which mediate crucial cellular functions. The activity of the calpains is tightly regulated by the endogenous inhibitor calpastatin. Calpains have been detected in several studies during the embryonic and foetal stages. The aim of this study is to investigate the temporal transition of typical calpains and their inhibitor calpastatin during odontogenesis. Design We used the first molar of foetal ICR mice from embryonic day (E) 14 to postnatal day (PN) 7. Using laser microdissection and semi-quantitative real-time PCR, we investigated calpain1, calpain2 and calpastatin expressions in each enamel epithelium, inner enamel epithelium, stellate reticulum and outer enamel epithelium. Results We found calpain1 and calpain2 mRNA increased in the all enamel epithelia between E18 and PN1. In addition calpastatin mRNA expression increased in the ameloblasts from PN1 to PN7. The immunohistochemistry results demonstrated that calpain1/calpain2 was present in the distal side of ameloblasts from PN1 to PN7, and calpastatin was present in the extracellular enamel matrix from E16 to PN1. Furthermore calpain1/calpain2 was present in the dentin, and calpastatin was detected in dentin producing odontoblasts and predentin at PN7. Conclusions In this study the temporal transition of calpain1, calpain2 and calpastatin mRNA and the immunolocalization are identified during tooth development. Our results indicate that the calcium-dependent proteases may play an important role in mouse molar development and extracellular calpain and calpastatin may be involved in molar mineralization.

Although RAD21 is involved in the repair of DNA double-strand breaks and is essential for mitotic growth, its role in cancer has been unclear. Recently, cleaved products of RAD21 by caspase 3 and 7 become a factor inducing apoptosis. And there are some reports about the association of tumor formation and RAD21. Our current study indicates the RAD21 gene was closely related to the invasion and metastasis of the cancer cells. In addition, although it is known that there are many cleavage products, about those functions, it is not clear. In this study, the relevance of RAD21 cleaved products expression to cell differentiation and carcinogenesis of oral squamous epithelium was clarified using Laser Microdissection and Western Blot analysis.

Infiltrating pattern is one of important histopathological prognostic factor in head and neck squamous carcinomas. In this study we have assessed differentially expressed genes in each infiltrating patterns by laser microdissection and cDNA microarray. We classified infiltrating patterns of oral squamous cell carcinoma into major three groups; INFα, INFs, and INFγ, based on Japanese General Rule for Clinical and Pathological Studies on Cancer, and collected cancer cells of each infiltrating pattern by laser microdissection. Total RNA was extracted from collected cells and was reverse-transcribed. PCR-based amplification of cDNA, cDNA probe labeling and array hybridization were conducted. We compared expression profiles of cancer cells of each infiltrating pattern to that of normal mucosal epithelium using cDNA microarray consisting of 8327 human genes. We identified several genes that were commonly up- or downregulated in each infiltrating pattern. The differentially expressed genes were growth-related proteins, oncogenes, enzymes and nuclear proteins. The list of genes identified in these analyses provides potentially valuable information for preoperative assessment of evaluation of malignancy and represents a source of novel targets for cancer therapy.

Background Neovascularization in the carotid atherosclerotic plaque is a common pathogenetic feature in carotid artery stenosis. To investigate whether the neovascular region of the stable plaque differentially expresses specific genes, we analyzed the patterns of angiogenesis-related gene expression in regions of the plaque isolated by laser microdissection and examined by immunohistochemistry and real-time reverse transcription (RT)–polymerase chain reaction (PCR). Methods Carotid plaque samples were obtained by carotid endarterectomy in 27 clinically asymptomatic patients with high-grade internal carotid artery stenosis. Among these 27 plaque samples, 23 plaques were confirmed to be stable pathologically, and 14 stable plaques had neovascularization. The medial, shoulder, and neovascular regions of the 14 carotid plaques were determined by immunohistochemical staining. These 3 regions were microdissected, and total RNA was extracted for real-time RT-PCR analysis. The expressions of hypoxia inducible factor 1α, vascular endothelial growth factor-A, thioredoxin, and thioredoxin interacting protein were analyzed at mRNA level. Conclusions Real-time RT-PCR was performed on 42 laser microdissected regions of 14 plaques. The expressions of all four genes examined were significantly lower in the medial region at mRNA level. High expressions were noted in both shoulder and neovascular regions, with no significant difference between the two. Furthermore, these expression patterns were related significantly to macrophage infiltration. In conclusion, hypoxia- and thioredoxin-related genes are significantly overexpressed in human stable carotid atherosclerotic plaques and strongly correlate with macrophage infiltration rather than neovascularization. Macrophage infiltration may lead to overexpression of these genes and promote angiogenesis in stable carotid plaques.

Mandibular condylar cartilage can be distinguished from articular and growth cartilages of long bones based on several differences in morphology, physiology, and function between these structures. However, there is almost no information available on the types of genes that contribute to these differences. In this study, genes that were differentially expressed in mandibular condylar and growth cartilages in 1-week-old rats were investigated using fluorescent differential display (FDD) and laser microdissection (LMD). A number of genes were identified by FDD including chondromodulin-1 (ChM-1), which is known to be an angiogenesis inhibitor of endochondral ossification. ChM-1 expression was then compared with that of tenomodulin (TeM) in mandibular condylar and tibial cartilages of 1- and 5-week-old rats using real time PCR (RT-PCR), immunohistochemistry, and in situ hybridization. There was negligible detection of ChM-1 mRNA and protein in mandibular condylar cartilages compared to tibial cartilages of 1- and 5-week-old rats. On the other hand, TeM mRNA was more abundant in mandibular condylar cartilage than in tibial. These observations demonstrated that gene expression in mandibular condylar cartilage differed from other types of cartilage such as articular and growth ones.

Focal adhesion kinase (FAK), a protein tyrosine kinase, has recently been suggested to play a role in signal transduction through integrins. In fact, FAK is involved in cell proliferation and cell motility by performing signal transduction through integrins. FAK-related non-kinase (FRNK) has been found to be an inhibitor of FAK. As the expression level of FRNK in the cell is very low, the study of FRNK has been preferentially performed by gene overexpression, up to the present, and the role of constitutive FRNK in cells remains unclear. We hypothesized that FRNK is involved in the adhesion of cells to the extracellular matrix (ECM) and investigated the expression of FRNK by time kinetic analysis shortly after cell seeding. We found that FRNK expression was significantly increased in the cells during the early stage of cell adhesion to the ECM. These data indicated that FRNK plays an important role in cell adhesion during the very early stages of cell culture.

A rare case of mucous retention cyst in the middle of the oral floor is reported. A 63-year-old woman was referred because of a painless swelling in the middle of the oral floor, which had persisted for 1 month. Resection was performed. The mass was located between the genioglossus muscle and geniohyoid muscle and was unrelated to the sublingual gland or Warthon's duct.The histopathological diagnosis was solitary mucous retention cyst. Differential diagnosis is also discussed.

This study reports a discharging method for bone-like carbonated HA (cHA)-coating (Ca/P 1.71) and stoichiometric HA (sHA)-coating (Ca/P 1.67) with micrometer order thicknesses on titanium plates, using modified body fluid and acidic calcium phosphate solutions, respectively. In vivo histological performance of the HA coatings prepared by discharging in electrolytes was evaluated. Bone-contact indexes of HA coatings were measured microscopically. Additionally, bone-coating interface was analyzed by scanning electron microscopy and the use of an electron probe microanalyzer. Results demonstrated that there was no significant difference in contact index between HA coatings. However, the cHA coating was practically replaced by immature bone, and the titanium metal substrate was directly connected to the bone structure whereas the sHA coating layer remained and was partially detached from the titanium metal substrate. Since detached coating particles are pathogens, and can cause peri-implantitis, the cHA coating was more favorable than the sHA coating even if contact index was equivalent to that of the sHA coating. It is thought that coating thickness and chemical composition of coatings are important for biological stability of implants. In conclusion, since bone-like thin cHA coating showed high osteoconductivity and bone replacement, bone-like HA is superior to sHA coating for use in dental implants.