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Porphyromonas gingivalis (P. gingivalis) is a Gram-negative anaerobe involved in the pathogenesis of chronic periodontitis, including local inflammation of the oral cavity. However, periodontal disease has recently been identified as a significant factor in the pathogenesis of neural diseases, including Alzheimer's disease. A virulence factor, P. gingivalis-lipopolysaccharide (LPS-PG), is involved in pro-inflammatory responses, not only in peripheral tissues but also in the brain. In this study, we examined whether P. gingivalis-induced brain inflammation could be ameliorated by pharmacotherapy, using in vivo and in vitro studies. In an animal experiment, peripheral administration of LPS-PG induced inflammation in the hippocampus via microglial activation, which was inhibited by pre-treatment with the antidepressant imipramine. Similarly, LPS-PG-induced inflammation in MG-6 cells, a mouse microglial cell line, was inhibited by pre-treatment with imipramine, which caused imipramine-induced inhibition of NF-κB signaling. Culture media obtained from LPS-PG-treated MG-6 cells induced neuronal cell death in Neuro-2A cells, a mouse neuroblastoma cell line, which was prevented by pre-treatment of MG-6 cells with imipramine. These results indicate that imipramine inhibits LPS-PG-induced inflammatory responses in microglia and ameliorates periodontal disease-related neural damage.

The diagnosis of ameloblastic carcinoma is based on a combination of cytological features of malignancy and histological pattern of an ameloblastoma. We report a very rare case of ameloblastic carcinoma arising from a pre-existing ameloblastoma of the maxilla with challenging diagnostic aspects due to atypical histology. A 74-year-old female with a swelling in the right maxillary region was referred to our hospital in March 2015. Intraoral examination revealed a 45 × 30 mm sized swelling extending from the right upper first premolar to the maxillary tuberosity. Radiographic examination revealed a tumor mass with heterogeneous enhancement from the right maxillary alveolar area to the maxillary sinus, invading the pterygopalatine fossa under the cranial base, with bone destruction of the alveolar bone, floor of the maxillary sinus, and pterygoid plate. Incisional biopsy results revealed round or short spindle cells with clear cytoplasm, with low mitotic activity, cellular atypism, and Ki-67 labeling index. A low grade malignant epithelial tumor was suspected. Tumor resection with hemimaxillectomy and partial mandibulectomy was performed in April 2015. The surgical specimen was composed of diffusely distributed spindle cells and clusters of round cells with clear cytoplasm, similar to the biopsy findings. Although cellular atypism was scant, Ki-67 labeling index was high in most sections. Ameloblastoma was detected in part of the tumor and a definitive diagnosis of ameloblastic carcinoma was made. The patient died of metastatic colon cancer 3 years and 10 months post-operatively; there was no evidence of recurrence and metastasis of ameloblastic carcinoma in that duration.

Objectives Lactoferrin (LF) possesses diverse biological functions. We previously reported that bovine LF (bLF) attenuates lipopolysaccharide-induced bone resorption in osteoblasts. In addition to its ability to inhibit osteoclastogenesis, bLF has been implicated in stimulating bone formation. However, the molecular mechanisms of bLF in bone cell anabolism remain unclear. Here, we tried to analyse the molecular mechanisms involved in osteogenesis in the presence of bLF. Methods Alkaline phosphatase activity, Runx2 activity, gene expression, and Alizarin red staining were analyzed to evaluate the osteogenic differentiation status. The expression of the Smads and mitogen-activated protein kinase (MAPK) signaling molecules was analyzed via western blotting. Ex vivo organ cultures of mouse calvariae were performed to evaluate the effect of bLF on bone regeneration. Results bLF enhanced the osteoblastic differentiation of mesenchymal stem cells through activation of Smad2/3 and p38 MAPK, which increased the transcriptional activity of Runx2. bLF treatment also enhanced osteoblastic differentiation and mineralized nodule formation of osteoblast-lineage cells, and repaired bone defects ex vivo. Moreover, inhibition of Smad2/3 or p38 MAPK signaling reduced the anabolic effects of bLF. Together, these results suggested that bLF is a potent osteogenic factor, which mediates its function via activation of the Smad2/3 and p38 MAPK signaling pathways. Conclusions Here, we described a novel function of bLF and its signal transduction mechanisms in osseous tissue. Along with inhibiting osteoclastogenesis, bLF may limit further osteoclast formation and contribute to bone mass enlargement. Thus, bLF represents a potentially valuable therapeutic agent for bone regeneration and destructive bone diseases.

Epithelial-to-mesenchymal transition (EMT) is a biological process of invasion and metastasis in cancers, including in oral squamous cell carcinoma (OSCC). However, an effective anticancer drug that directly targets EMT has not yet been discovered. Therefore, we aimed to investigate the repressive effects of bovine lactoferrin (bLF) on EMT to achieve mesenchymal-to-epithelial transition (MET) in OSCC. OSCC cell lines, HOC313 (EMT-induced) and SCCVII (without EMT induction), were treated with bLF. The effects of bLF on EMT in OSCC were identified histologically by haematoxylin and eosin staining and observed morphologically and immunohistochemically using an anti-E-cadherin antibody. Expression levels of E-cadherin and vimentin were investigated using RT-PCR and western blotting. Immuno-expression of E-cadherin was examined in vivo in tumour tissues of C3H/HeN mice, transplanted with SCCVII cells, with or without bLF administration. We found that bLF changed the spindle-like mesenchymal cells to cuboidal-like epithelial cells and enhanced the affinity of membrane-bound E-cadherin in HOC313 cells. The transformation of EMT-MET in HOC313 cells was confirmed by the upregulation of E-cadherin and suppression of vimentin. Moreover, bLF suppressed TWIST expression through downregulation of ERK1/2 phosphorylation. Additionally, the inhibition tumour cell infiltration and increase in E-cadherin expression were observed in xenografts of the mice orally administered with bLF. Thus, based on the results from in vitro and in vivo studies, we concluded that bLF caused the restoration of epithelial properties through MET. Importantly, this finding is novel and is the first report indicating that bLF inhibited EMT and induced MET in OSCC, suggesting that bLF may provide a novel therapeutic strategy in OSCC.

Bisphosphonates (BP) are potent inhibitors of the osteoclast activity used in the treatment of metastatic bone disease and osteoporosis. Bisphosphonate related osteonecrosis of the jaw (BRONJ) is a rare, but severe complication as a pathological fracture, pain and tumor-induced hypercalcemia. Patients taking BP may subsequently develop BRONJ after dentoalveolar surgery or trauma of the oral mucosa. However, appropriate approaches for the prevention and treatment of BRONJ have not been established. One of reasons is that the pathogenesis of BRONJ is poorly understood. We reported a case of BRONJ in a 77-year-old woman that was successfully treated by segmental resection of the extensive necrotic bone and a pathological fracture in the mandible. We describe the rare histopathological features of the resected specimen.

Objectives Liposarcoma(LS) is the most common soft-tissue sarcoma. The histological spectrum has a well-differentiated-liposarcoma(WDLS) and a more aggressive dedifferentiated-liposarcoma(DDLS). Advanced therapeutic strategies based on molecular mechanism are urgently needed, especially for DDLS. Previously, we reported that TIMP-1 (a member of tissue-inhibitor-of-metalloproteinase), with its receptor CD63 activates yes-associated protein (YAP) and transcriptional co-activator with PDZ binding motif (TAZ) to promote cancer cell proliferation. Aberrant YAP/TAZ activation in LS is reported, however, contribution of TIMP-1-YAP/TAZ axis in LS remains unclear. Intriguingly, TIMP-4 is known to share CD63 as TIMP-1, but its role in LS is unknown. Here we clarified the expression and function of TIMP-1 and -4 through YAP/TAZ regulation in LS. Materials & Methods Cell lines of WDLS(94T778) and DDLS(SW872) were used for in vitro experiments such as Western blotting, RT-PCR, cell-proliferation, migration and apoptosis assay. Results Database analysis showed high TIMP-1 expression in DDLS patients correlating with poor prognosis, while high TIMP-4 expression in WDLS patients with better prognosis. TIMP-1 knockdown in DDLS cells inactivated YAP/TAZ and suppressed cell-growth, migration, which was rescued by constitutively active form of YAP5SA. On the other hand, cell-growth and migration were significantly increased in TIMP-1 over expressing WDLS cells, which was suppressed by verteporfin (a YAP/TAZ inhibitor). TIMP-4 knockdown in WDLS activated YAP/TAZ, promoted cell-proliferation and migration, which was inhibited by verteporfin treatment or YAP/TAZ knockdown. Recombinant TIMP-4 showed opposite results in DDLS cells significantly.TIMP-4 CD63 binding inactivated YAP/TAZ in WDLS. Conclusion The switching of TIMP-4 to -1 expression during transition from a WDLS to a DDLS led to activation of YAP/TAZ and promoted cell-proliferation, migration, inducing poor prognosis. TIMP-1 and -4 as novel YAP/TAZ regulators may warrant future possibilities of targeting key molecules in development of diagnostic and therapeutic novelties in treating LS.

Objective The aim of the study was to clarify the mechanisms underlying orthodontically induced root resorption by characterizing the role of PGE 2 induced by compressive stress (CS) and tensile stress (TS) on cementoblast metabolism in vitro . Design Mouse cementoblast cell line OCCM-30 was continuously stimulated with 0.2 KPa CS or 5.0 KPa TS. COX-2 mRNA expression and PGE 2 production were thus quantified. In addition, cells were treated with COX-2 inhibitor and the role of PGE 2 induced by CS or TS on the expression of genes related to cementoblast differentiation was examined. PGE 2 receptors mRNA expression induced by CS or TS was also evaluated. Moreover, cells were treated with exogenous PGE 2 and the role of PGE 2 concentration on matrix mineralization was verified. Results CS and TS enhanced COX-2 mRNA expression and PGE 2 production. PGE 2 synthesis, however, was markedly induced by CS. Gene expression of bone morphogenetic protein 2 (BMP-2), osteocalcin (OCN) and receptor activator for nuclear factor kappaB ligand (RANKL) was enhanced by CS on an endogenous PGE 2 -mediated manner. Osteoprotegerin (OPG) expression was not affected by CS. Meanwhile, TS up-regulated the expression of BMP-2 and alkaline phosphatase (ALP) on an endogenous PGE 2 -mediated manner. TS down-regulated RANKL mRNA expression, whilst OPG expression was not affected. Moreover, EP4 mRNA expression was considerably enhanced by TS. Regarding PGE 2 concentration, only cells treated with low concentration presented anabolic response. Conclusions Gene expression was differentially regulated according to the type of mechanical stimulation applied to cementoblasts. In addition, it is shown that PGE 2 plays an important role on mediating cementoblast mechanosensitivity.

Objective This study investigates the effect of the N-terminal region of a synthetic porcine ameloblastin peptide on the proliferation and differentiation of human periodontal ligament cells (PDLC). Design We used a cell counter to assess the effect of ameloblastin peptides on the proliferation of PDLC. To investigate the effect of ameloblastin peptides on the differentiation of PDLC, we examined quantitative analysis of alkaline phosphatase (ALP) activity by the Bessey–Lowry enzymological method, mineral nodule formation by Dahl's method, and expression of mineralization-related genes by RT-PCR. We used an anti-ameloblastin antibody to determine whether stimulation of ALP activity was caused by the peptide. Results At all concentrations examined, the effect of the ameloblastin peptide on cell proliferation was not significantly different compared with the control. However, the peptide significantly stimulated ALP activity in a dose-dependent manner. ALP activity was significantly inhibited by an anti-ameloblastin antibody, which caused ALP levels to revert to their approximate levels in the untreated condition. At concentrations greater than 1 ng/ml, the peptide promoted mineralized nodule formation of PDLC. And the peptide induced higher expressions of ALP and bone sialoprotein (BSP) than the control. Conclusion Our results show that the ameloblastin peptide upregulate ALP and BSP levels and can enhance calcification of PDLC. Thus, we suggest that the N-terminal synthetic ameloblastin peptide promotes the differentiation activity of PDLC.

Objective To immortalize human dental pulp (HDP) cell showing stable growth and high mineralization activities in vitro . Design HDP cells were obtained from a healthy third molar and immortalized by transfection with human telomerase transcriptase (hTERT) gene. To examine the characters of hTERT transfected HDP (HDP-hTERT) cells, we examined expression of mRNA for dentin sialophosphoprotein (DSSP), type I collagen (COLI), alkaline phosphatase (ALP) and bone sialoprotein (BSP) by RT-PCR. In addition, we examined ALP activity by biochemical method and nodule formation by alizarin red S (ALZ) staining. Results HDP-hTERT was obtained by transfection with hTERT gene. These cells bypassed the senescence and grew over 120 population doublings (PDLs) without significant growth retardation. High expression of hTERT was confirmed in HDP-hTERT by RT-PCR and showed remarkable telomerase activity. Both HDP-original (HDP-ori) and HDP-hTERT expressed DSSP, COLI, ALP and BSP mRNA and showed ALP activity and ALZ staining at the same levels. Conclusions We were able to establish a cell line of immortalized human dental pulp cells with odontoblastic differentiation which will be a useful cell model for studying the mechanism of proliferation and differentiation of odontoblasts.

Defining the regulatory mechanisms promoting differentiation and proliferation of cementoblasts has not been well understood, because of the lack of cell models in vitro. To establish an in vitro cell model for the cementoblasts, extracted rat molars obtained from 8-week-old rats were used. Cells lining the root surface (cemetoblasts) were obtained by an enzymatic digestion method, and immediately immortalized by transfection of thermolabile SV40 T-antigen gene. The transfected cementum lining cell clones, RCM-C3 and -C4, were maintained for more than 200 population doublings (PD), while the original cells stopped their growth at 60 PD. Thus, immortalized cell lines decreased expression of SV40 T-antigen and subsequently cell proliferation at non-permissive temperature (39 degrees C). Reverse-transcribed-polymerase chain reaction indicated expression of gene for type I collagen, alkaline phosphatase (ALP), osteopontin, and osteocalcin mRNA at both permissive (33 degrees C) and non-permissive (39 degrees C) temperatures. RCM-C4 expressed higher bone siaploprotein (BSP) mRNA than RCM-C3, and further RCM-C4 showed higher BSP mRNA at 39 degrees C than 33 degrees C. High ALP activity and mineralized nodule formation were observed at 39 degrees C in both cell lines. These findings suggested that the cell lines, RCM-C3 and -C4, are useful model for studying the regulatory mechanisms of differentiation and proliferation of cementoblasts.

Oral squamous cell carcinoma (OSCC) is the most frequent malignant neoplasm of the head and neck region. Conversion of normal cells to cancer cells is achieved through a multi-step process that is closely associated with the accumulation of multiple gene changes including both oncogenes and tumour suppressor genes. The proliferation and progression of cancer may be caused by abnormalities of various positive and negative cell cycle regulators. Cell cycle progression is positively regulated by multiple cyclins and cyclin-dependent kinases (Cdks) and cyclin/Cdk complexes are negatively regulated by a number of Cdk inhibitors including p27. p27 is a Cdk inhibitor and plays an important role in negative regulation of the cell cycle during G0 and G1 phases. Degradation of p27 is a critical event for the G1/S transition and occurs through ubiquitination by SCF(Skp2) and subsequent degradation by the 26S proteasome. It has been revealed that down-regulation of p27 is frequently found in various cancers, including OSCC, and is due to an enhancement of its degradation. Importantly, down-regulation of p27 is well associated with its malignancy including poor prognosis in various cancers. Moreover, aggressive human cancers express low levels of p27 because of its decreased stability. More recent evidence suggests that Skp2 and Cks1, the specific recognition factors for p27 ubiquitination, have oncogenic properties. This review will focus on down-regulation of p27 and mechanism of its down-regulation in OSCC.

Down-regulation of p27 is frequently observed in various cancers due to an enhancement of its degradation. Skp2 is required for the ubiquitination and consequent degradation of p27 protein. Another protein called Cks1 is also required for p27 ubiquitination in the SCFSkp2 ubiquitinating machinery. In the present study, we examined Cks1 expression and its correlation with p27 in oral squamous cell carcinoma (OSCC) derived from tongue and gingiva. By immunohistochemical analysis, high expression of Cks1 was present in 62% of OSCCs in comparison with 0% of normal mucosae. In addition, 65% of samples with low p27 expression displayed high Cks1 levels. Finally, Cks1 expression was well correlated with Skp2 expression and poor prognosis. To study the role of Cks1 overexpression in p27 down-regulation, we transfected Cks1 with or without Skp2 into OSCC cells. Cks1 transfection could not induce a p27 down-regulation by itself, but both Cks1 and Skp2 transfection strongly induced. Moreover, we inhibited Cks1 expression by small interference RNA (siRNA) in OSCC. Cks1 siRNA transfection induced p27 accumulation and inhibited the growth of OSCC cells. These findings suggest that Cks1 overexpression may play an important role for OSCC development through Skp2-mediated p27 degradation, and that Cks1 siRNA can be a novel modality of gene therapy.

p27, a cyclin-dependent kinase inhibitor, plays an important role in the negative regulation of the cell cycle during G1-S phases. In our laboratory, we examined expressions of p27 and related factors in oral squamous cell carcinoma (OSCC) and adenoid cystic carcinoma (ACC) to discuss the diagnostic and therapeutic significance of p27 and its related factors in oral malignancies. Our studies demonstrated the following : 1. Reduced expression of p27 is observed in 87% of OSCCs and 84% of ACCs. 2. There is a strong correlation between reduced expression of p27 and poor prognosis of patients with OSCC and ACC. 3. Reduced expression of p27 in malignancies may be caused by post-translational ubiquitin-mediated degradation. 4. High expression of Skp2, an F-box protein specific to p27, is correlated with poor prognosis in OSCC and an inverse correlation between the expression of Skp2 and p27 was observed. 5. Proteasome inhibitors induce apoptosis of OSCC cells through p27 accumulation. 6. Transfection of wild and degradation-resistant mutant types of the p 27 gene inhibits the growth of OSCC cells. These results indicate that p27 and its related factors play important roles in the development of OSCC and can be novel therapeutic targets for oral malignancies as well as strong prognostic markers.

The aim of the present study has been to determine the role of CD44v9 in the metastatic process of oral squamous cell carcinoma (OSCC). We have examined the expression intensity of CD44v9 in four OSCC cell lines, and using cell culture insert investigated the invasion ability of the cells expressing CD44v9 at higher levels (HSC-2, HSC-3), and the cells expressing this protein at lower levels (HSC-4, KB) with or without the treatment with an anti-CD44v9 antibody. In the highly expressing cells, the addition of anti-CD44v9 antibody enhanced their invasion ability, whereas it showed no effect on the invasion ability of the weakly expressing cells. These results suggest that the reduction of CD44v9 expression may weaken cell-to-cell adhesion in OSCC and make the tumor cells detach easily from their nests, resulting in the enhancement of their invasion ability. It may ultimately promote the establishment of a metastatic lesion.

Human cementifying fibroma (HCF) is a benign fibro-osseous neoplasm of periodontal ligament (PDL) origin containing varying amounts of mineralized material resembling cementum. In the present study, we established cell lines from HCF, which were detected in the mandible of a 54-year-old Japanese man. To obtain immortalized cell clones, we undertook transfection with temperature-sensitive simian virus-40 (SV40) T-antigen and hTERT into HCF cells. Cells transfected with SV40 T-antigen entered "crisis" state between passages 22 and 35, but activation of telomerase by transfection with hTERT in the SV40-transformed HCF cells resulted in bypass of the crisis and maintenance over passage 200. HCF cell lines decreased the expression of SV40 T-antigen and the activity of cell proliferation at a nonpermissive temperature (39 degrees C) in comparison with that at a permissive temperature (33 degrees C). High activities of alkaline phosphatase and mineralization and the expression of type I collagen, osteocalcin, osteopontin, and bone sialoprotein by reverse transcription-polymerase chain reaction (RT-PCR) were observed in HCF cells at 39 degrees C. Overall, these findings suggest that: (i) HCF cell lines may represent a novel in vitro human cell model for the study of the regulatory mechanism of differentiation and proliferation of the human PDL; and (ii) transfection of plasmids encoding the temperature-sensitive SV40 T-antigen gene and hTERT gene may be useful for obtaining immortalized cell lines from benign human tumor and, probably, nonneoplastic human tissues.

Expression of CD44v9 was immunohistochemically studied in 120 biopsy specimens from primary squamous cell carcinoma (SCC) of the tongue and correlated with clinicopathological findings of the SCCs. The tumors were classified into three groups according to immunostaining pattern of CD44v9; 53 cases with distinct positivity in all cancer cells except for those in the central part of nests (Group 1, non-reduced group), 42 cases with reduced expression in peripheral cells of nests (Group 2, reduced group), and 25 cases with complete disappearance of the expression in one or more nests (Group 3, negative group). Nineteen of 25 (76%) tumors in Group 3 and 14 of 42 (33%) in Group 2 exhibited lymph node metastasis, compared with only 8 of 53 (15%) in Group 1. The average survival time in Groups 1, 2 and 3 was 4496±204, 3866±379 and 2719±359 days, respectively and became shorter with the reduction of CD44v9 expression. These results suggest that the down-regulation of CD44v9 in SCC of the tongue may relate to the detachment of tumor cells from primary lesions, establishment of lymph node metastasis and consequently the death of patients.

Down-regulation of p27(Kip1) has been reported to correlate with poor survival of various carcinoma patients including oral squamous cell carcinomas (OSCCs). It is still unclear, however, at what stage of oral carcinogenesis the down-regulation of this protein occurs. In this study, therefore, we evaluated immunoexpression of p27(Kip1) protein in 17 cases of oral epithelial dysplasia and succeeding invasive OSCC in the same patient. We reported here that 88% cases showed high p27(Kip1) expression in dysplastic lesions, whereas 82% cases of succeeding invasive OSCC exhibited reduced expression. The reduction of p27(Kip1) expression was also observed in 16 of 19 (84%) early invasive lesions and well correlated with Ki-67 expression which is good indicator of cell proliferation. We also investigated immunoexpression of p53 protein of which abnormality has been known to occur during the early stage of OSCC development. Overexpression of p53 protein was demonstrated in 29% of dysplastic lesions, 42% of early invasive and 71% of invasive OSCCs. These findings suggest that abnormalities of both p53 and p27(Kip1) are involved in the carcinogenesis of OSCC, but they seem to play their role at different stages of oral cancer development, respectively. Reduced expression of p27(Kip1) may concern the cancer invasion directly or indirectly as well as abnormal proliferation.

To discuss the dental origin of odontogenic myxoma and to provide further information for the differential diagnosis between this tumor and myxoid malignant fibrous histiocytoma (MFH) which occasionally occurs in jaw bones, the contents of glycosaminoglycans (GAGs) and proteoglycans (PGs) in the mucosubstances of 15 odontogenic myxomas, 5 myxoid MFH and 3 human fetal tooth germs in the bell stage of development were characterized using histochemical and immunohistochemical methods. Histochemical staining of hyaluronic acid (HA) was undertaken using biotinylated HA binding protein (B-HABP), and immunohistochemical detection was done using a panel of antibodies against chondroitin 6-sulfate (CS-6), chondroitin 4-sulfate (CS-4), dermatan sulfate (DS), keratan sulfate (KS), heparan sulfate (HS), aggrecan, PG-M/versican, decorin and biglycan. In odontogenic myxoma, CS-6, HA and PG-M/versican were observed in the myxomatous matrix of all cases, while KS and HS were seen in none. As for CS-4, DS, aggrecan, decorin and biglycan, only irregular and mild stainings were shown. Consistent and strong positive straining for CS-6, HA and PG-M/versican were seen in dental papilla and provided evidence supporting the origin of this tumor from dental papilla. Except for the constant staining for HA, the myxoid matrix was rarely stained for most GAGs and PGs in myxoid MFH. Immunodetection of CS-6 and PG-M/version with the use of monoclonal antibodies 3-B-3 and 2-B-1 is therefore recommended as a useful tool in differentiating odontogenic myoma from myxoid MFH.

Detailed three-dimensional morphological information of the cell without distortion due to thin sectioning can be obtained from thick sections using a confocal laser scanning microscope. Here that microscope was used to evaluate human gingival Langerhans cells stained with anti-CD1a antibody. Optical sectioning and reconstruction by laser scanning microscopy revealed not only three-dimensional aspects of Langerhans cells but also spatial information on the distribution of their dendritic processes towards the gingival surface. The orientated configuration of those processes may reflect the reaction of the gingival Langerhans cell to the surrounding moist environment containing antigenic stimuli originating in saliva.

Three cases of monophasic glycogen-rich clear cell tumors of palatal gland origin were examined immunohistochemically and ultrastructurally in attempts to characterize their cellular composition. Despite their histologic resemblances, the clear cells from each case showed different immunohistochemical features. In case 1 the extensive positivity for vimentin and S- 100 protein, in addition to the focal expression of actin and glial fibrillary acidic protein, strongly suggested that the clear cells were myoepithelial in nature. In contrast, the clear cells from case 2 exhibited both keratin and epithelial membrane antigen positivity, as well as ultrastructural features that suggested that they were glandular epithelial in nature. In case 3 no special markers except for keratin could be detected, indicating the less differentiated nature of the clear cells. These results show the heterogeneity of the clear cell tumor group of minor salivary glands.

Four cases of basal cell adenoma of the parotid gland were examined immunohistochemically to characterize their cellular composition. In all cases epithelial membrane antigen and keratin were detected in the inner luminal cells; some cells also showed positive staining for secretory functional markers, indicating their differentiation toward secretory epithelium. In tubular and trabecular types the outer cells consistently displayed an intense staining for vimentin and some were also positive for actin, indicating their myoepithelial nature. In the solid type, most tumor cells resembled the ductal cells or basal cells of larger ducts in normal gland with regard to their immunoreactivity. Our results may suggest that the proportion and arrangement of heterogeneous tumor cells are responsible for different histologic patterns of the salivary basal cell adenoma.

A 4-year-old Japanese girl with a nonpigmented nodule on the anterior portion of the palate since birth is described. The overall microscopic appearance of the lesion was very similar to that of Spitz nevus of the skin. Diagnosis of Spitz nevus (mixed epithelioid cell and spindle cell nevus) was made on the basis of the clinical and histologic criteria for differentiating this lesion from malignant melanomas and common compound nevi. Positive immunostaining of epithelioid and spindle cells for S-100 protein and neuron-specific enolase was also indicative of their nevocellular nature. Review of the cases of Spitz nevus from the literature revealed that the present case most probably represents the first reported instance of this type of nevus in the oral cavity.

Using formalin-fixed and EDTA-decalcified cryostat sections, the immunohistochemical localization of prostaglandin (PG) E2, PGF2 alpha, and 6-keto-PGF1 alpha (a stable metabolite of PGI2) was examined in normal rat and inflamed dental pulp. Inflammation was induced by opening the pulp chamber. There was no immunoreactivity for prostaglandins in normal dental pulp, whereas positivities for PGE2, PGF2 alpha, and 6-keto-PGF1 alpha were demonstrated in the cytoplasm of macrophages and endothelial cells in the inflamed dental pulp. In addition to these cells, numerous pulp cells and odontoblasts existing in the inflamed pulp and its apical noninflamed area also were intensely stained for PGF2 alpha. Such an area with positive cells gradually extended in an apical direction with the progression of inflammation. These findings suggested that PG production from these host cells is involved in development of inflammation of rat dental pulp.

Vascular endothelial growth factor (VEGF) plays a critical role in tumor angiogenesis. VEGF and its receptor FLT-1 signaling may contribute to cell migration of macrophages, osteoclasts and some tumor cells. However, the direct role of VEGF-Flt-1 signaling in malignant behaviors of oral squamous cell carcinoma (OSCC) is not well understood. To clarify the effects of VEGF-Flt-1 signaling in OSCC bone invasion, VEGF expression in OSCC cells and number of Flt-1+ osteoclasts at bone invasion front in 55 gingival OSCC cases were evaluated by immunohistochemistry. Twenty-four of 55 cases (43.63%) strongly expressed VEGF. The VEGF highly expressing cases were statistically correlated with the increased number of Flt-1+ osteoclasts and the more aggressive radiographic pattern of bone invasion (p In conclusion, VEGF-Flt-1 signaling promotes osteoclastogenesis, proliferation and facilitating invasion and migration of OSCC through production of MMP-1 and 3. Blocking of VEGF-Flt-1 signaling may be beneficial for OSCC treatment.