Applicability of human dental follicle cells to bone regeneration without dexamethasone: an in vivo pilot study.

The aim of this study was to investigate the capacity of human dental follicle cells (hDFCs) for bone formation in vivo. hDFCs were obtained from wisdom teeth extracted from patients aged 14 and 22 years. hDFCs from the 5th to 8th passages were grown in three-dimensional (3D) culture using gelatin sponges. Cells were transplanted onto the calvaria of F344/NJcl-rnu/rnu male rats (immunodeficient rats). Haematoxylin and eosin (HE) staining and immunohistochemistry were performed, and newly formed bone was evaluated by micro-computed tomography (micro-CT). HE staining showed newly formed bone in 3D culture. Immunohistochemistry showed bone morphogenetic protein 2 (BMP-2), runt-related transcription factor 2 (RUNX2), and osterix staining in areas with newly formed bone. Furthermore, micro-CT showed that, in comparison to controls, transplanted hDFCs promoted better bone quality and bone mineral density (BMD 582 ± 131.1 vs. 300.5 ± 77.7 mg/cm(3); P=0.039), bone mineral content (BMC 5.6 ± 1.1 vs. 2.1 ± 0.4 mg; P = 0.006), bone volume (BV 9.7 ± 0.5 × 10(-3) vs. 7.0 ± 0.4 × 10(-3) cm(3); P = 0.002), BMC/total volume (TV) (399.9 ± 76.3 vs. 147.7 ± 30.8 mg/cm(3); P = 0.006), and BV/TV (69.1 ± 3.6% vs. 49.6 ± 3.1%; P=0.002). This suggests that human dental follicles are potentially useful for regenerative therapy.

MCP-1 production in temporomandibular joint inflammation.

Synovitis, which is characterized by the infiltration of inflammatory cells, often accompanies progression of temporomandibular joint disorder (TMD) symptoms. Because IL-1ß is elevated in synovial fluids obtained from TMDs, we hypothesized that IL-1ß-responsive genes in synoviocytes may help identify the putative genes associated with synovitis. Using microarray analysis, we found that monocyte chemoattractant protein-1 (MCP-1) mRNA levels were elevated in IL-1ß-stimulated synoviocytes. MCP-1 is a member of the chemokine superfamily. The production of MCP-1 was increased in synoviocytes treated with IL-1ß. When IL-1ß was injected into the cavities of rat TMJs, inflammatory cells and MCP-1-positive cells were detected in the synovial tissues. Furthermore, MCP-1 levels were higher in synovial fluids from individuals with pain compared with those without pain. Inhibitors of MAP-kinases and NF-κB reduced IL-1ß-induced MCP-1 production. These results suggest that MCP-1 stimulated by IL-1ß is one of the factors associated with the inflammatory progression of TMDs.

Expression of cyclooxygenase-1 and -2 in IL-1beta-induced synovitis of the temporomandibular joint.

BACKGROUND: In this study, we analyzed the gene expression profile of fibroblast-like synoviocyte (FLS) cultures from the temporomandibular joint (TMJ) to identify candidate genes associated with intracapsular pathologic conditions of TMJ. Cyclooxygenase (COX)-2 was one of the genes in FLS upregulated following stimulation by interleukin (IL)-1beta, a cytokine thought to play a key role in several pathological conditions. This study investigated the expression of COX-1 and COX-2 in cultured human FLS and rat TMJ synovium following stimulation with IL-1beta. METHODS: RNA was isolated from human FLS after IL-1beta treatment. COX-1 and -2 expression was examined using a GeneChip and real-time polymerase chain reaction. Prostaglandin E(2) (PGE(2)) levels in conditioned media from FLS were measured using enzyme-linked immunosorbent assay. Synovial tissues from TMJs of IL-1beta-injected rats were examined for COX-1 and COX-2 expression by immunohistochemical staining. RESULTS: Following treatment of FLS with IL-1beta, expression of the COX-2 gene increased up to 8 h and peaked at 4 h, whereas COX-1 expression did not change. Stimulation with IL-1beta increased the level of PGE(2) in conditioned media of cultured FLS in a time-dependent manner up to 48 h. Immunohistochemistry showed a strong positive staining for COX-2 in the lining and sub-lining synovial tissues of the TMJ of IL-1beta-injected rats. In contrast, staining for COX-1 was the same in synovial tissues with and without IL-1beta injection. CONCLUSION: These data suggest that COX-2 expression stimulated by IL-1beta stimulates the production of PGE(2) in FLS and plays important roles in the progression of inflammation in TMJ.

Distribution of basal ganglia lesions in generalized variant of Pick's disease: a clinicopathological study of four autopsy cases.

We investigated four Japanese autopsy cases of the generalized variant of Pick's disease ("basophilic inclusion body disease") both clinically and pathologically, and examined the degree and distribution of the basal ganglia lesions, including the amygdala, striatum, pallidum, and substantia nigra. The lesions in the amygdala, striatum, and pallidum were classified into three categories (slight, moderate, and severe). The lesions in the substantia nigra were qualitatively judged, compared with normal controls. Extrapyramidal signs, not noticed in the generalized variant of Pick's disease, were evident in all four cases, in addition to dementia. The degree and distribution of basal ganglia lesions in all four cases were uniform: the caudate nucleus showed severe lesions, the amygdala and putamen severe to moderate lesions, and the pallidum moderate to slight lesions. The substantia nigra in all our cases showed prominent neuronal loss, probably being one of the lesions responsible for extrapyramidal signs. In the generalized variant of Pick's disease, the degree and distribution of the alterations within the basal ganglia differs from those reported in Pick's disease with Pick bodies (PDPB) and corticobasal degeneration (CBD). In PDPB, severe lesions are present in the amygdala with relative sparing of the substantia nigra, compatible with rare extrapyramidal signs in PDPB, while in CBD, severe lesions are found in the pallidum and substantia nigra. These clinicopathological findings may contribute not only to the elucidation of clinicopathological hallmarks, but also to the progress of neuroimaging, in the generalized variant of Pick's disease.

Comparison of occurrence of oro-maxillo-facial tumor types in different regions of the People's Republic of China.

The frequencies of oro-maxillo-facial tumors, by type, seen at the Guangxi Medical College Hospital were surveyed in order to compare different regions of the People's Republic of China. Computer analysis was performed on data for all oro-maxillo-facial tumors, as confirmed by the hospital between 1957 and 1987, inclusive. The results were then used in a comparative study with those of five other medical colleges. Among a total of 4,052 cases, there were 1,593 benign tumors, 2,049 malignant tumors and 410 unknown tumors. Tumors derived from the epithelium were the most common type (42.0% of the total), whereas in the other five medical colleges the mean corresponding proportion was 27.8%, the difference being significant (p < 0.05). Epithelial tumors were also the most common type of malignant tumor (70.5% of the total), whereas the corresponding proportion reported by the other five institutions was 60.6%, which was also significantly different (p < 0.05). The proportion of malignant tumors among the total was more than 50.6%, which was higher than that reported by the other five colleges. This study shows that epithelium-derived tumors and malignant tumors were encountered more frequently at Guangxi Medical College than at the other five medical colleges in China.

Myoepithelioma arising from the buccal gland: histopathological and immunohistochemical studies.

A rare case of myoepithelioma of the buccal gland in a 54-year-old Japanese woman is reported. As the swelling exhibited a normal mucosal color and was relatively well defined, showing no ulcers, a benign salivary gland tumor was suspected upon clinical inspection. Microscopically, the parenchyma of the present case mainly consisted of plasmacytoid cells with round nuclei and eosinophilic cytoplasm, and partial spindle cells with eccentric nuclei. The stroma was composed of fibro-hyalinized or myxoid connective tissue that separated from the parenchyma. Immunohistochemically, the cytoplasm of the plasmacytoid and spindle cells was moderately positive for vimentin and GFAP, whereas the buccal gland adjacent to the tumor was negative for these antibodies. S-100 protein reactivity is strong for both types tumor cells. Actin reactivity was negative for both types of tumor cells, notwithstanding the fact that myoepithelial cells of the buccal gland were positively stained. Anti-cytokeratin reactivity was weak for both types of tumor cells in portions of the plexiform and solid areas; nevertheless, the buccal glands were moderately positive. These results suggest that neoplasmic myoepithelial cells exhibit abnormal differentiation and modification. There have been only two published reports of myoepithelioma arising from the buccal gland in the literature to date.

Acinic cell carcinoma of the palate: case report and immunohistochemical observation.

We report a rare case of acinic cell carcinoma of the palate in a 63-year-old Japanese woman. Clinical examination demonstrated a firm, mobile mass without regional lymph-adenopathy. Histopathologically, the tumor was composed of large, polyhedral or round cells with basophilic granular cytoplasm (serous acinar-like cells) and reticular or clear cytoplasmic cells. These tumor cells were positive for the periodic acid-Schiff reaction, but negative for alcian blue. The tumor nests were separated by thin vascular tissue and incompletely encapsulated. Immunohistochemically, the tumor cells exhibited positive reactivity for alpha-amylase, lactoferrin, secretory component, S100 protein, and epithelial membrane antigen, but were negative for actin, glial fibrillary acidic protein, keratin, and carcinoembryonic antigen. These results suggest that this tumor is well differentiated into serous acinar cells and that the reticular and clear cytoplasmic cells are a modified form of these cells.

Crystallization and preliminary X-ray analysis of a truncated family A alkaline endoglucanase isolated from Bacillus sp. KSM-635.

The catalytic domain of an alkaline endo-1,4-beta-glucanase (family A) isolated from Bacillus sp. KSM-635 (Mr = 40.2 kDa) was crystallized using the hanging drop vapor diffusion method. Two different crystal forms were obtained. Form 2 crystals (trigonal space group R3 with cell dimensions of a = b = 111.9 and c = 207.1 angstroms in a hexagonal lattice) were found to be more stable than form 1 ones upon X-ray irradiation. A full data set for form 2 crystals has been collected up to 3.3 angstroms resolution.

A study of effects of mouthwash on the human oral mucosae: with special references to sites, sex differences and smoking.

In recent years, the use of mouthwash has become widespread as a part of routine oral hygiene. However, there have been no fundamental studies on the influence of mouthwashes on the human oral mucosae. One hundred and twenty-five subjects (50 males and 75 females) were selected for this study. The effects of mouthwash was assessed with the use of exfoliative cytological and cytomorphometric analyses of smears obtained from clinically normal upper labium and cheek mucosae before mouthwashing, 30 s, 10 min and 1 h after mouthwashing. The independent variables examined were oral site, sex and smoking (smokers versus never-smokers). In all subjects and sites, the appearance rate of exfoliated cells stained by light green SFY decreased just after mouthwashing, and the rate after 1 h was lower than that of the untreated controls. The oral mucosae of smokers were more irritated action by mouthwashing than that of the never-smokers. Even after 1 h, decreases in the nuclear and cytoplasmic areas of cells and increases in inflammatory cells were observed. In conclusion, the use of mouthwash was so inflamed the human oral mucosae that more attention should be paid when it is used daily in oral hygiene.

An autopsy case of spinal arteriovenous malformation (Foix-Alajouanine syndrome).

An autopsy case of spinal arteriovenous malformation (AVM) was reported. The patient was a 75-year-old male and his initial neurologic symptoms were paraplegia, paresthesia below the umbilical level and urination difficulty. Subsequently night delirium and parkinsonism also appeared. The clinical and pathological findings in this case are identical with those in the spinal AVM except for Parkinson's disease. In addition, the lateral funiculus of the spinal cord in the middle thoracic segment showed pallor: Under light microscopy, the funiculus was spongiform, with a thinner wall of the myelin sheath, enlargement of the axon and the perivascular infiltration of phagocytes without plasma exudation. The changes in the lateral funiculus seemed to indicate early congestive changes.

[An autopsy case of multiple system atrophy with many Lewy bodies--striatonigral degeneration, olivo-ponto-cerebellar atrophy and autonomic nerve nucleus involvement in the spinal cord].

A 72 years old man developed slowness of the motion and orthostatic hypotension at the age of 69. Neurological examination showed slight finger tremor, rigidity of extremities, bradykinesia, and marked orthostatic hypotension. The illness progressed steadily and the patient died of pneumonia. At autopsy brain weighed 1220 g. Grossly the putamen was bilaterally shrunken, the color of the substantia nigra and locus ceruleus became pale. Base of the pons and the cerebellum were atrophic. Microscopically the most remarkable change was seen in the striato-nigral system. In the putamen, there were severe loss of small neurons and intense gliosis and brownish pigments were observed in the neuropil and within some of the astrocytes. There found neuronal loss and gliosis in the substantia nigra. A few Lewy bodies were seen in the substantia nigra. In the cerebellum there were slight loss of Purkinje cell and many torpedos were seen. There were demyelination and fibrirally gliosis in the cerebellar white matter except the hilus of dentate nucleus. The transverse pontocerebellar fibers were degenerated and fibrirally gliosis was seen there. The inferior olivary nuclei showed neuronal loss and astrocytosis. But the degeneration of the olivo-ponto-cerebellar system in this case was not so severe as the typical case of OPCA. In the spinal cord there was depletion of nerve cells in the intermediolateral nuclei and Onufrowitz nuclei. Slight neuronal loss and many spheroids were observed in the anterior horns and there was demyelination in the corticospinal tracts.(ABSTRACT TRUNCATED AT 250 WORDS)

[An autopsy case of amyotrophic lateral sclerosis with dementia and neck extension].

A 58-year-old man recognized hoarseness and dysarthria followed by weakness of the left upper extremity. Examination five months later disclosed dementia which was manifested dominantly by changes in personality and behavior, but also by symptoms of amyotrophic lateral sclerosis. He became progressively weak, and his neck became rigid and extended for two months. He expired due to respiratory failure at the age of 60. At autopsy, the brain weighed 1120 g with atrophy of the temporal lobes. Microscopic examination revealed marked diminution of Betz cells with astrocytic proliferation in the motor cortex. There was a mild, localized spongy state in the upper layer in the frontal and temporal cortices. There were very few neurofibrillary changes and senile plaques, and no Pick's argentophilic bodies. Fibrillary gliosis was found in the white matter of the temporal lobes, external segment of the globus pallidus and the amygdaloid nucleus. The substantia nigra showed depletion of pigmented cells, free melanin pigment and reactive astrocytosis. A diminished number of motor neurons in the brain stem and spinal cord accompanied astrocytic gliosis, while the remaining cell contained many Bunina bodies. The pyramidal tracts showed mild degeneration bilaterally below the pyramis in the medulla. There are a number of cases in the literature whose pictures consist of amyotrophic lateral sclerosis, dementia and/or extrapyramidal symptoms. This case is identical to those cases. But in this case, the clinical and pathological features of amyotrophic lateral sclerosis are more dominant than in other cases.