Yutaka Kimijima

Aetiology of maxillofacial fracture

The aetiological factors associated with maxillofacial fractures, and the trends in these factors over a 13 year period are reported. The First Department of Oral and Maxillofacial Surgery, Faculty of Dentistry, Tokyo Medical and Dental University, managed 695 patients with maxillofacial fractures between 1977 and 1989. The male to female ratio was 3.2:1 and the majority of patients were aged between 10 and 30 years old. Road traffic accidents and accidental falls were the main causes of fractures throughout the 13 year study period. Mandibular fractures occurred in 477 patients (68.6%). A high percentage of patients were treated by closed reduction and maxillo-mandibular fixation, or occlusal splinting.

Maxillofacial fractures in children

A clinico-statistical and long-term follow-up study was performed on 81 pediatric fractures seen during the 14 years between 1977 and 1990. Of all maxillofacial fractures, the incidence of pediatric fractures was 14.7%. The ratio of boys to girls was 2.1:1, and the highest incidence involved boys over 13 years of age. Fractures of the upper alveolar bone and mandible were common. Conservative therapy, such as maxillomandibular fixation using orthodontic brackets was usually performed and was found to be successful. The long-term follow-up study revealed that 5 out of 21 patients with alveolar fractures complained of malocclusion and it is suggested that a longer duration of intramaxillary fixation and long-term follow-up might be needed for alveolar fractures in children.

Expression of Rb, pRb2/p130, p53, and p16 proteins in malignant melanoma of oral mucosa

We previously reported that pRb2/p130 gene, one of the Rb family members, was immunohistochemically abundantly expressed in well-differentiated oral squamous cell carcinomas, whereas in undifferentiated ones the expression was low. Oral malignant melanoma is extremely rare, however the prognosis is poor because it tends to locally invade tissue or metastasize and its biological behavior appears to be different from cutaneous malignant melanoma. The present study dealt with the expression of pRb2/p130, Rb, p53, and p16 in 13 cases of malignant melanoma of oral mucosa as revealed by immunohistochemical staining. The stage classification of the 13 patients was as follows; stage II: eight patients, stage III: three patients, and stage IV: two patients. pRb2/p130 was expressed in only two stage II-cases, neither of which have shown any evidence of recurrence or metastasis for over 14 years. Positive staining for Rb was found in three cases consisting of one stage II-case, one stage III-case, and one stage IV-case. p53 was expressed in two cases, one a stage II and the other a stage IV. Positive staining for p16 was found in seven cases consisting of four stage II-cases, two stage III-cases, and one stage IV-case. pRb2/p130 may be inversely correlated with the malignancy of oral malignant melanoma, but further study is needed.

Possible etiology of calculi formation in salivary glands : biophysical analysis of calculus

Sialolithiasis is one of the common diseases of the salivary glands. It was speculated that, in the process of calculi formation, degenerative substances are emitted by saliva and calcification then occurs around these substances, and finally calculi are formed. However, the exact mechanism of the formation of calculi is still unclear. In this study, we identify some possible etiologies of calculi formation in salivary glands through biophysical analysis. Calculi from 13 patients with submandibular sialolithiasis were investigated by transmission electron microscopy, scanning electron microscopy, X-ray microanalyzer, and electron diffraction. Transmission electron microscopic observation of calculi was performed in the submandibular gland (n = 13). In 3 of the 13 cases, a number of mitochondria-like structures and lysosomes were found near calcified materials. Scanning electron microscopic examination of these materials revealed that there were lamellar and concentric structures and that the degree of calcification was different among the calculi. X-ray microanalysis disclosed the component elements in the calculi to be Ca, P, S, Na, etc., and the main constituents were Ca and P. The calcium-to-phosphorus ratio was 1.60–1.89. Analysis of the area including mitochondria-like structures, lysosomes, and the fibrous structures by electron diffraction revealed the presence of hydroxyapatite and calcified materials. It is speculated that mitochondria and lysosomal bodies from the ductal system of the submandibular gland are an etiological source for calcification in the salivary gland.

Immunohistochemical investigation of new suppressor oncogene p130 in oral squamous cell carcinoma

Abstract New onco-suppressor gene p130 is one of the Rb family forms and is reported to undergo allelic loss in hepatocellular, prostate, and breast carcinomas; however, no report of p130 has been made in oral tumors. The present study dealt with the expression of p130 and Rb proteins by immunohistochemical staining in oral squamous cell carcinomas ( n =110) and oral mucosa. The site of the carcinomas included tongue ( n =48), gingiva ( n =32), oral floor of the mouth ( n =15), oropharynx ( n =6), buccal mucosa ( n =5), and others ( n =4). Histologically there were 65 well-differentiated carcinomas, 33 moderately differentiated carcinomas, and 12 poorly differentiated ones. Positive staining for p130 and Rb was localized to suprabasal cell layers of the normal oral epithelium. In the oral squamous cell carcinoma, positive staining for p130 and Rb was observed in well-differentiated carcinomas (p130, 66.2%; Rb, 78.5%), more than in poorly differentiated ones (p130, 16.7%; Rb, 50.0%). In immunoelectron microscopic features, p130 protein was localized in the nucleus and mitochondria. The expression of p130 was related to the degree of tumor differentiation as that of Rb. It is suggested that p130 gene may be associated with the development of a wide variety of human malignancies rather than the progression.

Ultrastructural analysis of salivary calculus in combination with X-ray microanalysis

 Ultrastructural studies of salivary calculi were performed. Scanning electron microscopic examination of the calculi revealed lamellar and concentric structures. Granular or globular structures and pyramid structures were found on the surface of the calculi, and in some cases a scaly structure corresponding to fiber and bacteria was recognized. X-ray microanalysis showed the main constitutes of the calculi to be Ca and P. Transmission electron microscopic examination revealed a fine fibrous structure near the degenerated organelles, and analyses of the structure by electron diffraction revealed hydroxyapatite. Calcification was found around the degenerative organelles in the form of lipid-like structures, mitochondria, lysosomes, and microbial structures. Judging from our results, as one of the processes leading to calculi formation, it is speculated that degenerative substances are emitted by saliva, by some phenomenon, and calcification around these substances then occurs, contributing to the formation of calculi.

Oral squamous cell carcinoma: electron microscopic and immunohistochemical characteristics

 Oral squamous cell carcinoma is the most common oral malignancy, and we performed electron microscopic and immunohistochemical investigation of the tumor. In patients with cervical metastasis, microvilli were developed and a small number of desmosomes were found, regardless of the width of the intercellular spaces. In patients without the metastasis, few microvilli were found in relatively wide intercellular spaces, or numerous microvilli were found in narrow intercellular spaces, and a large number of desmosomes were shown. However, these findings were different from those of tumors that had received radiotherapy, in which numerous microvilli and a small number of desmosomes were found in the nonmetastatic cases. Transferrin receptor, which is a marker of cell proliferation, was localized on the cell membrane, especially in microvilli. Ultrastructural similarity between the primary tumor and the metastatic tumor was recognized, however, the features of microvilli, desmosomes, and the intercellular spaces differed between them in most cases. It is suggested that microvilli might be related to the metastasis of oral squamous cell carcinoma cells. Immunohistochemically, the protein expression of p53 and pRb2/p130 was related to the clinical course of the patients with oral squamous cell carcinoma; the mechanism of the synthesis of these proteins should be investigated in order to understand the biological behavior of the tumor.

Investigation of titanium leak to bone tissue surrounding dental titanium implant : electron microscopic findings and analysis by electron diffraction

This study investigated the tissue response associated with dental titanium implants. The mandibular third and fourth premolars and first molar of three adult beagle dogs were extracted bilaterally. Healing was then allowed for 3 months. Six titanium implants were placed in the mandibles of a dog. Three weeks after the implantation, mandibular sections containing the implants were retrieved with the use of a bone saw and investigated by light and electron microscopy, X-ray microanalyzer, and electron diffraction. Scanning electron microscopic observation showed titanium particles on the implant–bone interface, and investigation by microanalyzer revealed titanium not only on the implant–bone interface but also in the bone tissue. Transmission electron microscopic observation and investigation by electron diffraction showed titanium in the bone matrix and cells other than macrophages. In this study, titanium particles from the dental implant were recognized morphologically in the surrounding bone tissue. Thus, study of the influence of titanium particles on the human body is needed.

An ultrastructural study of the influence of radiation against cell line-derived human oral malignant melanoma

The response to radiation of the SSM-1 cell line, derived from oral malignant melanoma, was investigated by survival curve and by light and electron microscopic abservation. Survival curve of the cells was investigated by the colony formation assay. X-irradiation with 2, 4, and 8 Gy was performed against the cell line. Morphological changes of the cells at 6, 24, 72, and 120h after irradiation were examined by both light and electron microscopy. The survival curve of the SSM-1 cells showed higher radiosensitivity than that of cutaneous melanoma cell lines. Giant cells and multinucleated cells were found only 120h after 8Gy irradiation. Ultrastructural observation revealed changes in nucleus and melanosomes; melanosomes increased in number at 120h after 8Gy irradiation. Further alterations after irradiation were noticed primarily in the nucleus. Radiosensitivity of the SSM-1 cell line derived from oral malignant melanoma was higher than that of cutaneous melanoma cell lines. The results of this study may support the concept that radiotherapy is effective for oral malignant melanoma.

An ultrastructural study of calcifying odontogenic cyst, especially calcified material

 The ultrastructural features of calcification in a case of calcifying odontogenic cyst (COC) were studied. Scanning electron microscopic examination of the inner parts of the cyst wall revealed many short microvilli, and X-ray microanalysis of the high-density masses in the intercellular parts showed prominent calcium peaks, which meant that these masses were calcified materials. On transmission electron microscopic observations, many calcifications exhibited a distinctive ring formation around the periphery of a central core that consisted of an amorphous structure. These calcifications were observed with necrotic remnants of nuclear material and many identifiable mitochondria, thin fibers, and epithelial cells. The cytoplasm of ghost cells consisted of numerous short electron-dense tonofilament bundles. Needle-like structures were shown in the tonofilament bundles. X-ray diffraction analysis showed that the needle-like crystals were hydroxyapatite. It is suggested that calcification in a COC may be related to degenerative mitochondria and tonofilament bundles of ghost cells.