Characterization of mesenchymal progenitor cell populations from non-epithelial oral mucosa.

OBJECTIVES: The characteristics of cell populations extracted from oral mucosal non-epithelial tissues and their ability to differentiate were evaluated in vitro as a potential source of cells for mandibular and corneal regeneration. MATERIALS AND METHODS: Oral mucosal non-epithelial cells (OMNECs) were extracted from tissue samples and were studied by flow cytometry and RT-PCR. Cells differentiating into osteoblasts, adipocytes, chondrocytes, neurocytes, or keratocytes were characterized by RT-PCR and cell staining. RESULTS: OMNECs expressed CD44, CD90, CD105, CD166, and STRO-1 antigens, which are markers for mesenchymal stem cells. In addition, Oct3/4, c-Myc, Nanog, KLF4, and Rex, which are expressed by embryonic or pluripotent stem cells, were detected by RT-PCR. Expression of CD49d, CD56, and PDGFRα, proteins closely associated with the neural crest, was observed in OMNECs, as was expression of Twist1, Sox9, Snail1 and Snail2, which are early neural crest and neural markers. Specific differentiation markers were expressed in OMNECs after differentiation into osteoblasts, adipocytes, chondrocytes, or keratocytes. CONCLUSIONS: Populations of OMNECs may contain both mesenchymal stem cells and neural crest origin cells and are a potential cell source for autologous regeneration of mandibular or corneal stroma.

Three-dimensional finite elemental analysis of zygomatic implants in craniofacial structures.

The objective of this study was to analyse stress distribution in craniofacial structures around zygomatic osseointegrated implants. An integrated system for Digital Imaging and Communications in Medicine (DICOM) data were utilized to create a three-dimensional model of craniofacial structures. The amount and distribution of the main stresses were compared using three-dimensional finite elemental analysis. The system allowed visual confirmation and analysis of stress distribution as well as the convenient and simple construction of a digital biomechanical model that provided details of anatomical structures in the regions of interest. Zygomatic implants with or without connected implants supporting the superstructure were compared. Stresses in severely resorbed maxillae with connected implants were not concentrated around the alveolar bone supporting the zygomatic implants. Stresses where there were no connected implants tended to be generated in the zygomatic bone, at the middle part of the zygomatic implant and at the joint of the fixture-abutment. Stress due to occlusal forces is mainly supported by the zygomatic bone, is transferred predominantly through the infrazygomatic crest, and is divided between the frontal and temporal processes of the zygomatic bone in different directions.

Investigation of environmental factors for diagnosing malignant potential in oral epithelial dysplasia.

A study was conducted in rats with early tongue carcinoma induced by 4-nitroquinoline 1-oxide (4NQO), in order to investigate the early diagnosis of malignant potential of epithelial dysplasia. The rat tongue lesions were classified by their severity into three groups corresponding to early cancer, dysplasia and no change. The grade of epithelial changes was determined according to 13 items of WHO Epithelial Dysplasia Criteria. The expression levels of p53 and Bcl-2 proteins were detected immunohistochemically, and apoptotic cells were detected using the TUNEL method. In addition, a p53 mutation by lesions was detected. The expression ratio of p53 protein was high in dysplasia, and the ratio of Bcl-2 protein was high in early cancer and dysplasia. The TUNEL-positive cells were observed primarily in the granular layers of the no change cells, and their numbers decreased as the cells shifted to the early cancer stage. The p53 mutation was detected using a microdissection method in dysplasia, where it was found in three out of nine lesions. All the mutations in dysplasia detected were on the same codon that was found to be mutated in the early cancer. These results indicate that the association between the p53 mutation and histological changes in carcinogenesis epithelial dysplasia is strong, and that both the identification of p53- and Bcl-2-positive epithelium, and decrease in the TUNEL positive ratio, were useful for the diagnosis of the malignant potential of precancerous lesions.

The carcinostatic effects of 1-(2-tetrahydrofuryl)-5-fluorouracil and uracil (UFT) on tongue carcinoma induced by 4-nitroquinoline 1-oxide (4NQO) in rats.

UFT is a carcinostatic agent used in adjuvant chemotherapy for head and neck cancer. In the present study. UFT was given orally to treat tongue carcinoma in rats induced by 4-nitroquinoline 1-oxide. The antitumor effects of UFT were studied macroscopically and histologically. In addition, the antitumor effects of UFT were evaluated in relationship to lesions of the clinical and, invasive types, and to there vascular structure. In clinical lesions, the antitumor effect of UFT was higher in extrovert tumor-mass lesions and lower in ulcerous lesions. With regard to vascular structure, the effect was higher in cases demonstrating irregular net-like patterns and branch-like patterns and lower in cases in which the pattern had been destroyed. There was a correlation between antitumor effect and invasive type. As invasive tendency the 3H-thymidine labeling index, and mitotic index increased, antitumor effect and degree of tumor cell degeneration decreased.

Microvascular free tissue transfer in oral and maxillofacial reconstruction.

Forty-two patients with head and neck cancer were submitted to microvascular reconstructive procedures. We divided patients in three groups; a first group of 30 patients, in whom the oral floor (8 patients), the part of tongue (14 patients), the lower gingiva (6 patients) and the oropharynx (2 patients) were reconstructed using various sizes of forearm flaps; a second group of 7 patients who underwent buccal mucosa reconstructions with the forearm flaps; and a third group of 5 patients who received rectus abdominis flaps for total tongue reconstruction. Three illustrative cases, one from each group, are presented in detail. Good results were obtained in 39 patients (94%), with both functional and morphological rehabilitation. There were three flap losses due to thrombosis of the microvascular anastomoses. There was no surgical mortality. The average operating time was about 10 hours in total. We concluded that there is a place for these complex procedures in the treatment of selected cases of head and neck tumors.

Clinicostatistical study of ameloblastoma treatment.

The purpose of this study was to investigate the treatment of 190 cases of ameloblastoma in our department from 1966 to 1994. The statistical results with regard to age, sex and region agreed with those of other investigators. Thirty-five of 43 (81.4%) cases underwent enucleation in 1960s, but the sixteen of 27 (59.3%) cases underwent partial resection of mandible in 1990s. The defect of mandible was reconstructed with iliac bone grafting since 1968, grafts with a mixture of iliac blocked bone and PCBM (particulate cancellous bone and marrow) have been used since 1975. Grafting of the inferior alveolar nerve with the great auricular nerve to the defect has been performed in our department since 1977. Recently, technique involving pull-through of the inferior alveolar nerve bundle has been used in our department. When the reconstruction method for the mandible and nerve has been established, it becomes possible to operate radically and positively. Recurrence occurred in 17 cases after the primary enucleation. It is thought that the primary treatment of ameloblastoma must be as radical as possible. It appears to be necessary to observe progress and perform follow-up in cases of ameloblastoma for more than ten years, because there was one recurrence at 9 years and 4 months after the first operation. In fact, three quarters of our cases were lost to follow-up. Such losses can problems in confirming recurrence and responding rapidly.

Biomechanical three-dimensional finite-element analysis of maxillary prostheses with implants. Design of number and position of implants for maxillary prostheses after hemimaxillectomy.

The present study analyzed stress distributions in craniofacial structures around implant-supported maxillary prostheses. Using post-hemimaxillectomy computed tomography (CT) of a patient, the authors constructed a three-dimensional (3D) solid model using Digital Imaging and Communications in Medicine data (DICOM data) for maxillofacial and cranial bones. The effects of different prosthesis designs on stress distributions in craniofacial bones and osseous tissues around the implants were biomechanically investigated using 3D finite-element analysis. Maxillary prostheses were designed with 2 implants in the zygoma on the affected side and 2-3 implants in the maxillary alveolar bone on the unaffected side, without using a cantilever. Zygomatic implants provided suitable stress dispersal to the zygomatic and craniofacial bones on the affected side. This information is useful for designing maxillary prostheses.

Isolation of oral epithelial progenitors using collagen IV.

OBJECTIVE: Although oral mucosal epithelial stem cells are thought to reside in the basal layer, such cells have not yet been isolated. We isolated a population of rabbit oral epithelial progenitor cells containing putative stem cells. MATERIALS AND METHODS: Epithelial cells harvested from rabbit buccal mucosa were allowed to adhere to dishes coated with collagen IV for periods ranging from 10 min to 16 h. The properties of individual cell populations were evaluated using BrdU, Ki-67, integrin beta1, integrin alpha6 and keratin 13 using colony forming efficiency (CFE). RESULTS: Cells that adhered to collagen IV-coated dishes within 10 min were enriched about sixfold in terms of BrdU incorporation, Ki-67, integrin alpha6 and integrin beta1 were strongly expressed. Interestingly, keratin 13 was faintly expressed. The CFE of rapidly adherent cells among oral epithelial cells was significant compared with other cell populations. CONCLUSIONS: These results suggested that rabbit oral epithelial cells could be isolated by depending on adhesiveness to collagen IV, especially when segregated according to progenitor cell properties. Putative progenitor cells with stem cell properties were most effectively harvested within 10 min. Our separation procedure should be a useful tool with which to isolate epithelial stem cells for regenerative medicine.