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.

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.

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.

Langerhans' cells in the murine oral mucosa in the inductive phase of delayed type hypersensitivity with 1-chloro-2, 4-dinitrobenzene.

We created a murine model of delayed-type hypersensitivity (DTH) to 1-chloro-2, 4-dinitrobenzene (DNCB). Using this murine model, we compared oral mucosal sensitization and skin sensitization for the difference in reaction during the elicitation phase. Evaluation of sensitizability, using the mouse ear swelling test (MEST) after oral mucosal or skin sensitization, showed that the ear swelling response peaked 24 h after challenge. The optimal induction concentration was 1.0% in both oral mucosal and skin sensitization, resulting in a positive reaction rate of 100%. However, the ear swelling response 24 h after challenge with the optimal concentration of DNCB (1.0%) was significantly lower in oral mucosal than in skin sensitization. We compared the oral mucosal and skin sensitization sites for the number of Langerhans' cells (LC) and the antigen-presenting capability in the induction phase. The numbers of F4/80+ major histocompatibility complex (MHC) class II+ LC before induction did not differ significantly between the oral mucosa and the skin. After induction, F4/80+ MHC class II+ LC increased in number, but the increase was significantly smaller in the oral mucosa than in the skin. MEST on anti-CD86 antibody-administered mice showed that ear swelling was similarly suppressed after oral mucosal or skin sensitization. In murine models of DTH after oral mucosal sensitization, the number of F4/80+CD86+ LC increased after induction, but the increase was significantly smaller than that in murine models of DTH after skin sensitization. This study showed that, in murine models of DTH, oral mucosal sensitization elicited a weaker reaction than skin sensitization. This was presumably because oral mucosal sensitization induced fewer LC, resulting in lower antigen-presenting capability.

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.

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.

Relationships between the onset of pustulosis palmaris et plantaris, periodontitis and bacterial heat shock proteins.

Relationships between the onset of pustulosis palmaris et plantaris, periodontitis and heat shock proteins were studied by using enzyme-linked immunosorbent assay to examine levels of immunoglobulin G (IgG) against Escherichia coli GroEL, a recombinant DnaJ of Actinobacillus actinomycetemcomitans heat shock protein, a synthetic peptide made from the 180th to the 188th amino acids of Mycobacterium bovis BCG Hsp65, and a recombinant human Hsp60, in sera obtained from 43 pustulosis palmaris et plantaris patients judged to have chronic infectious diseases of the oral cavity. We found that the titers of IgG against E. coli GroEL and A. actinomycetemcomitans DnaJ in the sera from pustulosis palmaris et plantaris patients were significantly higher than those in the control group, whereas the titers of IgG against the synthetic M. bovis Hsp65 and the recombinant Hsp60 did not differ significantly. Periodontal therapy and extraction of teeth with periapical infectious resulted in remission of pustulosis palmaris et plantaris and a statistically significant reduction in the levels of IgG against E. coli GroEL in 9 of the 22 patients (41%) examined. We also found that the IgG levels against A. actinomycetemcomitans DnaJ in 6 serum samples of 16 (37%) were reduced, but not significantly, after the treatment. These results suggest that the IgG responses to heat shock proteins partially induced by oral bacteria may be related to the onset of pustulosis palmaris et plantaris in some patients.

Accidental injection of a U.S. Air Force aviator by a pralidoxime chloride auto-injector: a case report.

To counter the threat of organophosphate nerve agents, military personnel may be issued auto-injectors containing pralidoxime chloride. This drug helps to dephosphorylate the nerve agent-acetylcholinesterase complex and, thus, regenerate the enzyme. In non-poisoned persons, pralidoxime chloride is rapidly excreted by the kidneys and is fairly well tolerated. We present the first reported case of an accidental injection of an Air Force aviator by an auto-injector. The patient recovered well with no specific treatment needed. The pharmacology and toxicology of pralidoxime chloride are discussed.

A clinical analysis of the recovery from sensory disturbance after sagittal splitting ramus osteotomy using a Semmes-Weinstein pressure aesthesiometer.

A number of studies of evaluation methods for sensory disturbance after sagittal splitting ramus osteotomy (SSRO) are known. To compare postoperative sensory disturbances among patients in different hospitals, a highly reproducible and standardized sensory test is required. In the present study, we measured the tactile threshold in the region innervated by the mental nerves in 45 patients (90 sides) after SSRO using a Semmes-Weinstein pressure aesthesiometer. The percentage of recovery to the normal level defined by Bell was 72.2% at one week, 82.2% at 4 weeks, and 90.0% at 8 weeks after SSRO. The recovery process was evaluated by classifying the postoperative sensory disturbance into 5 grade levels according to Bell's interpretation scale. The results showed that the SW sensory test is useful for evaluation of the recovery process from sensory disturbance after SSRO. Some improvements of this test were also discussed.

A case of desmoplastic ameloblastoma of the maxilla.

A case of desmoplastic ameloblastoma of the maxilla is presented. The tumor showed characteristic radiographical and histological features. Radiographically, the tumor showed a mixed radiolucent-radiopaque lesion occupying the left maxillary sinus with undefined borders in the alveolar process. Histopathologically, the tumor was characterized by abundant, densely collagenous stroma with small nests and strands of odontogenic epithelium and bone tissues. Findings during operation suggested that the tumor arising in the alveolar process had grown upwards and almost displaced the left maxillary sinus. Complete resection is recommended for this variant because of the possibility of tumor infiltration into the surrounding bone marrow spaces.

Use of the buccal fat pad as a pedicle graft.

A technique was developed to use the pedicle buccal fat pad as method for closing an oro-antral communication. The buccal fat pad is situated in the posterior part of the maxilla and bounded by the pterygopalatine fossa, buccal muscle, and masseter muscle. It is supplied by blood from small branches of the maxillary artery, the superficial temporal artery, and the facial artery. Generally, the buccal fat pad can be easily reached by a horizontal incision over the periosteum near the maxillary third molar. As much as 60 x 50mm2 can be obtained. For reconstructive surgery, the buccal fat pad is suitable for closing the sinus near the hard and soft palate and naso-oral fistulae. In our case, graft tissues after surgery healed satisfactorily, began to epithelialize at two weeks after operation, and appeared with normal mucous-like changes at about one month. Thus, the pedicle buccal fat pad is a safe, simple, convenient, and effective grafting method, if it is applied to appropriate cases.

Transposition of the mental nerve and inferior alveolar nerve trunk.

Transposition of the mental nerve trunk is performed as a preprosthetic procedure. It is effective for patients with hyperesthesia caused by dental prosthesis and resorption of the alveolar ridge caused by aging. It is also performed to protect a location for implant insertion and to preserve the inferior alveolar nerve during mandibular tumor excision surgery. The demand for this surgery is increasing. The disadvantages of the surgery performed with the usual methods include the difficulty of the procedure and the long operation time. In an attempt to solve these problems, this study was conducted at the Department of Oral and Maxillofacial Surgery, Medical University Hanover, introducing an improved surgical method and presenting the actual operation performed in our department. This surgical method is considered superior because it is easy to perform, requires no special equipment and results in recovery of the mental nerve after surgery.

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.

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.

Iontophoretic delivery of amino acids and amino acid derivatives across the skin in vitro.

The effects of penetrant properties (lipophilicity and charge) and of vehicle pH on the iontophoretically enhanced delivery of amino acids and their N-acetylated derivatives have been examined in vitro. The penetrants were nine amino acids (five were zwitterionic, two positively charged, and two negatively charged) and four N-acetylated amino acids, which carry a net negative charge at pH 7.4. Iontophoresis at constant current (0.36 mA/cm2), using Ag/AgCl electrodes, was conducted across freshly excised hairless mouse skin. Iontophoretic flux of the zwitterions was significantly greater than passive transport. Delivery from the anode was greater than from the cathode for all zwitterions. The level of enhancement was inversely proportional to permeant octanol/pH 7.4 buffer distribution coefficient. Cathodal iontophoresis of the negatively charged amino acids and of the N-acetylated derivatives produced degrees of enhancement which were significantly greater than those measured for the "neutral" zwitterions. Furthermore, the enhanced flux reached a steady-state level within a few hours for the negatively charged species, whereas the transport of the zwitterions continued to increase with time. Anodal iontophoresis of histidine and lysine, the two positively charged amino acids studied, induced substantial enhancement which was sensitive to the pH of the delivery vehicle. For example, the flux of histidine from an applied solution at pH 4 (where the amino acid carries a net positive charge) was significantly greater than that from a vehicle at pH 7.4 (where histidine is essentially neutral). The behavior of lysine was more complex and suggested a certain degree of neutralization of the skin's net negative charge.

Ultrastructural study of the histogenesis of salivary gland mucoepidermoid carcinoma.

This electronmicroscope study of the histogenesis of mucoepidermoid carcinoma is based on 15 cases from the major and minor salivary glands. Six major cell types were identified: undifferentiated (cuboid) stem cells, intermediate (columnar) cells, serous/mucoid secretory cells, mucus-producing goblet cells, squamous/epidermoid cells and myoepithelium. It is proposed that undifferentiated stem cells serve as pluripotential reserves which give rise to the various cell types seen in mucoepidermoid carcinoma. Reserve cells in the acinar-intercalated duct components of the salivary glands appeared to give rise to the serous/mucoid and myoepithelial cell populations of mucoepidermoid carcinoma. Reserve cells in the proximal ducts system (intra/extralobular striated ducts and excretory ducts) differentiated into myoepithelium, intermediate (columnar) cells, squamous/epidermoid and mucus-producing goblet cell lines. It is proposed that neoplastic reserve cells give rise to different tumor types in the salivary glands. These types result from varying admixtures and arrangements of tumor cells at different stages of their structural and functional cytodifferentiation from reserve cells. It is further proposed that neoplastic reserve cells differentiate along similar cell lines as the embryonic "stem cells" in the development of normal salivary glands.

Ultrastructure of normal human parotid gland with special emphasis on myoepithelial distribution.

The myoepithelium in human parotid, unlike that in other species, is not limited to the acinar-intercalated ductal system but extends to the intra- and extralobular striated ducts. Myoepithelial cells of the striated ducts are smaller, fewer, and different in shape but they are similar in their relationships and ultrastructural morphology to the acinar-intercalated system. Distribution of myoepithelium is discussed with respect to its role and participation in the histogenesis of salivary gland tumours, especially those of intermediate and large duct cell origin.