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Homeobox genes play important roles in craniofacial morphogenesis. However, the characteristics of the transcription factor Hoxc during palate formation remain unclear. We examined the immunolocalization patterns of Hoxc5, Hoxc4, and Hoxc6 in palatogenesis of cleft palate (Eh/Eh) mice. On the other hand, mutations in the FGF/FGFR pathway are exclusively associated with syndromic forms of cleft palate. We also examined the immunolocalization of Fgfr1 and Erk1/2 to clarify their relationships with Hoxc in palatogenesis. Some palatal epithelial cells showed Hoxc5 labeling, while almost no labeling of mesenchymal cells was observed in +/+ mice. As palate formation progressed in +/+ mice, Hoxc5, Hoxc4, and Hoxc6 were observed in medial epithelial seam cells. Hoxc5 and Hoxc6 were detected in the oral epithelium. The palatal mesenchyme also showed intense staining for Fgfr1 and Erk1/2 with progression of palate formation. In contrast, the palatal shelves of Eh/Eh mice exhibited impaired horizontal growth and failed to fuse, resulting in a cleft. Hoxc5 was observed in a few epithelial cells and diffusely in the mesenchyme of Eh/Eh palatal shelves. No or little labeling of Fgfr1 and Erk1/2 was detected in the cleft palate of Eh/Eh mice. These findings suggest that Hoxc genes are involved in palatogenesis. Furthermore, there may be the differences in the localization pattern between Hoxc5, Hoxc4, and Hoxc6. Additionally, Hoxc distribution in palatal cells during palate development may be correlated with FGF signaling. (228/250 words) © 2016 Japanese Teratology Society.

2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) induces cleft palate and hydronephrosis in the mouse embryo. Cleft palate occurs due to failure in palatal grow, but the underlying mechanisms are unclear. We investigated the mechanisms of cleft palate development in TCDD-exposed mouse embryos. We administered olive oil (control group) or TCDD diluted in olive oil (40 μg/kg) via gastric tubes to pregnant mice on gestational day (GD) 12. Embryos of control and TCDD-exposed groups were removed from pregnant mice on GD 14 and GD 15, respectively. One mouse embryo from the control group had anteroposterior palatal fusion. Palatal fusion was observed in three TCDD-exposed mouse embryos. Palates of TCDD-exposed mice fused from the interior to the middle of the palates, while the palates were separated in the posterior region. The middle of the embryonic palatal shelves in TCDD-exposed animals was narrow and split at the fusional position. At this position, palatal and blood cells were dispersed from the palatal tissue and the epithelium was split, with a discontinuous basement membrane. The results suggest that decreased intercellular adhesion or insufficient tissue strength of the palatal shelves may be involved in the development of cleft palate following palatal fusion.

Mouse embryos exposed to 2,3,7,8-tetrachloridedibenzo-p-dioxin (TCDD) develop cleft palates and hydronephrosis. Cleft palates occur after TCDD exposure due to contact and/or fusion failure. We investigated whether cleft palate can be induced by dissociation of the palatine process after fusion. Pregnant mice on gestational day (GD) 12 were randomly divided into two groups: one group was administered through gastric tubes one dose of olive oil (control group) and the other group was administered one dose of TCDD diluted with olive oil, both at a dose of 40 microg/kg body weight. Embryos were removed by cesarean section from pregnant mice during the palatal formation stage (GD 13-18) and the palatal form was observed using a stereoscopic microscope. In TCDD-exposed embryos, palatal fusion was observed on GD 14, 15 and 16 and the incidence of cleft palate was 100% on GD 18. Fusion rates were 17.5 +/- 15.2% and 12.4 +/- 11.8% on GD 15 and 16, respectively. Some palates from the TCDD-exposed mouse embryos showed clearly developed cleft palate after fusion of the lateral palatine processes during palatal formation. A mass of cells, which were chiefly epithelial in the fused palates was observed in the TCDD-exposed mouse embryos. A decrease in E-cadherin expression was observed in this mass of cells, indicating its involvement in the development of cleft palate.

A case of infantile malignant osteopetrosis with refractory mandibular osteomyelitis is reported. A female child was diagnosed with osteopetrosis at 2 years 8 months of age, and was scheduled to receive a bone marrow transplantation (BMT) in a pediatric department at 3 years 11 months of age. Her lower incisors were extracted at a dental clinic, after which she had recurring abscesses with progressive severity. She was referred to our department to control local infections before the BMT and was diagnosed with chronic mandibular osteomyelitis caused by osteopetrosis. Sequestrotomy and curettage were performed under general anesthesia. After surgery, daily saline irrigation was continued and osteomyelitis was well controlled. An uneventful BMT was performed, although it was refused and failed. A second BMT was planned, but during chemotherapy, cellulitis occurred after a recurrence of osteomyelitis caused by a newly erupted tooth. The local infection was controlled, but pneumonia recurred. She ultimately died of respiratory failure at 5 years of age.

Summary Objective techniques to evaluate a facial movement are indispensable for the contemporary treatment of patients with motor disorders such as facial paralysis, cleft lip, postoperative head and neck cancer, and so on. Recently, computer-assisted, video-based techniques have been devised and reported as measuring systems in which facial movements can be evaluated quantitatively. Commercially available motion analysis systems, in which a stereo-measuring technique with multiple cameras and markers to facilitate search of matching among images through all cameras, also are utilized, and are used in many measuring systems such as video-based systems. The key is how the problems of facial movement can be extracted precisely, and how useful information for the diagnosis and decision-making process can be derived from analyses of facial movement. Therefore, it is important to discuss which facial animations should be examined, and whether fixation of the head and markers attached to the face can hamper natural facial movement.

This case report describes the treatment of a patient with bimaxillary protrusion and masseter muscle hypertrophy. At age 21 years 7 months, this woman had temporomandibular disorder (TMD) symptoms, severe bimaxillary protrusion, and a prominent mandibular angle with facial asymmetry. After an attempt to alleviate the TMD symptoms with occlusal splint stabilization, portions of the masseter muscle and the mandible were surgically removed. Titanium screws were placed bilaterally in both arches, and a retraction force was applied. After active treatment for 38 months, the convexity of the facial profile with lip protrusion was improved remarkably, and good occlusion was achieved. The prominent mandibular angle with facial asymmetry was improved as a result of the surgical reduction of the masseter muscle and the modeling ostectomy near the masseteric tuberosity. The TMD symptoms disappeared, and the jaw movement pattern became normal. Therefore, our results suggest that this combination treatment would be useful for masseter muscle hypertrophy for morphologic and functional problems.

Objective: To assess the usefulness of maxillo-mandibular fixation (MMF), occlusal and skeletal stabilities were evaluated after bilateral sagittal splitting ramus osteotomy (BSSRO) with or without MMF to treat mandibular prognathism or asymmetry.Patients and Methods: Thirty-three cases who underwent surgical correction of mandibular prognathism or asymmetry were analyzed. The surgical procedure in all cases consisted of SSRO. The segments were fixed rigidly with titanium screws. A postoperative MMF was performed in 16 patients within one day of the SSRO. The MMF was placed for one week. The other 17 patients were free to move their jaw on the day of the surgery, and received occlusal guidance with elastics starting from the third postoperative day. Cephalograms were taken preoperatively, at 1 day postoperatively, and at 1, 3, 6, 12 and 24 months after surgery. Skeletal and occlusal stabilities were then assessed.Results: Even without MMF, occlusion was guided to the planned position with soft elastics after surgery. Later than 1 month, occlusal and skeletal stability were satisfactory in both groups, and there was no correlation between the surgical results and post-operative MMF.Conclusion: MMF may not be necessary after a rigid fixation SSRO to treat mandibular prognathism or asymmetry, considering the risks of airway problems.

Cementogenesis starts with the differentiation of cementoblasts. Mature cementoblasts secrete cementum matrix. Cementum components are similar to bone; moreover, cementoblasts possess many characteristics similar to those of osteoblasts. Runx2 and osterix, the transcriptional factors for osteoblast differentiation, participate in tooth formation. However, the characteristics of Runx2 and osterix during the differentiation process of cementoblasts remain unclear. In this study, we examined the immunolocalization patterns of Runx2, osterix, and osteopontin during rat molar tooth formation. Periodontal ligament cells and osteoblasts located on the alveolar bone surface showed immunoreactivity for Runx2. Colocalization of Runx2 and osterix was detected in cementoblasts, which penetrated the ruptured Hertwig's epithelial root sheath and attached to root dentin. Moreover, osteopontin was observed in Runx2-positive cementoblasts facing the root surface. However, the cells adjacent to cementoblasts showed only Runx2 reactivity. Neither Runx2 nor osterix was seen in cementocytes. These results suggest that both Runx2 and osterix are important for differentiation into cementoblasts. Additionally, osterix may be indispensable for transcription of osteopontin expression. (J Histochem Cytochem 57:397–403, 2009)

The aim of this study was to assess the necessity of maxillo-mandibular fixation (MMF) from a viewpoint of postoperative stability to treat mandibular asymmetries. Twenty-four patients who underwent surgical correction of mandibular asymmetry were analyzed. The surgical procedure in all patients consisted of bilateral sagittal splitting ramus osteotomy (SSRO). The segments were then fixed rigidly with titanium screws. A postoperative MMF was performed in 12 patients within 1 day of the SSRO. The MMF lasted for 1 week. The other 12 patients were free to move their jaw on the day of the surgery and received occlusal guidance with elastics starting from the third postoperative day. Posterior–anterior cephalograms were taken preoperatively, 1 day postoperatively, and at 1, 3, 6, 12, and 24 months after surgery. Skeletal and occlusal stabilities along with postoperative complications were then assessed. Nausea and pharyngeal discomforts were observed very often in both groups. Even without MMF, occlusions were guided to the objective positions by an average of 3.5 days after surgery. Occlusal and skeletal stability was satisfactory in both groups, and there was no correlation between the surgical results and the use of postoperative MMF. MMF is not necessary after rigid fixation SSRO for mandibular asymmetry, considering the risks of airway distress.

Objectives and design: The expressions of human beta defensin-1 (HBD-1), -2 (HBD-2) and -3 (HBD-3) in non-inflamed pseudocysts such as mucoceles were investigated immunohistochemically in this study. Materials and methods: Mucocele specimens were obtained from 21 patients. The expression of HBDs was studied immunohistochemically by using antibodies directed against HBD-1, -2, and -3. Statistical analyses were carried out on serial sections stained with antibodies. Results: Cells expressing HBDs were found in mucoceles. The expression of HBD-2 was observed in floating cells in all the specimens, whereas HBD-1 and HBD-3-expressing cells were detected in 93% and 73% of the mucoceles, respectively. The HBD-2 signal was the most intense and the HBD-3 signal intensity was weaker than that of HBD-1. HBDs were expressed in neutrophils and in other floating cells. Interestingly, the signal intensity and the population of positive cells located close to the centers of cysts were higher than those located in the peripheral areas of cysts. Conclusion: The expression of HBDs was found even in non-inflamed pseudocysts such as mucoceles. These results suggest that an unknown mechanism not involved in biophylaxis for the expression of HBDs may exist.

The objective of this study was to assess the usefulness of maxillo-mandibular fixation (MMF) from a viewpoint of skeletal and occlusal stability and to investigate the complications after surgery with and without MMF.Twenty-seven patients who underwent surgical correction of class III malocclusion were analyzed. The surgical procedure in all cases consisted of a bilateral sagittal splitting ramus osteotomy (SSRO). The segments were then fixed rigidly with titanium screws. A postoperative MMF was performed in 13 patients within 1 day of the SSRO. The MMF lasted for 1 week. The other 14 patients were free to move their jaw on the day of the surgery and received occlusal guidance with elastics starting from the third postoperative day. Cephalograms were taken preoperatively, at 1 day postoperatively and at 1, 3, 6, 12, and 24 months after surgery. Skeletal and occlusal stabilities along with postoperative complications were then assessed.Discomfort of the pharynx and postoperative pain was more severe in the MMF group. Even without MMF, occlusion was guided to the ideal position by an average of 5.3 days after surgery. Occlusal and skeletal stability was satisfactory in both groups, and there was no correlation between the surgical results and postoperative MMF.MMF may not be necessary after a rigid fixation SSRO, considering the risks of airway problems.

Morphological and immunohistological examinations were performed to reveal the mechanisms of cleft palate induction by 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). ICR strain mice 8-10 weeks of age were used in the study. TCDD was administered in olive oil on gestation day (GD) 12.5 with gastric tubes at 40 microg/kg. From GD 13.5 to 16.5, palates were examined by scanning electron microscopy (SEM), hematoxyline-eosin (HE) staining, and immunohistochemical staining of FGFR1/2, TGF-beta3, MSX1 and LHX8. In the control group, both of the palatal shelves began elevating on GD 14.0 and finished within 6 h. After the elevation, all of the shelves had completely fused with each other on GD 14.5. In the TCDD-treated group, palatal shelves elevated 1 day later than in the control group. However, all palates had elevated by GD 15.0. After the elevation, the shelves contacted each other and fused; however, they were separated on GD16.0. HE staining showed that medial edge epithelium (MEE) was thinner in the TCDD group than in the control group. MEE observed under a high magnification (x2500) exhibited filopodia-like filaments and the cells were bulged in the control group. In contrast, in the TCDD group, no filaments were observed and the cells were flat with unclear boundaries. Immunohistologically, there were no characteristic findings except for FGFR1. FGFR1 was not expressed in the TCDD group after the fusion phase (GD 14.5). TCDD induces many morphological and molecular changes to MEE cells and causes cleft palates.

Both periosteum and bone marrow have the potential to induce heterotopic bone when grafted. Whether the process of bone formation is controlled by the recipient environment where the donor graft is placed or by factors from the donor site is not well documented. The purpose of this study was to examine the histology of new bone induced by either autogenously grafted periosteum or autogenously grafted bone marrow using the rat calvarial defect model in Sprague-Dawley rats. Grafts of either bone marrow or periosteum obtained from tibias were placed in calvarial defects with beta-tricalcium phosphate. Ten days after grafting, active cell proliferation was observed in the defects of both types of grafts. After 20 days, cancellous bone formation was observed in the defects with bone marrow grafts, and intramembranous bone formation was observed in the defects with periosteal grafts. After 30 days, bone marrow grafts had developed bone with a bone marrow-like structure, and the periosteal grafts had produced cortical bone structure in the defects. The findings suggest that the type of bone formation is determined by characteristics of the donor site.

Objective: To investigate respiratory status and other clinical conditions pre- and postoperatively following sagittal splitting ramus osteotomy setback surgery. Patients and Methods: Eleven patients who underwent surgical correction of class III malocclusion were analysed. The surgical procedure consisted of bilateral sagittal splitting ramus osteotomy. The segments were fixed rigidly with titanium screws. For preservation of the airway, maxillomandibular fixation was not performed. The patients' respiratory status was evaluated using an apnoea monitor preoperatively, on the day of operation, 1 week postoperatively, and at more than 6 months postoperatively. The patients' pharyngeal morphology was also assessed by lateral cephalograms preoperatively, and 1 day and 6 months postoperatively. Results: The posterior airway space was decreased significantly immediately after surgery, but no remarkable changes were observed in the following 6 months. The Apnoea Hypopnoea Index and lowest saturation pulse oximetry were generally unaffected. However, in some patients, the respiratory status became worse immediately after surgery. Conclusions: Obstructive sleep apnoea syndrome is usually not provoked by mandibular setback surgery by the sagittal splitting ramus osteotomy procedure, despite the associated pharyngeal narrowing. However, careful observation is necessary because obstructive sleep apnoea syndrome is possible following surgery.

Pleomorphic adenoma (PA) is the most common salivary gland tumor. Tumors originating from the major salivary glands are frequently located in the parotid gland. In contrast, tumors originating from the minor salivary glands are frequently found in the palatal region. PA occurring in the lips, especially the lower lip, is relatively rare.In this paper, we present 2 cases of PA arising in the upper lip and lower lip, respectively.The first case arose in a 72-year-old woman. The patient visited a dental clinic because of swelling of the left side of the upper lip and was referred to our department. Palpation revealed a painless movable mass. The lesion was 15 mm in diameter and located in the subcutaneous tissue on the left side of the upper lip.The second case was in a 60-year-old man. This patient visited a dental clinic because of swelling of the left side of the lower lip and was referred to our department. Palpation revealed a painless movable mass. The lesion was 13 mm in diameter and arose in the subcutaneous tissue of the left side of the lower lip.The clinical diagnosis in both cases was a benign tumor of the lip. Surgical resection was performed under local anesthesia in both patients, and the histopathological diagnoses confirmed PA. In both patients, the upper and lower lips healed well after the operation. About 1 and a half years after surgery, there was no evidence of recurrence.

Objective : 2,3,7,8-Tetrachlorodibenzo- p -dioxin (tetrachlorodibenzodioxin) is known to cause cleft palates in pregnant mice in vivo. This study aimed at investigating the effects of tetrachlorodibenzodioxin on murine palatogenesis in vitro using organ culture. Materials and Methods : Palatal shelves from ICR mice foetuses (gestational day, 12.5) were removed and dissected to isolate the palatal tissue. Tissues were cultured in a chemically defined, serumless medium using a suspension culture technique. Palatal fusion rates were assessed in suspension cultures containing tetrachlorodibenzodioxin at concentrations of 0.1, 1, 10 and 50 ng/mL and in non-treated controls. Palates from the control and 10 ng/mL groups were stained immunohistologically with antibodies against cleft palate-related factors (transforming growth factor-beta3, muscle segment homeobox 1, Lim-homeobox 8, fibroblast growth factor receptor 1, fibroblast growth factor receptor 2). Results : Over 90% of the palatal shelves completely fused in suspension culture after careful removal of the neural tissues. With tetrachlorodibenzodioxin addition, the palatal fusion rate decreased to 50% at a concentration of 1 ng/mL, however, the rate did not decrease any further at higher concentrations. Immunohistologically, transforming growth factor-beta3 disappeared earlier in the tetrachlorodibenzodioxin group. Conclusions : Tetrachlorodibenzodioxin contributes to the pathogenesis of cleftpalate in vitro; however, direct intracellular effects are insufficient to fully accountfor the observed effects of tetrachlorodibenzodioxin, and it is possible that indirect mechanisms are also in play in vivo.

Objective: Activating mutations within the stimulatory G protein alpha gene are typically associated with the McCune-Albright Syndrome. These mutations have been found in endocrine tumours, intramuscular myxomas, and polyostotic fibrous dysplasias of bone in patients with McCune-Albright syndrome. We hypothesised that the same mutations might occur in odontogenic myxoma and monostotic fibrous dysplasia of bone in the jaw. In this study, we aimed to clarify the association between these tumours and mutations in the stimulatory G protein alpha gene. Patients and Methods: Paraffin-embedded specimens of 5 cases each of odontogenic myxoma and monostotic fibrous dysplasia of bone in the jaw were examined. Exon 8 and exon 9 of the stimulatory G protein alpha gene were screened by polymerase chain reaction-single-stranded conformation polymorphism analysis, and the polymerase chain reaction products were then sequenced using the dideoxy method. Result: No activating mutations of the stimulatory G protein alpha gene were found in any of the cases of odontogenic myxoma or monostotic fibrous dysplasia of bone in the jaw. Conclusion: Odontogenic myxoma and monostotic fibrous dysplasia of bone in the jaw are most likely not associated with activating mutations of the stimulatory G protein alpha gene.

Periosteum is a source of osteoprogenitor cells and some investigators advocated the effective use of periosteum as a grafting material for the repair of bone and joint defects. In the present study, we showed bone formation induced from cultured periosteum-derived cells (CPDC) in the rat calvarial defect model. Periosteum taken from a rat tibia was immediately placed in culture medium with 10% fetal bovine serum. After confluence, periosteal cells were re-cultured three dimensionally on a collagen sponge for 7 days. Periosteal cell/collagen complex was grafted in calvarial defect of rats. At 30 and 60 days post grafting, grafted tissue was extracted and compared histologically and radiographically with control groups. In the experimental group at 30 days after implantation, new bone formation was seen. Radiographically, mineralized new bone formation was revealed in the defect at 60 days post grafting. New bone formation in periosteal cell/ collagen complex was greater than the other groups. Our results suggest that CPDC may have an important role in bone formation in the calvarial defect in rats.

The purpose of this study was to evaluate the clinical application of endosseous implants for management of grafted alveolar cleft. Four patients (1 male and 3 female; mean age 26.6 years; age ranging from 21.5 to 30.4 years at first implant surgery) with unilateral cleft lip and palate were evaluated. All patients received autogenous particulate cancellous bone and marrow graft (PCBM) to close the alveolar cleft. After bone bridge formation, implant treatment was performed. All patients with insufficient alveolar bone height, chin or ramus of mandible bone, onlay grafting was performed 4 months before implant installment. A total of 5 implants were placed in the grafted alveoli. The length of implants ranged from 10.0 to 13.0 mm. The follow-up period ranged from 2.2 to 4.1 years after implant placement. The clinical result was acceptable in all patients. The survival rate of implants was 100%. Implant treatment for grafted alveolar cleft was studied and the outcome of treatment was acceptable.

In the oral cavity, epidermoid cysts usually arise in the oral floor and rarely occur in the upper lip. A case of an epidermoid cyst of the upper lip is reported. The patient was a 22-year-old man. A cyst arising in the left side of the upper lip was diagnosed by the patients former doctor, and the lesion was extirpated under local anesthesia 1 year 7 months ago. However, 6 months ago, a painless mass developed slowly on the left side of the upper lip. Palpation, revealed a large movable mass. MRI and ultrasonic examinations showed that the lesion was 25mm in diameter in the subcutaneous tissue of the left side of the upper lip. The lesion was round and homogenous inside. We diagnosed the lesion to be a cyst of the upper lip and extirpated the mass under general anesthesia. The excised material was 20×10 mm in diameter and filled with a yellowish white serous liquid and a yellowish white cheese-like material. The specimen was histopathologically diagnosed to be an epidermoid cyst. Two years after the operation, the left side of the upper lip has healed and there is no evidence of recurrence.

Homeobox genes play important roles in craniofacial morphogenesis. However, the characteristics of the transcription factor Hoxc during palate formation remain unclear. We examined the immunolocalization patterns of Hoxc5, Hoxc4, and Hoxc6 in palatogenesis of cleft palate (Eh/Eh) mice. On the other hand, mutations in the FGF/FGFR pathway are exclusively associated with syndromic forms of cleft palate. We also examined the immunolocalization of Fgfr1 and Erk1/2 to clarify their relationships with Hoxc in palatogenesis. Some palatal epithelial cells showed Hoxc5 labeling, while almost no labeling of mesenchymal cells was observed in +/+ mice. As palate formation progressed in +/+ mice, Hoxc5, Hoxc4, and Hoxc6 were observed in medial epithelial seam cells. Hoxc5 and Hoxc6 were detected in the oral epithelium. The palatal mesenchyme also showed intense staining for Fgfr1 and Erk1/2 with progression of palate formation. In contrast, the palatal shelves of Eh/Eh mice exhibited impaired horizontal growth and failed to fuse, resulting in a cleft. Hoxc5 was observed in a few epithelial cells and diffusely in the mesenchyme of Eh/Eh palatal shelves. No or little labeling of Fgfr1 and Erk1/2 was detected in the cleft palate of Eh/Eh mice. These findings suggest that Hoxc genes are involved in palatogenesis. Furthermore, there may be the differences in the localization pattern between Hoxc5, Hoxc4, and Hoxc6. Additionally, Hoxc distribution in palatal cells during palate development may be correlated with FGF signaling. (228/250 words) © 2016 Japanese Teratology Society.

Objective: To characterise a natural antisense transcript of connective tissue growth factor. Materials and Methods: RNA from several cell lines was analysed by RNase protection assay to detect antisense transcripts. To characterise the regulatory aspect of the transcribed area, chimeras were constructed in which the firefly luciferase gene was fused with the corresponding segment of ctgf/ccn2, and the gene expression was monitored. Results: A natural antisense transcript complementary to the 3'-untranslated region of ctgf/ccn2 mRNA in cultured human tumour cells was detected. The luciferase gene fused with the full-length antisense 3'-untranslated region showed strikingly low levels of luciferase expression compared with the control. Based on a series of deletion analyses, the major repressive element was located in the middle portion within the 3'-untranslated region, which corresponded to the antisense transcribed area. Conclusion: These results suggest that controlled expression of antisense RNAs against the 3'-untranslated region of ctgf/ccn2 mRNA may be involved in the determination of phenotypes in certain oral malignancies.

Microcystic adnexal carcinoma is a rare tumour, first described in 1982. Prior to the recognition of microcystic adnexal carcinoma, instances of unusual basal cell carcinoma, sweat gland carcinoma, and trichofolliculoma had been reported. Microcystic adnexal carcinomas are slow growing and demonstrate aggressive local behaviour and a high recurrence rate. The mainstay of treatment is surgical excision using frozen sections to check the peripheral and deep margins. This report is of a microcystic adnexal carcinoma in the mental region of a 69-year-old woman.

Objective: To investigate the possible involvement of 2,3,7,8-tetrachlorodibenzo-p-dioxin in the pathogenesis of cleft palate in mouse palatal cells. Materials and Methods: Palatal shelves of gestation day 12.5 ICR mouse foetuses were removed and cultured with serum. One day after the second passage, the medium was changed to serum free and 2,3,7,8-tetrachlorodibenzo-p-dioxin was added at 1 ng/mL, 10 ng/mL, and 100 ng/mL. After culture for 1 and 3 hours, all cellular RNA was isolated from the cells. Expression of some cleft palate-related genes was analysed semiquantitatively using reverse transcriptase-polymerase chain reaction. Five genes (Tgfb3, Msx1, Lhx8, Fgfrl, and Fgfr2) were selected on the basis that functional disturbance of these genes causes cleft palate with high frequency. The data were compared with GAPDH. Results: In the first passage, many mouse embryonic palatal mesenchymal cells had spread out, and a number of monolayer spindle-shaped cells were identified. The cells were stable in the second passage and also in the serum free medium. One hour after addition of 2,3,7,8-tetrachlorodibenzo-p-dioxin, Tgfb3 mRNA expression was reduced at a concentration of 100 ng/mL. After 3 hours, Tgfb3, Msxl, and Lhx8 mRNA expression were also reduced. However, there was no effect of 2,3,7,8-tetrachlorodibenzo-p-dioxin on the gene expression for Fgfr1 and Fgfr2 mRNA. Conclusion: 2,3,7,8-Tetrachlorodibenzo-p-dioxin may contribute to the induction of cleftpalate by suppressing Tgfb3, Msx1, and Lhx8 gene expression.

Objective : To investigate the expression of Sox9 protein in the process of cartilage formation after periosteal grafting using the rat periosteum graft model. Materials and Methods : Periostea taken from twenty 7-week-old rats were immediately grafted into the suprahyoid muscles. Grafted tissues were harvested 0, 7, 14, and 21 days after grafting and examined by microscopy and immunohistochemistry using CD44, type 2 collagen, and Sox9 antibodies as markers. Results : Periosteal cells expressed CD44 after 7 days and Sox9 after 14 days. The Sox9-positive cells showed chondrogenic differentiation with type 2 collagen expression by 21 days after grafting. Endochondral ossification was also seen 21 days after grafting. Sox9 was not detected in hypertrophic chondrocytes. Conclusion : These findings suggested that Sox9 may be related to chondrogenic differentiation of periosteal cells in grafted periosteum.

Connective tissue growth factor (CTGF/CCN2) is a potent angiogenic factor. In this report, we describe for the first time that vascular endothelial growth factor (VEGF)–mediated induction of the ctgf/ccn2 gene was a post-transcriptional event that was inhibited by a novel angiogenic inhibitor, DN-9693, in human umbilical vein endothelial cells. Steady-state mRNA levels of ctgf/ccn2 were remarkably increased by VEGF in a concentration-dependent manner, whereas the activity of the ctgf/ccn2 promoter was not responsive to VEGF as confirmed by a reporter gene assay and quantitative real-time PCR analysis. By employing a RNA degradation assay, we eventually found that the observed increase in the ctgf/ccn2 mRNA level was due to an increased stability of the mRNA induced by VEGF. DN-9693 at a dose of 0.1 to 2 ng/mL did not affect basal levels of ctgf/ccn2 mRNA; however, enhancement of ctgf/ccn2 mRNA expression by VEGF was specifically inhibited by DN-9693. Of importance, the inhibitory effects could be also ascribed to post-transcriptional regulation, because the VEGF-mediated increase in stability of ctgf/ccn2 mRNA was suppressed by DN-9693. Furthermore, we investigated the effects of DN-9693 on VEGF-induced activation of three subgroups of mitogen-activated protein kinase pathways and found that DN-9693 blocked the activation of these pathways by VEGF. These results suggest that VEGF increases ctgf/ccn2 mRNA stability through mitogen-activated protein kinase–mediated intracellular signaling cascade(s), which can be inhibited posttranscriptionally by a novel angiogenic inhibitor, DN-9693, in human umbilical vein endothelial cells. [Mol Cancer Ther 2006;5(1):129–37]

Objective: To introduce a new method for obtaining 3-dimensional facial images and evaluating soft tissue changes after surgery. Patients and Methods: Six orthognathic surgery patients who underwent mandibular setback procedures by bilateral sagittal split ramus osteotomy were evaluated. The movements performed during surgery were estimated by lateral cephalogram tracings. The soft tissue changes were evaluated by the use of a noncontact 3-dimensional laser scanner Changes were analysed by measuring the distance of the normal vectors from the preoperative scan surface to the surface of the postoperative scan at each pixel. The shape change pattern was analysed at the mental and inferior lip areas. Results: The amount of mandibular retrusion ranged from 4.2 mm to 17.1 mm at the bony pogonion. The 3-dimensional scans showed that all patients presented a similar change pattern in which the greatest amount of soft tissue change was close to the midline and gradually diminished laterally. Conclusions: The scanning method used in this study to obtain the facial scan was simple, fast, and reliable. Since the direction of the normal vectors analysed was dependent on the facial surface, change was analysed in multiple directions and represented by a coloured facial map. Despite the statistical limitations of this system, it provides the clinician with a visual and comprehensible image to represent shape changes.

Connective tissue growth factor (CTGF/CCN2) can be induced by various forms of stress such as exposure to high glucose, mechanical load, or hypoxia. Here, we investigated the molecular mechanism involved in the induction of ctgf/ccn2 by hypoxia in a human chondrosarcoma cell line, HCS-2/8. Hypoxia increased the ctgf/ccn2 mRNA level by altering the 3'-untranslated region (UTR)-mediated mRNA stability without requiring de novo protein synthesis. After a series of extensive analyses, we eventually found that the cis-repressive element of 84 bases within the 3'-UTR specifically bound to a cytoplasmic/nuclear protein. By conducting a UV crosslinking assay, we found the cytoplasmic/nuclear protein to be a 35 kDa molecule that bound to the cis-element in a hypoxia-inducible manner. These results suggest that a cis-element in the 3'-UTR of ctgf/ccn2 mRNA and trans-factor counterpart(s) play an important role in the post-transcriptional regulation by determining the stability of ctgf/ccn2 mRNA.

Objective: To investigate the effect of naso-labial scarring on the skeletal growth pattern of the maxillary complex. Materials and Methods: Eighty eight naturally delivered ICR-strain infant mice were divided into 5 groups based on the site of the scar Each group was further divided into 3 subgroups according to the age of the animal at the time of surgery (1 day, 1 week, or 3 weeks). In group 1, the left upper lip was incised from the nostril floor to the vestibule using electrocautery. In group 2, the left upper lip and alveolus were incised from the nostril floor to the primary palate using scissors. In group 3, the nasal septum was released through the vestibule using a chisel. In group 4, the upper vestibule between the angles of the mouth was incised using electrocautery. In group 5, no wound was made in age-matched mice to act as a control group. The animals were weighed and killed at the age of 7 weeks, and standardised dorsoventral cephalograms were taken. Results: The scars of the lip and alveolus caused a decrease in the width of the anterior part of the maxilla, and surgery at 1 week caused a decrease in the anteroposterzor length of the maxilla. Surgery to the nasal septum had a severe influence on growth. Surgery to the vestibule caused a decrease only in the maxillary length. Conclusion: Although the results cannot be extrapolated to humans, the study serves to flag the vestibular and septal surgery elements of cleft lip repair for further evaluation.

Ameloblastoma is the most common odontogenic neoplasm, particularized by its local invasiveness. Heparanase is the endo-glucuronidase enzyme that specifically cleaves heparan sulfate, the important modulator of extracellular matrix, and related to invasion of tumor cells. In this study, we addressed to show the gene expression and localization of heparanase in ameloblastoma. Immunohistochemistry and in situ hybridization of heparanase were carried out in 23 ameloblastomas. Strong expression of heparanase at both mRNA and protein levels was detected in all ameloblastomas studied. Small tumor nests and budding epithelial branches showed stronger staining pattern and the stromal tissues at the immediate vicinity of the tumor nests with strong heparanase expression were loose and edematous. Cystic areas and squamous metaplastic areas of the tumor showed intense staining with heparanase antibody proposing the implication of heparanase in these processes. These results suggest the possible contribution of heparanase in the local invasiveness and secondary morphologic changes of ameloblastoma.

Background: Distraction osteogenesis (DO) has become a mainstream surgical technique for patients with jaw deformities. In cases of maxillary hypoplasia, DO with a rigid external distraction (RED) system has been used for maxillary advancement; however, DO with internal devices is currently popular. Materials: This article describes DO with an internal device and a RED system in 2 patients with maxillary hypoplasia with oligodontia. The first patient, a young girl, had a concave profile due to maxillary hypoplasia and 9 congenitally missing permanent teeth. At age 11 years 11 months, she received DO with an internal device. The second patient, a boy aged 11 years 7 months, was treated with DO with a RED system. Results: In the girl, the maxilla was advanced 5.0 mm without any dentoalveolar compensation. In the boy, the maxilla was advanced 7.0 mm, but undesirable mesial movement of posterior teeth was observed. Conclusions: DO with internal devices is simpler and more useful than the RED system for maxillary hypoplasia with oligodontia.

Cleft lip and/or palate (CL/P) are caused by many factors. The aim of this study was to investigate the effects of genetic point mutations in CL/P pathogenesis. ICR and AJ strain mice were used. Ethylnitrosourea (ENU) was injected into 10-week-old male mice (G0) intraperitoneally at a dose of 250 mg/kg. The males were bred with two untreated virgin females of the same strain on day 100 after injection. The uterine contents (G1) of one female were examined on day 18.5 of pregnancy. From the other female, the offspring were delivered naturally, and F3 mice (G3) were also examined in the same way. In ICR strain mice, cleft palate only (CPO) was increased in both the G1 and G3. The frequency was significantly higher in the G3 than the G1 generation. Cleft lip was not observed. In AJ strain mice, CL/P increased in both the G1 and G3. In the G3, the frequency of CPO was increased significantly. Genes related to CPO may be recessive in phenotype. CPO and cleft lip differ from a genetic viewpoint. Point mutations play a significant role in cleft lip and palate.

Objective: To identify the location of human β-defensin-2 and human α-defensins in odontogenic keratocysts and radicular cysts. Patients and Methods: Immunohistochemical staining of tissue sections from enucleation specimens of 14 patients with odontogenic keratocysts and 19 patients with radicular cysts, and from normal oral mucosa from 2 healthy volunteers was done using mouse monoclonal antibodies against human cytokeratin 5/6 and rabbit polyclonal antibodies against human β-defensin-2. Results: Human β-defensin-2 immunoreactivity was evident in 10 of 14 odontogenic keratocysts (72%), and in 7 of 19 radicular cysts (37%). Signals for human β-defensin-2 were detected in the cytoplasm of keratinocytes in the parakeratinised and granular layers in both types of cysts. The localisation of human Pdefensin-2 did not differ between cysts and normal mucosa. Human α-defensin immunoreactivity was detected in both types of cysts, but not in keratinocytes. Signals for human α-defensins were detected in the cytoplasm of neutrophils in the lamina propria. Conclusion: Many cysts generate human β-defensin-2 and human α-defensins suggesting that similar defence mechanisms utilising defensins exist in both types of cysts, as well as in normal oral mucosa.

Objective To evaluate the motion of the face and jaw of patients with cleft lip and palate, facial palsy, and in patients after reconstruction, a motion-analyzing system was developed. The aim of this article was to investigate the accuracy of this system and the possibility of clinical application. Methods Markers of 1 to 2 mm were placed on the face, and motion images were obtained by three digital video cameras controlled by a synchronizer and recorded on digital video tape. The image was processed on a personal computer. The markers were automatically tracked across the image sequences, and their three-dimensional coordinates were then calculated. Main Outcome Measures System accuracy was investigated using a positioning actuator with high accuracy and a known object. In three patients with bilateral cleft lip and palate, lip pursing was analyzed using the aforesaid method. Results and Conclusions The mean differences from the known values to the distances between the tracked sample points and to the mobile distances of the sample points per frame were 0.24 to 0.36 mm and 0.02 to 0.05 mm, respectively. Both results were similar regardless of the mobile speed or direction. In five repeated measurements, the mean differences from the known values as for the distances and the mobile speed ranged from 0.19 to 0.38 mm and from 0.00 to 0.07 mm, respectively. Examination of three patients with bilateral cleft lip and palate indicated the possibility that lip movement could be successfully analyzed using the present system.

Immunosuppressant drugs are currently required by transplant recipients for the remainder of their lives, despite the many adverse effects associated with these therapies. Acute osteoporosis is one such effect, and a reproducible osteoporosis model has been established through the administration of the immunosuppressant drug FK506 in rats. The cause of this osteoporosis has been shown to be abnormal osteoclast proliferation, altering the process of bone remodeling. However, the reasons why FK506 induces osteoclast proliferation and whether this process is mediated by cytokine changes or an increase in bone resorption factors have been unclear. An investigation was therefore conducted focusing on the recent discoveries of osteoclast differentiation factor (ODF) and osteoclastogenesis inhibitory factor (OCIF). These factors led to elucidation of the osteoclast differentiation-maturation mechanism. An osteoporosis model was produced in rats utilizing intramuscular FK506 injection (1 mg/kg) for 28 consecutive days. Trabecular bone resorption was observed inferior to enchondral ossification in the FK506 group, and tartrate resistant acid phosphatase (TRAP) staining revealed a clear increase in osteoclasts at the site of enchondral ossification, relative to the control group. Real-time PCR and in situ hybridization (ISH) demonstrated minimal differences in OCIF expression between control and the treatment groups. However, Real-time PCR revealed clearly increased ODF expression in the treatment group. ODF expression was also shown to be increased in the treatment group using ISH. This was histologically consistent with a region of osteoclast proliferation inferior to enchondral ossification. The results of this study support the hypothesis that FK506-mediated osteoporosis occurs by action of the drug on osteoclasts, promoting expression of ODF messenger ribonucleic acid (mRNA) and thus prompting osteoclast differentiation and maturation.

Objective: To establish 3-dimensional measurement norms for the lips of young Japanese adults. Patients and Methods: Landmarks of the lip and face of 80 volunteers were measured on facial plaster models using a contact-type high accuracy 3-dimensional digitiser The lip morphology was analysed by measuring 3-dimensional distances, 3-dimensional angles, and some additional morphological parameters. Several measurement items were compared with Caucasian Americans using Farkas' data. Results: The mean linear distances between the landmarks in men were longer than those in women, except for the upper and lower vermilion height. The upper vermilion was thinner than the lower vermilion in the frontal view, but they were almost the same thickness in linear distance. There were some correlations between the size of the lips and that of the middle and lower face. When compared with Caucasian American standards, the vermilion height and the width of the philtrum were greater, but the cutaneous lip height was shorter in Japanese. The length of the labial fissure was similar in both races. Conclusion: It may be useful for surgery of the maxillofacial region to consider racial differences in normal measurements.

An osteoblastoma of the mandible in a 19-year-old woman is reported. By means of immunohistochemical analysis, the expression of bone morphogenetic proteins, osteopontin, and osteocalcin are described and illustrated.

Purpose: To clarify the involvement of bone morphogenetic proteins (BMPs) in the proliferation and differentiation of osteo/chondrogenic cells during the process of bone formation from grafted periosteum. Material and Methods: Tibial periosteum of young Japanese white rabbits was grafted into suprahyoid muscles and removed after 7, 9, 14 or 21 days. BMP-2, -4, proliferative cell nucleus antigen (PCNA) immunoreaction and Alcian blue staining in grafted periosteum was then sought microscopically. Results: PCNA positive cells in the grafted periosteum expressed BMP-2 at 7 days. These cells differentiated into chondroblasts that expressed BMP-2 and Alcian blue at 9 days. After 14 days, cartilage formation was seen, and BMP-2 and -4 expressions were observed in mature and hypertrophic chondrocytes. Endochondral ossification was observed at 21 days and osteoblasts showed both BMP-2 and -4 expression. Conclusion: Both BMP-2 and -4 appear to play regulatory roles in the process of endochondral ossification from grafted periosteum, due to their involvement in the proliferation and differentiation into chondrogenic and osteogenic cells.

Introduction: The purpose of this study was to assess the three-dimensional (3-D) facial and alveolar morphology of patients with unilateral clefts of lip, alveolus and palate by means of a computer-aided diagnosis system. Patients and Methods: Maxillary orthopaedic treatment was performed using soft/hard acrylic plates (Hotz’s) within 2 weeks of birth. The nasolabial and alveolar morphology of 15 patients was evaluated before orthopaedic treatment (2 weeks of age) and before cheiloplasty (3 months of age). Nasolabial form was measured using a 3-D optical scanner. Twenty-one landmarks were extracted from the data and analysed linearly and angularly. Alveolar forms were measured with a high-accuracy contact-type 3-D digitizer on plaster casts. Seven landmarks were digitized and analysed linearly and angularly. Results: Some growth was observed in the intercanthal distance, alar width, intercommissural width, and height of the lip. There was little change in the width of the cleft lip or displacement of the columella base, while the alveolar cleft narrowed. Conclusion: Presurgical orthopaedics reduces cleft width and makes subsequent surgery easier.

Objective: To determine by immunohistochemical means the localization of human β-defensin-2, a peptide with antimicrobial activity, in oral lichen planus. The release of histamine from mast cells elicited by human β-defensin-2 was also investigated. Materials and Methods: Biopsy specimens of oral lichen planus, synthetic human β-defensin-2, human α-defensin-1, and mast cells isolated from Sprague Dawley rats were used. Tissue sections were embedded in paraffin and immunostained by the streptavidin-biotin-coupled peroxidase method. Isolated rat mast cells were injected with synthetic human β-defensin-2 and human α-defensin-1. Evans blue dye was injected into the tail vein of Sprague Dawley rats, followed by various doses of histamine, human β-defensin-2, human α- defensin-1, and saline. Results: Epithelial cells in lichen planus from the corneal layer to the spinous layer, were positively stained with anti-human β-defensin-2 antibody. Mast cells in subepithelial areas were also stained by anti-human β-defensin-2 antibody. Human β-defensin-2-induced histamine release from the isolated rat mast cells occurs in a dose-dependent manner. When human β-defensin-2 was injected into rat skin intradermally, the vascular permeability increased. These responses were completely abolished upon injection of the antihistamine drug diphenhydramine hydrochloride. Conclusion: Human β-defensin-2 and histamine may play an important role in the formation of oral lichen planus.

Connective tissue growth factor (CTGF) is known to be a multifunctional growth factor that is overexpressed in several types of malignancies. In this study, effects of CTGF gene overexpression on the phenotypes of oral squamous cell carcinoma cells were investigated by using a cell line with undetectable endogenous CTGF expression. Surprisingly, our results indicated that CTGF-overexpressed clones were characterized by attenuated cell growth and less potent tumorigenicity, with coincidental downregulation of prothymosin alpha gene. Although CTGF is known to promote cell proliferation in mesenchymal cells, our present results suggest that CTGF acts as a negative regulator of the cell growth in oral squamous cell carcinoma possibly through its interaction with growth modifiers inside the cell.

Vascular endothelial growth factor (VEGF) plays a very important role in vessel invasion during the process of endochondral ossification. Although grafted periosteum shows endochondral ossification, little is known about the role of VEGF in this process. Additionally, some reports have suggested that bone morphogenetic protein-4 (BMP-4) and VEGF work cooperatively in some way during endochondral ossification. In the present study we investigated, using immunohistochemical, histochemical, ultrastructural, and radiographic techniques, the role of VEGF and BMP-4 in bone formation from periosteal grafts taken from Japanese white rabbit tibia. By 14 days after grafting, fibroblasts in the grafted periosteum differentiated into chondrocytes to form cartilage. Some chondrocytes showed VEGF expression. Subsequent vessel invasion into cartilage from the VEGF immunopositive area coincided with commencement of endochondral ossification. Cartilage was replaced by newly formed bone by 35 days. Soft X-ray findings indicated calcification at 35 days after grafting. Chondrocytes and cartilage matrix were both BMP-4 positive. In newly formed bone, BMP-4 and VEGF expression was demonstrated in osteoblasts. These findings suggest that VEGF are related to vascular invasion into the grafted periosteum during endochondral ossification. Further, VEGF production in chondrocytes and osteoblasts may be related to BMP-4 production.

Connective tissue growth factor (CTGF) is known to be a potent angiogenic factor. Here we investigated how CTGF and matrix metalloproteinases (MMPs) are involved in the early stage of hypoxia-induced angiogenesis using human breast cancer cell line, MDA231, and vascular endothelial cells. Hypoxic stimulation (5% O(2)) of MDA231 cells increased their steady-state level of ctgf mRNA by approximately 2-fold within 1.5 h, and the levels remained at a plateau up to 6 h, and then decreased by 12 h as compared with the cells cultured under the normoxic condition. Membrane-type 1 MMP (MT1-MMP) mRNA levels was also increased within a few hours of the exposure to hypoxia. Indeed, ELISA revealed that the CTGF protein/cell in medium conditioned by MDA231 cells exposed to hypoxia was maximally greater at 24 h than in the medium from normoxic cultures and that the secretion rate (supernatant CTGF/cell layer CTGF) increased in a time-dependent manner from 24 to 72 h of hypoxic exposure. Hypoxic induction of CTGF was also confirmed by immunohistochemical analyses. Furthermore, zymogram analysis revealed that the production of active MMP-9 was also induced in MDA231 cells incubated under hypoxic conditions. Finally, we found that recombinant CTGF also increased the expression of a number of metalloproteinases that play a role in the vascular invasive processes and decreased the expression of tissue inhibitors of metalloproteinases by vascular endothelial cells. These findings suggest that hypoxia stimulates MDA231 cells to release CTGF as an angiogenic modulator, which initiates the invasive angiogenesis cascade by modulating the balance of extracellular matrix synthesis and degradation via MMPs secreted by endothelial cells in response to CTGF. This cascade may play critical roles in the hypoxia-induced neovascularization that accompanies tumor invasion in vivo.

Rheumatoid arthritis (RA) is a disorder of uncertain etiology characterized by immune-mediated chronic inflammatory synovitis, joint effusion, and synovial proliferation showing a prolonged course of repeating remission and exacerbation periods.1 The progression of RA affects many peripheral joints, especially those in the hands and feet, cervical spine, knees, and hips, manifesting as joint destruction, deformity, and/or ankylosis. Synovectomy and/or artificial joint replacement surgery is often performed for the relief of articular pain and functional joint disorders. Of these, peripheral articular manifestations on the cervical spine, cricoarytenoid joint, and temporomandibular joint (TMJ) are the most vexing problems for anesthesiologists. The restricted neck flexion and extension, increased risk of cord compression,2 narrowed glottic aperture,3 increased risk of laryngeal upper airway obstruction,4,5 restricted mouth opening, and acquired retrognathia6 can be potential hazards for general anesthesia, because of the difficulty in airway maintenance by a face mask and in conventional endotracheal intubation using direct laryngoscopy. The extra-articular manifestations of RA, such as pericardial diseases,6,7 restrictive and/or obstructive pulmonary disorders,6,7 neurologic symptoms,8 anemia,6,7 and/or systemic vasculitis6,9 pose additional problems for general anesthesia. The medications prescribed for RA, such as nonsteroidal anti-inflammatory drugs (NSAIDs), long-term corticosteroids, and antirheumatic drugs, may affect platelet function,9 irritate gastrointestinal mucosa,6 confer susceptibility to infectious diseases,9 and/or affect liver and kidney function.6,7 General anesthesia procedures for the management of RA patients have been relatively well documented.6-9 However, no article has referred to the perioperative management of TMJ replacement surgery, including preoperative assessment and considerations and management during and after the surgery, in either RA or non-RA populations. One of the expected sequelae of bilateral total TMJ replacement surgery has been reported to be postoperative facial and jaw swelling.10 In this article, we retrospectively describe the cases of 4 patients with TMJ destruction associated with RA (3 of 4 cases had obstructive sleep apnea [OSA]) who were scheduled for the bilateral total replacement of TMJs. Specific considerations for the anesthetic management of these patients with progressive RA undergoing surgical intervention to the TMJs are discussed. *Assistant Professor, Department of Dental Anesthesiology

To determine whether grafted young periosteum can induce new bone formation in elderly patients, this preliminary study evaluated cell proliferation and differentiation in xenogeneic periosteal grafts in old rats radiographically, histologically, and immunohistochemically. Periosteum harvested from the tibia of young Japanese white rabbits were grafted into old Sprague-Dawley rats with or without administration of 1.0 mg per kilogram per day immunosuppressant FK506. Autogenous old periosteal tissue grafts were also evaluated as a control. Grafted tissue was extirpated after 7, 14, 21, and 45 days. In the xenogeneic group, proliferative cell nuclear antigen-positive cells were observed 7 days after surgery, which differentiated into chondroblasts with bone morphogenetic protein-2 expression and finally formed cartilage by 14 days. Endochondral ossification was observed at 21 days, and bone replacement was completed by 45 days. No osteogenic cell activity was observed in the two other groups. Xenogeneic young periosteum thus maintained its osteogenic/ chondrogenic potentiality in older rats.

Objective: To investigate the release of histamine from rat mast cells elicited by human α-defensin-1 contained in jaw cyst fluid. Patients and Methods: Human α-defensin-1 was purified by reversed-phase high-performance liquid chromatography from human jaw cyst fluid, and the mast cells were collected from the peritoneal cavities of rats. Results: Histamine release was induced in isolated rat mast cells by both extracted and synthetic human α-defensin-1. Changes in vascular permeability induced by human α-defensin-1 were also estimated by the passive skin test in rats. Extracted and synthetic human α-defensin-1 induced histamine release from isolated rat mast cells in a dose-dependent manner over a concentration range of 1 to 15 μg/mL; the histamine release induced by the extracted and synthetic human α-defensin-1 (15 μg/mL) was 39.4 ± 2.3 and 37.6 ± 1.8%, respectively. When human α-defensin-1 was injected into rat skin intradermally, the vascular permeability increased. This response occurred with as little as 0.1 μg of human α-defensin-1, and a strong response was observed with 0.5 and 1.0 μg. These responses were completely abolished upon injection with 1 μg of the antihistamine drug diphenhydramine hydrochloride. Conclusion: Human α-defensin-1 in human jaw cyst fluid can act as an inducer of histamine release from mast cells both in vivo and in vitro.

OBJECTIVES AND DESIGN: It has been previously reported that α-defensin (HNPs) and β-defensin-2 (HBD-2) peptides with antifungal and cytotoxic activities can be detected in oral carcinomas and the saliva of patients with oral carcinomas. The present study investigated the presence of HNPs and HBD-2 in oral epithelia with candidiasis. MATERIALS AND METHODS: Tissue sections (4 μm) were prepared from biopsy and surgically removed specimens diagnosed as oral candidiasis (n=10). The sections were examined immunohistochemically with antibodies directed against HNPs and HBD-2. RESULTS: Tissue sections of oral candidiasis were immunostained with antidefensin antibodies. Neutrophils in the inflamed lamina propria were positively immunostained with anti-HNPs antibody. The cytoplasm of cells in the upper spinous layer, in the lower spinous layer and in the parakeratinized layer of buccal epithelia with candidiasis was immunostained intensely with anti-HBD-2 antibody. In contrast, the expression of HBD-2 in the normal spinous layer was much weaker than that in oral candidiasis. No signals of HNPs were found in normal buccal epithelium. CONCLUSION: Buccal specimens from individuals with oral candidiasis show greater levels of expression of both HNPs and HBD-2. There might be a dual protection manner by defensins against fungal inflammation in infected buccal epithelia locally. Generally, HBD-2 signals have been found everywhere in the buccal epithelium; however, in an infected area, the signal intensity of HBD-2 has increased. HNPs signals have not been found in the normal buccal epithelium; however, HNPs signals have increased when the infection occurred.