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Granular-shaped carbonate apatite (CO3Ap) bone substitute has demonstrated good clinical results, especially in the bone augmentation procedure, because it has high osteoconductivity and can be replaced by a new bone. This study aimed to evaluate the potential usefulness of CO3Ap bone substitute for alveolar ridge preservation of beagle dogs during the 9-week follow-up period after surgery. Reconstruction of extraction sockets with CO3Ap granules (CO3Ap group) allowed approximately 1.0 mm higher alveolar bone height than that with a blood clot (control group) at 4 and 9 weeks after surgery. Additionally, the measured area at the buccal aspect of the alveolar bone in the CO3Ap group was approximately 2 mm2 larger than that in the control group at 4 and 9 weeks after surgery. Continuity from the buccal side to palatal side of the alveolar bone was restored, maintaining its contour at 4 weeks after surgery. The amount of bony tissue, especially the amount of mature bone, in the alveolar part was statistically larger than that of control group at 9 weeks after surgery even though complete replacement of CO3Ap granules with new bone was not observed within 9 weeks. In conclusion, CO3Ap granules have good potential for alveolar ridge preservation.

Calcium hydroxide (Ca (OH)2) paste is widely used as a provisional root canal filling material. This report describes a case of inferior alveolar nerve paresthesia caused by overfilling of Ca (OH)2 paste. A 53-year-old woman had previously undergone root canal filling in the left lower second molar with Ca (OH)2 paste. Immediately after the treatment, she felt sharp pain and paralysis in the left lower lip and chin area and visited our hospital four days later. X-ray examination showed a large radiopacity associated with the left mandibular canal and around the bone marrow from the lower left second premolar to the lower left third molar. We planned a tooth extraction and surgical removal of the Ca (OH)2 paste. However, she declined surgery, and we performed conservative treatment with administration of vitamin B12 and root canal therapy. Approximately one year after her first visit, we filled the root canal with gutta-percha and sealer. The tooth was preserved and paresthesia of the left lower lip and chin largely decreased except for a small remaining numb area. Therefore, it is important to pay scrupulous attention to ensure no extrusion of Ca (OH)2 paste beyond the root apex to avoid complications.

Dental implants made of titanium are commonly used. Although titanium implants succeed by osseointegration with bone, the detailed molecular mechanism of osseointegration is unclear. To clarify the involvement of microRNA (miRNA) in the acquisition of osseointegration on titanium, here we compared the miRNA expression profiles of mouse osteoblast-like cells (MC3T3-E1) cultured on titanium-, gold-, and stainless steel-coating glass dishes by microarray analysis. Three kinds of metals, namely titanium, gold, and stainless steel, were coated on the surface of the glass dishes by sputtering with similar roughness and shape of their surface. After MC3T3-E1 cells were cultured on the dishes without coating or coating with titanium, gold, or stainless steel for 6 h, total RNA was extracted, and miRNA expression was analyzed by microarray. To confirm the expression of the selected miRNA during osteogenic differentiation of MC3T3-E1 cells, real-time PCR analysis was performed. Furthermore, the effects of selected miRNA were examined by ectopic overexpression in MC3T3-E1 cells. The microarray analysis revealed that the expressions of miR-155-5p and miR-7023-3p were significantly increased in MC3T3-E1 cells cultured on titanium-coating glass dishes, compared to non-coating, gold-, and stainless steel-coating glass dishes. Interestingly, miR-155-5p was upregulated during osteogenic differentiation of MC3T3-E1 cells. Furthermore, overexpression of miR-155-5p enhanced the expression of Runx2 and Col1a1. In this study, miR-155-5p may be involved in the acquisition of osseointegration on titanium implant via upregulating osteogenic differentiation-related genes.

The present study investigated the osteoclast differentiation potential and paracrine effects of osteoclasts on osteoblast differentiation when the cells were cultured directly on two bone substitutes (BSs): deproteinized bovine bone mineral (DBBM) and carbonate apatite (CO3 Ap). Human primary osteoclasts cultured on the BSs were assessed by tartrate-resistant acid phosphatase (TRAP) and actin ring staining. Thereafter, the mRNA levels of osteoclastic differentiation markers were quantified by real-time PCR. Osteoblast behaviors in response to conditioned media collected from osteoclast cultures were investigated. Interestingly, mature osteoclasts were occasionally observed on the surface of the CO3 Ap granules, whereas very few and small osteoclasts were observed on DBBM. Similarly, real-time PCR analysis showed higher mRNA levels of osteoclast markers, including cathepsin K and TRAP, in the cells cultured on CO3 Ap than in those cultured on DBBM. Furthermore, compared to DBBM, CO3 Ap promoted osteoblast differentiation in human primary osteoblasts, whereas few paracrine effects of osteoclasts cultured with either BS were observed on the osteoblast differentiation potential. These limited results showed that CO3 Ap provided a favorable surface for osteoclast differentiation, as well as osteoblasts, compared to DBBM in vitro.

This study aimed to create a predictive model for cervical lymph node metastasis (CLNM) in patients with tongue squamous cell carcinoma (SCC) based on radiomics features detected by [A total of 40 patients with tongue SCC who underwentSix radiomics features were selected from LASSO analysis. The average values of the area under the curve (AUC), accuracy, sensitivity, and specificity of radiomics analysis for predicting CLNM fromThe

Background: Oral allergy syndrome (OAS) is a type of food allergy that manifests as hypersensitivity symptoms of the oropharyngeal mucosa on ingestion of specific foods with oral findings resembling herpetic gingivostomatitis. Symptoms can also appear in parts other than the oral cavity. There are a few reports of OAS caused by consuming radishes in the literature.Case presentation: A 31-year-old male presented to our department with stomatitis and pharyngeal pain. The patient did not have a history of allergy, and herpetic gingivostomatitis was suspected. He was admitted to the emergency room of our hospital after complaining of oral and epigastric pain a few days later. On admission, symptoms noted were similar to those when he first visited our department. After detailed history taking, he reported consuming raw Japanese radish frequently, which gave rise to his symptoms. Radish was strongly suspected as the causative allergen. The skin prick test result was positive, confirming the diagnosis of OAS.Conclusions: OAS can be diagnosed easily once the food causing symptoms is identified. However, guiding the diagnosis can be challenging if the patient is unaware of food allergies. Upon encountering widespread erosion in the oral cavity, it is essential to consider OAS as the possible cause.

Programmed cell death-1 (PD-1) and its ligand programmed cell death 1 ligand 1 (PD-L1) are immune checkpoint inhibitors that play an important role in the host immune avoidance mechanism of tumors. The relationship between PD-L1 expression and malignancy has been reported in various types of cancer, such as lung and gastric cancer. In addition, epithelial-mesenchymal transition (EMT) of cancer cells is deeply involved in the invasion and metastasis of cancer. It has been reported that zinc finger E-box binding homeobox 1 (ZEB-1), an EMT inducer, contributes to metastasis in pancreatic and colon cancer. The present study aimed to investigate the relationship between the expression patterns of two markers, PD-L1 and ZEB-1, and clinicopathological characteristics and prognosis of oral squamous cell carcinoma (OSCC). Biopsy or surgical excision specimens from 169 patients with OSCC were used in the present study. Immunohistochemical staining with monoclonal anti-PD-L1 antibody and anti-ZEB-1 antibody was conducted. Cases with >1% tumor cells positive for PD-L1 and those with >10% tumor cells positive for ZEB-1 were considered positive, respectively. The findings revealed that individual expression of PD-L1 and ZEB-1 in OSCC was not associated with tumor size, degree of differentiation or Yamamoto-Kohama invasion pattern classification. However, co-expression of PD-L1 and ZEB-1 was associated with higher cervical lymph node metastasis and a lower survival rate. In conclusion, the results of the present study indicated that co-expression of PD-L1 and ZEB-1 could serve as a potential marker for the prognosis of patients with OSCC.

Beta-tricalcium phosphate granular cement (β-TCP GC), consisting of β-TCP granules and an acidic calcium phosphate (Ca-P) solution, shows promise in the reconstruction of bone defects as it sets to form interconnected porous structures, that is, β-TCP granules are bridged with dicalcium phosphate dihydrate (DCPD) crystals. In this study, the effects of acidic Ca-P solution concentration (0-600 mmol/L) on the setting reaction and tissue response to β-TCP GC were investigated. The β-TCP GC set upon mixing with its liquid phase, based on the formation of DCPD crystals, which bridged β-TCP granules to one another. Diametral tensile strength of the set β-TCP GC was relatively the same, at ∼0.6 MPa, when the Ca-P concentration was 20-600 mmol/L. Due to the setting ability, reconstruction of the rat's calvarial bone defect using β-TCP GC with 20, 200, and 600 mmol/L Ca-P solution was much easier compared to that with β-TCP granules without setting ability. Four weeks after the reconstruction, the amount of new bone was the same, ∼17% in both β-TCP GC and β-TCP granules groups. Cellular response to β-TCP granules and β-TCP GC using the 20 mmol/L acidic Ca-P solution was almost the same. However, β-TCP GC using the 200 and 600 mmol/L acidic Ca-P solution showed a more severe inflammatory reaction. It is concluded, therefore, that β-TCP GC, using the 20 mmol/L acidic Ca-P solution, is recommended as this concentration allows surgical techniques to be performed easily and provides good mechanical strength, and the similar cellular response to β-TCP granules. © 2019 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater 108B:22-29, 2020.

Lower third molar extraction is the most common surgical treatment among routine dental and oral surgical procedures. while the surgical procedures for lower third molar extraction are well established, the difficulty of tooth extraction and the frequency of postoperative complications differ depending on the patient’s background. To establish a management protocol for the lower third molars, the prevalence of and risk factors for postoperative complications after lower third molar extraction were investigated in a large number of Japanese patients in a multicenter prospective study. During 6 consecutive months in 2020, 1826 lower third molar extractions were performed at the 20 participating institutions. The medical records of the patients were reviewed, and relevant data were extracted. The prevalence of and risk factors for postoperative complications were analyzed. The prevalence of postoperative complications after lower third molar extraction was 10.0%. Multivariate analysis indicated that age (≤32 vs >32, odds ratio [OR]: 1.428, 95% confidence interval [95% CI]: 1.040–1.962, P < .05), the radiographic anatomical relationship between the tooth roots and mandibular canal (overlapping of the roots and canal vs no close anatomical relationship between the roots and the superior border of the canal, OR: 2.078, 95% CI: 1.333–3.238, P < .01; overlapping of the roots and canal vs roots impinging on the superior border of the canal, OR: 1.599, 95% CI: 1.050–2.435, P < .05), and impaction depth according to the Pell and Gregory classification (position C vs position A, OR: 3.7622, 95% CI: 2.079–6.310, P < .001; position C vs position B, OR: 2.574, 95% CI: 1.574–4.210, P < .001) are significant independent risk factors for postoperative complications after lower third molar extraction. These results suggested that higher age and a deeply impacted tooth might be significant independent risk factors for postoperative complications after lower third molar extraction.

Carbonate apatite (CO3 Ap) block can be used as a bone substitute because it can be remodeled to new natural bone in a manner conforming with the bone remodeling process. Among the many porous structures available, honeycomb (HC) structure is advantageous for rapid replacement of CO3 Ap to bone. In this study, the feasibility to fabricate a CO3 Ap HC was studied, along with its initial tissue response in rabbit femur bone defect. First, a mixture of Ca(OH)2 and a wax-based binder was extruded from a HC mold. Then the fabricated HC was heated for binder removal and carbonation at 450°C in a mixed O2 -CO2 atmosphere, forming a CaCO3 HC. When the CaCO3 HC was immersed in 1 mol/L Na3 PO4 solution at 80°C for 7 days, its composition changed from CaCO3 to CO3 Ap, maintaining the structure of the original CaCO3 HC. Compressive strengths of the CaCO3 and CO3 Ap HCs were 65.2 ± 7.4 MPa and 88.7 ± 4.7 MPa, respectively. When the rabbit femur bone defect was reconstructed with the CO3 Ap HC, new bone penetrated the CO3 Ap HC completely. Osteoclasts and osteoblasts were found on the surface of the newly formed bone and osteocytes were also found in the newly formed bone, indicating ongoing bone remodeling. Furthermore, blood vessels were formed inside the pores of CO3 Ap HC. Therefore, CO3 Ap HC has good potential as an ideal bone substitute. © 2019 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 107A: 1014-1020, 2019.

Cancer stem cells (CSCs) exhibit self-replication, self-differentiation, drug resistance and immune evasion activities. In recent years CSCs have become increasingly important for the treatment of malignant tumors. CSCs express specific markers, including cluster of differentiation (CD)44, CD44 variant 9 (CD44v9), ATP-binding cassette sub-family G member 2 (ABCG2), CD24, B lymphoma Mo-MLV insertion region 1 homolog (BMI-1) and aldehyde dehydrogenase 1 (ALDH1). However, the prognostic value of their expression in patients with oral squamous cell carcinoma (OSCC) are not well known. The present study evaluated these markers in stage I and II patients with OSCC and examined the association between T classification, histological differentiation, classification of invasion mode, lymph node metastasis and disease-free survival rate. Tissue specimens were obtained from 70 patients with stage I or II OSCC following either surgery or biopsy. Immunohistochemistry was performed and positive staining was defiend as 10% positive cells. CD44 and CD44v9 expressions were strongly detected in all OSCC tissues compared with normal epithelial cells. A total of 22 (31.4%) cases expressed ABCG2 and there was a significant association between ABCG2 expression and invasion. A total of 41 cases (59.0%) expressed CD24 and there was a significant association between CD24 expression and invasion. A total of 33 cases (47.1%) expressed BMI-1 and there was a significant association between BMI-1 expression and the disease-free survival rate. A total of 18 cases (25.7%) expressed ALDH1. Although there was no association between ALDH1 expression and T classification, there were significant associations between ALDH1 expression and histological differentiation, invasion mode, metastasis and the disease-free survival rate. Multivariate analysis revealed that ALDH1 expression was the only prognostic factor for disease-free survival rate. The results of the present study suggest that the positivity of ALDH1 detected in patients with OSCC correlates with the number of cells undergoing epithelial mesenchymal transition and metastasis. These findings indicated that the expression of ALDH1 may be an effective prognostic marker indicating the survival of patients with stage I and II OSCC.

The aim of this study was to evaluate clinical outcomes of staged sinus floor elevation (SFE) using novel low-crystalline carbonate apatite (CO3Ap) granules. Patients who needed SFE for implant placement were recruited into this clinical trial. A staged procedure (lateral window technique using CO3Ap granules, followed by implant placement after 7 ± 2 months) was employed in 13 patients. Bone-height increase and insertion torque values (ITVs) were assessed along with histological evaluation. The survival and success rates of 3-year functioning implants were also evaluated. Mean of bone-height increase after SFE using CO3Ap granules was 7.2 ± 2.5 mm and this increase allowed implant placement in all cases (17 implants). Mean of ITV was 25.1 ± 13.2 Ncm and primary stability was achieved successfully in all cases. Histological analyses revealed mature new bone formation (36.8 ± 17.3%) and residual CO3Ap granules (16.2 ± 10.1%) in the compartment after SFE. The survival and success rates after 3-year functional loading were 100% and no complications were found. These results clearly indicate the clinical usefulness of CO3Ap granules for SFE.

Objective The aim of this study is to reveal a clinical evaluation of single non-contact Nd: YAG (the neodymium: yttrium-aluminium-garnet laser) photocoagulation irradiation treatment of venous malformation (VM) in the oral region. Patients and methods Seventeen patients with 21 VMs in the oral region who visited our hospital from 2010 to 2015 were subjected. The patients consisted of six males and 11 females, ranging in age from 27 to 71 years old. The VM locations included tongue, buccal mucosa, and lip sites. Their sizes ranged from 4 to 24 mm along the major axis direction, and the thickness ranged from 4 to 13 mm. All patients were treated with single non-contact irradiation using the Nd: YAG laser under local anaesthesia. Results All cases, including the patients with deep-seated lesions, healed after one-time irradiation with the laser beam. Serious complications have not occurred in patients receiving the treatment. There was no recurrence of VMs for at least one year after treatment. Conclusions Using single non-contact irradiation treatment, we obtained good results without clinical complications. The application of the Nd: YAG laser in treatment of lesions has the advantages of no bleeding, minimal scarring, and a short operative time. These results suggest that Nd: YAG laser treatment by a single non-contact irradiation is effective against VMs of thicknesses 13 mm or less in the oral region.

A case of tophaceous pseudogout (i.e., calcium pyrophosphate dihydrate (CPPD) crystal deposition disease) in the temporomandibular joint (TMJ), extending to the base of the skull, is reported. A 38-year-old man was referred to the hospital with mild pain in the right chin and tip of the tongue. Panoramic radiography showed a large calcified mass around the right TMJ. Computed tomography imaging revealed a large, granular, calcified mass surrounding the right condylar head and extending to the base of the skull. The mass was clinically and radiographically suspected to be a pseudogout lesion. A biopsy specimen was collected under general anaesthesia to confirm the diagnosis. On histology, the mass was found to contain deposits of numerous rod-shaped and rhomboid crystals, which suggested tophaceous pseudogout. The deposits were identified as CPPD crystal deposition, based on analysis by X-ray diffraction and Fourier transform infrared spectroscopy. These two crystallography methods were useful in confirming the diagnosis of CPPD crystal deposition disease in the TMJ.

Carbonate apatite (CO3 Ap) granules are known to show good osteoconductivity and replaced to new bone. On the other hand, it is well known that a porous structure allows bone tissue to penetrate its pores, and the optimal pore size for bone ingrowth is dependent on the composition and structure of the scaffold material. Therefore, the aim of this study was to fabricate various porous CO3 Ap granules through a two-step dissolution-precipitation reaction using CaSO4 as a precursor and 30-, 50-, 120-, and 205-μm diameter microfibers as porogen and to find the optimal pore size of CO3 Ap. Porous CO3 Ap granules were successfully fabricated with pore size 8.2-18.7% smaller than the size of the original fiber porogen. Two weeks after the reconstruction of rabbit calvarial bone defects using porous CO3 Ap granules, the largest amount of mature bone was seen to be formed inside the pores of CO3 Ap (120) [porous CO3 Ap granules made using 120-μm microfiber] followed by CO3 Ap (50) and CO3 Ap (30). At 4 and 8 weeks, no statistically significant difference was observed based on the pore size, even though largest amount of mature bone was formed in case of CO3 Ap (120). It is concluded, therefore, that the optimal pore size of the CO3 Ap is that of CO3 Ap (120), which is 85 μm.

Purpose The purpose of this study was to elucidate the efficacy and safety of carbonate apatite (CO3Ap) granules in 2-stage sinus floor augmentation through the radiographic and histomorphometric assessment of bone biopsy specimens. Methods Two-stage sinus floor augmentation was performed on 13 patients with a total of 17 implants. Radiographic assessment using panoramic radiographs was performed immediately after augmentation and was also performed 2 additional times, at 7±2 months and 18±2 months post-augmentation, respectively. Bone biopsy specimens taken from planned implant placement sites underwent micro-computed tomography, after which histological sections were prepared. Results Postoperative healing of the sinus floor augmentation was uneventful in all cases. The mean preoperative residual bone height was 3.5±1.3 mm, and this was increased to 13.3±1.7 mm by augmentation with the CO3Ap granules. The mean height of the augmented site had decreased to 10.7±1.9 mm by 7±2 months after augmentation; however, implants with lengths in the range of 6.5 to 11.5 mm could still be placed. The mean height of the augmented site had decreased to 9.6±1.4 mm by 18±2 months post-augmentation. No implant failure or complications were observed. Few inflammatory cells or foreign body giant cells were observed in the bone biopsy specimens. Although there were individual differences in the amount of new bone detected, new bone was observed to be in direct contact with the CO3Ap granules in all cases, without an intermediate layer of fibrous tissue. The amounts of bone and residual CO3Ap were 33.8%±15.1% and 15.3%±11.9%, respectively. Conclusions In this first demonstration, low-crystalline CO3Ap granules showed excellent biocompatibility, and bone biopsy showed them to be replaced with bone in humans. CO3Ap granules are a useful and safe bone substitute for two-stage sinus floor augmentation. Trial Registration ICTRP Identifier: JPRN-UMIN000019281, Graphical Abstract

Since bone apatite is a carbonate apatite containing carbonate in an apatitic structure, carbonate content may be one of the factors governing the osteoconductivity of apatitic bone substitutes. The aim of this study was to evaluate the effects of carbonate content on the osteoconductivity of apatitic bone substitutes using three commercially available bone substitutes for the reconstruction of alveolar bone defects of a beagle mandible with simultaneous dental implant installation. NEOBONE®, Bio-Oss®, and Cytrans® that contain 0.1 mass%, 5.5 mass%, and 12.0 mass% of carbonate, respectively, were used in this study. The amount of newly formed bone in the upper portion of the alveolar bone defect of the beagle’s mandible was 0.7%, 6.6%, and 39.4% at 4 weeks after surgery and 4.7%, 39.5% and 75.2% at 12 weeks after surgery for NEOBONE®, Bio-Oss®, and Cytrans®, respectively. The results indicate that bone-to-implant contact ratio was the largest for Cytrans®. Additionally, the continuity of the alveolar ridge was restored in the case of Cytrans®, whereas the continuity of the alveolar ridge was not sufficient when using NEOBONE® and Bio-Oss®. Both Cytrans® and Bio-Oss® that has a relatively larger carbonate content in their apatitic structure was resorbed with time. We concluded that carbonate content is one of important factors governing the osteoconductivity of apatitic bone substitutes.

OK-432 is a potent immunotherapy agent for several types of cancer, including oral cancer. We previously reported that OK-432 treatment can induce the production of high levels of IFN-γ from peripheral blood mononuclear cells (PBMCs). Moreover, the IFN-γ production from PBMCs by OK-432 is impaired by conditioned media (CM) from oral cancer cells. To determine the inhibitory mechanism of IFN-γ production by CM, the genes involved in IFN-γ production was retrieved by cDNA microarray analysis. We found that CD40 played a key role in IFN-γ production via IL-12 production. Although the expression levels of CD40 were upregulated by OK-432 treatment in PBMCs, CM inhibited OK-432-induced CD40 expression. These findings suggest that uncertain soluble factor(s) in CM may suppress IFN-γ production via the CD40/CD40L–IL-12 axis in PBMCs., (1) Background: OK-432 is a penicillin-killed, lyophilized formulation of a low-toxicity strain (Su) of Streptococcus pyogenes (Group A). It is a potent immunotherapy agent for several types of cancer, including oral cancer. We previously showed that (i) OK-432 treatment induces a high amount of IFN-γ production from peripheral blood mononuclear cells (PBMCs), and (ii) conditioned medium (CM) from oral cancer cells suppresses both the IFN-γ production and cytotoxic activity of PBMCs driven by OK-432. The aim of this study was to determine the inhibitory mechanism of OK-432-induced IFN-γ production from PBMCs by CM. (2) Methods: We performed cDNA microarray analysis, quantitative RT-PCR, and ELISA to reveal the inhibitory mechanism of CM. (3) Results: We found that CD40 plays a key role in IFN-γ production via IL-12 production. Although OK-432 treatment upregulated the expression levels of the IL-12p40, p35, and CD40 genes, CM from oral cancer cells downregulate these genes. The amount of IFN-γ production by OK-432 treatment was decreased by an anti-CD40 neutralizing antibody. (4) Conclusions: Our study suggests that uncertain soluble factor(s) produced from oral cancer cells may inhibit IFN-γ production from PBMCs via suppressing the CD40/CD40L–IL-12 axis.

Purpose Carbonate apatite (CO3Ap), an inorganic component of human bone, can be fabricated in chemically pure form from calcium carbonate block via a dissolution-precipitation reaction. A first-in-human clinical trial was conducted in which low-crystalline CO3Ap granules were evaluated for safety and efficacy in sinus floor augmentation and simultaneous implant installation. Materials and Methods Procedures were performed in 8 patients (9 implants) with 2 granule sizes: small (300 to 600 μm) and medium (600 to 1,000 μm). Panoramic radiographic assessment was performed immediately after augmentation, 7 ± 2 months after augmentation, 6 ± 2 months after prosthetic loading, and 12 ± 2 months after prosthetic loading. Results Postoperative healing was uniformly uneventful, with no abnormal bleeding, pain, or swelling, and all implants achieved successful osseointegration. The mean residual maxillary molar bone height was 5.2 ± 0.8 mm preoperatively and increased to 14.0 ± 1.9 mm after augmentation. Implants 9.0 to 11.5 mm in length were placed. The post-augmentation height decreased to 12.4 ± 1.3 mm at 7 ± 2 months; after prosthetic loading, it decreased to 11.9 ± 0.8 mm at 6 ± 2 months and 11.7 ± 0.6 mm at 12 ± 2 months. No abnormal bone resorption of the augmented areas was observed, and bone height supporting the implants was maintained. The overall implant survival rate was 100%, with no implant failures or complications during the first year. Conclusions Low-crystalline CO3Ap granules were useful and safe for sinus floor augmentation and simultaneous implant installation, providing a promising bone substitute for dental implant surgery.

We have previously demonstrated that the stromal cell‑derived factor (SDF‑1)/CXCR4 system is involved in the metastasis of head and neck cancer. Additionally, it has been revealed that the blockade of CXCR4 by subcutaneous daily injection with AMD3100, a CXCR4 antagonist, may be effective in preventing metastasis in CXCR4‑related head and neck cancer. Recent investigations have suggested that AMD070, a novel orally bioavailable inhibitor of CXCR4, may be minimally invasive compared with AMD3100. In the present study, we examined the effect of AMD070 on metastasis induced by the SDF‑1/CXCR4 axis in B88‑SDF‑1 oral cancer cells, which express high levels of SDF‑1 and CXCR4. Although treatment with AMD070 did not affect the anchorage‑dependent growth of B88‑SDF‑1 cells, it significantly suppressed the anchorage‑independent growth. Moreover, the SDF‑1/CXCR4‑dependent migration and invasion of B88‑SDF‑1 cells was significantly inhibited following treatment with AMD070. Subsequently, we performed an experimental therapy using AMD070 to prevent the distant metastasis of B88‑SDF‑1 cells in vivo. Daily oral administration of AMD070 significantly inhibited the lung metastasis of B88‑SDF‑1 cells in nude mice. These results indicated that AMD070 could be useful as a novel orally bioavailable inhibitor of oral cancer metastasis.

Objectives: Palatine tonsilloliths incidentally detected on diagnostic imaging should be differentiated from pathologic calcifications to enable correct diagnosis and treatment. The aim of this study is to clarify the prevalence and imaging characteristics of palatine tonsilloliths on panoramic radiographs. Materials and Methods: We retrospectively reviewed 2,244 individuals who underwent pairs of consecutive panoramic radiography and computed tomography (CT) of the head and neck region. The imaging characteristics of palatine tonsilloliths on panoramic radiography were compared with the findings from CT, which was considered the gold standard. Results: Tonsilloliths were detected in 300 (13.4%) and 914 (40.7%) of the 2,244 individuals on panoramic radiographs and CT, respectively. On panoramic radiographs, tonsilloliths were superimposed over the ramus of the mandible at the level coincident with and inferior to the soft palate in 176 (7.8%) and 90 (4.0%) individuals, respectively. Tonsilloliths were also superimposed over the surrounding soft tissue inferior to the body of the mandible, postero-inferior to the angle of the mandible, and posterior to the ramus of the mandible in 33 (1.5%), 26 (1.2%), and 28 (1.3%) individuals, respectively. A significant correlation was observed between the detectability on panoramic radiographs and the size (Spearman r=1.000) and number (Spearman r=0.991) of tonsilloliths, as revealed by CT images. Conclusions: The present results suggest that tonsilloliths are commonly detected on panoramic radiographs. Furthermore, they can be superimposed on both the mandible and the surrounding soft tissue. Clinical Relevance: Clinicians should include tonsilloliths among the differential diagnoses when calcified bodies are detected on panoramic radiographs.

The zygomaticus implant (Brånemark system, Nobel Biocare, Gotebörg, Sweden) was developed for patients with severe bone resorption of the posterior maxilla, which may eliminate or minimize the need for bone grafting. Although the zygomaticus implant has had a remarkable success rate in a difficult patient population, the method requires an advanced surgical technique and carries increased risk of complications, such as the perforation of the orbital floor or infratemporal fossa. Although it is important to have a detailed understanding of the anatomy of the zygomatic bone when performing the installation, there have been few anatomic studies on the zygomatic bone for installation of zygomaticus implants. In this study, we measured the height and thickness of the zygomatic bone for the installation. The thickness at a 90-degree angle point, where the upper margin of the zygomatic arch and the temporal margin of the frontal process of the zygomatic bone intersect and where the apex of the implant penetrates, according to the original method, was 1.8 ± 0.4 mm, which gradually increased inferiorly and anteriorly. In conclusion, the penetration point of the apex of the zygomaticus implant should be located more inferoanterior to the 90-degree angle point, as the thickness of the 90-degree angle point is thinner than the diameter of the implant. Based on these results, we have proposed a newer and safer installation method for the zygomaticus implant using a drill guide, which can be easily made.

Neuroendocrine tumors (NETs) arise from neuroendocrine cells and are mostly observed in the gastrointestinal tract, pancreas, and lungs. NETs in the oral and maxillofacial region are extremely rare. We report a case of a 59-year-old woman with an NET in the mandible. The patient did not show any symptoms except for remarkable swelling and bleeding. The lesion appeared as a radiolucent honeycomb abnormality with bone destruction on panoramic radiography. The histopathologic diagnosis following a biopsy was NET. Contrast-enhanced computed tomography (CT), 18F-fluorodeoxyglucose positron emission computed tomography (18F-FDG PET/CT), and adrenal scintigraphy-labeled meta-iodobenylguanidine were the modalities added to identify the primary site. Multiple lesions were confirmed in the gastrointestinal tract. Endoscopy was performed to identify the lesions, and several lesions were observed protruding from the mucous membranes. However, the endoscopy specimens did not yield an accurate diagnosis because adequate samples were not acquired. Blood and urine tests revealed no functional activity caused by the tumors. Although the origin was not histopathologically confirmed with endoscopy, this patient was situationally diagnosed with nonfunctional NET originating from the duodenum, as demonstrated by the metastases in the mandible.

Carbonate apatite (CO3Ap) is an inorganic component of bone. This study aimed to compare the composition and tissue response to of CO3Ap (CO3Ap-DP) fabricated by the dissolution-precipitation reaction using calcite as a precursor and Bio-Oss®, which is widely used in orthopedic and dental fields as a synthetic bone substitute. X-ray diffraction and Fourier transform infrared results showed that CO3Ap-DP and Bio-Oss® were both B-type carbonate apatite with low crystallinity. The average sizes of CO3Ap-DP and Bio-Oss® granules were 450 ± 58 and 667 ± 168μ m, respectively, and their carbonate contents were 12.1 ± 0.6 and 5.6 ± 0.1 wt%, respectively. CO3Ap-DP had a larger amount of CO3 than Bio-Oss® but higher crystallinity than Bio-Oss®. When a bone defect made at the femur of rabbits was reconstructed with CO3Ap-DP and Bio-Oss®, CO3Ap-DP granules were partially replaced with bone, whereas Bio-Oss® remained at 8 weeks after implantation. CO3Ap-DP granules elicited a significantly larger amount of new bone formation at the cortical bone portion than Bio-Oss® at 4 weeks after the implantation. The results obtained in the present study demonstrated that CO3Ap-DP and Bio-Oss® showed different behavior even though they were both classified as CO3Ap. The CO3 content in CO3Ap played a more important role than the crystallinity of CO3Ap for replacement to bone and high osteoconductivity.

Background : Mandibular reconstruction using a fibular graft is a difficult procedure that requires technical expertise to enable adequate occlusal function and restore an aesthetic appearance. Here we used three-dimensional (3D) sodium chloride (NaCl) models for simulated mandibular reconstruction surgery. This study aimed to reveal the accuracy of mandibular reconstruction with fibular grafts using this model. Methods : Mandibular reconstructions using 3D NaCl models were performed in 5 cases. The maxilla, mandible, and fibular models were developed using computed tomography (CT) data. We performed preoperative cutting and simulation surgery using this model. Angles between the body of the corpus and symphysis were measured from the axial view (n = 4). Angles between the ramus and the body of the corpus were measured from the lateral and axial views (n = 6). These angles were measured on simulated models and postoperative CT images were compared. Results : Differences between the angles measured on the simulated models and postoperative CT images were 0-5°(mean, 1.9°). Conclusions : We were able to reproduce the neomandibles with precise osteotomies using the 3D NaCl models. We believe that simulated mandibular reconstruction surgery using this model might help reduce the number of intraoperative neomandibular segment adjustments.

Salivary gland cancer (SGC) has a comparatively poor prognosis and is prone to frequent recurrence and metastases. Therefore, the development of more effective chemotherapy against SGC is desirable. The aim of the present study was to investigate the antitumour effects of valproic acid (VPA) against SGC in vitro and in vivo. Two human SGC cell lines (HSY and HSG cells) were used in the present study. The effects of VPA on the proliferation of SGC cells in vitro were assessed by MTT assay. Cancer cells treated with VPA were subjected to cell cycle analysis by flow cytometry. In addition, the expression levels of p21 and p27 were examined by real-time RT-PCR to identify the mechanisms of the antitumour effect of VPA on SGC. The effects of VPA on cancer growth in vivo were evaluated in a xenograft model. VPA inhibited the proliferation of SGC cells in a dose-dependent manner in vitro. Degenerated cancer cells were observed at high concentrations of VPA. In the cell cycle analysis, VPA induced cell-growth inhibition and G1 arrest of cell cycle progression in both cancer cell lines in a time- and dose-dependent manner. VPA markedly upregulated the mRNA expression levels of both p21 and p27 in both SGC cell lines in a time-dependent manner. In the xenograft model experiment, VPA treatment markedly inhibited the growth of salivary gland tumours when compared with the growth of the untreated controls. VPA may be a valuable drug in the development of better therapeutic regimens for SGC.

Objective: Clear cell odontogenic carcinoma (CCOC) is a rare malignant odontogenic tumor (MOT) characterized by sheets and lobules of vacuolated and clear cells. To understand the biology of CCOC, we established a new cell line, CCOC-T, with EWSR1-ATF1 fusion gene from a mandible tumor with distant metastasis and characterized this cell line. Materials and methods: To detect the EWSR1-ATF1 fusion gene, we used three CCOC cases, including the present case, by RT-PCR and FISH analysis. We characterized established CCOC-T cells by checking cell growth, invasion and the expression of odontogenic factors and bone-related factors. Moreover, the gene expression profile of CCOC-T cells was examined by microarray analysis. Results: Histologically, the primary tumor was comprised of cords and nests containing clear and squamoid cells separated by fibrous septa. In addition, ameloblastomatous islands with palisaded peripheral cells were observed, indicating probable odontogenic origin. This tumor expressed the fusion gene EWSR1-ATF1, which underlies the etiology of hyalinizing clear cell carcinoma (HCCC) and potentially that of CCOC. We found a breakpoint in the EWSR1-ATF1 fusion to be the same as that reported in HCCC. Established CCOC-T cells grew extremely slowly, but the cells showed highly invasive activity. Moreover, CCOC-T cells expressed bone-related molecules, odontogenic factors, and epithelial mesenchymal transition (EMT)-related molecules. Conclusion: To the best of our knowledge, this is the first report on the establishment of a CCOC cell line. CCOC-T cells serve as a useful in vitro model for understanding the pathogenesis and nature of MOT.

Oral cancer cells have a significantly augmented nuclear factor-κB (NF-κB) activity and the inhibition of this activity suppresses tumor growth. Bortezomib is a proteasome inhibitor and a drug used for molecular-targeted therapy (targets NF-κB). In this study, we investigated whether bortezomib would be effective as an inhibitor of proliferation and a radiosensitizer for the treatment of oral cancer. We demonstrate that bortezomib inhibits NF-κB activity and cell proliferation. The combined treatment with bortezomib and radiation (RT) suppressed NF-κB activity and cell growth in vitro and in vivo compared with RT treatment alone. To investigate the mechanisms by which bortezomib suppresses tumor growth, the expression of signaling molecules downstream of NF-κB were examined by ELISA. The combined treatment significantly inhibited the radiation-induced production of angiogenic factors and decreased the number of blood vessels in the tumor tissues. Although the expression of anti-apoptotic proteins was upregulated by RT, bortezomib downregulated the RT-induced expression of these proteins. Moreover, the expression of cleaved poly(ADP-ribose) polymerase in vitro and in vivo was enhanced by bortezomib, indicating that bortezomib inhibits tumor growth by inducing apoptosis. This study clearly demonstrates that bortezomib significantly inhibits tumor growth and that the combined treatment with bortezomib and RT results in a significant inhibition of tumor growth. The mechanisms underlying the inhibition of tumor growth by bortezomib include the suppression of angiogenesis and the induction of apoptosis. A novel molecular targeting therapy including bortezomib may be effective in the treatment of oral cancer by suppressing NF-κB activity.

Carbonate apatite (CO3Ap) foam has gained much attention in recent years because of its ability to rapidly replace bone. However, its mechanical strength is extremely low for clinical use. In this study, to understand the potential of gelatin-reinforced CO3Ap foam for bone replacement, CO3Ap foam was reinforced with gelatin and the resulting physical characteristics were evaluated. The mechanical strength increased significantly with the gelatin reinforcement. The compressive strength of gelatin-free CO3Ap foam was 74 kPa whereas that of the gelatin-reinforced CO3Ap foam, fabricated using 30 mass % gelatin solution, was approximately 3 MPa. Heat treatment for crosslinking gelatin had little effect on the mechanical strength of the foam. The gelatin-reinforced foam did not maintain its shape when immersed in a saline solution as this promoted swelling of the gelatin; however, in the same conditions, the heat-treated gelatin-reinforced foam proved to be stable. It is concluded, therefore, that heat treatment is the key to the fabrication of stable gelatin-reinforced CO3Ap foam.

Background The purpose of this retrospective study was to compare fluorine-18 fluorodeoxyglucose positron emission tomography/computed tomography (PET/CT) and ultrasonography (US) in the staging of patients with squamous cell carcinoma of the oral cavity. Methods We compared preoperative evaluations regarding lymph nodes using PET/CT, US, and both methods. The cutoff for the maximum standardized uptake value (SUVmax) in PET/CT was set at 2.7 by a receiver operating characteristic analysis that was based on the histopathological diagnosis. US was used to examine internal structural changes on B-mode and hilar vascularity on power Doppler. Results The performance of PET/CT and US in combination was better than that of each modality separately. However, there were histopathological changes that could not be detected on PET/CT or US. PET/CT could not detect nodes with necrotic or cystic changes. US could not detect lymph nodes that did not have abnormal structures. Conclusions PET/CT and US are complementary tools to evaluate preoperative patients.

Salivary gland cancers (SGCs) frequently metastasize to cervical lymph nodes and distant organs. Currently, the mechanisms responsible for the metastatic behavior of SGC cells are not fully understood. We previously demonstrated that the stromal cell-derived factor-1 (SDF-1; also known as CXCL12)/CXCR4 system is involved in the establishment of metastasis in oral squamous cell carcinoma. In the present study, we investigated the role of CXCR4 in the metastatic behavior of SGCs. We examined the expression of CXCR4 mRNA and protein in human SGC cell lines by quantitative RT-PCR and western blotting, respectively. The expression of CXCR4 mRNA and protein were frequently upregulated in 5 out of 6 SGC cell lines. Functional CXCR4 expression was demonstrated by the ability of these SGC cell lines to migrate toward an SDF-1 gradient. SDF-1 rapidly activated extracellular signal-regulated kinase (ERK)1/2 in SGC cell lines. Immunohistochemical analysis revealed that CXCR4 protein expression was detected in either the nucleus or cytoplasm of cancer cells in 16 out of 20 tissues of adenoid cystic carcinoma (ACC) and in 4 out of 6 tissues of mucoepidermoid carcinoma, which are representative of SGC. Furthermore, ACC cell lines exhibited dramatic metastasis to the lung following intravenous inoculation, whereas AMD3100, a CXCR4 antagonist, significantly inhibited lung metastasis of the cells, ameliorated body weight loss and improved the survival rate of tumor-bearing nude mice. These results indicate that CXCR4 expression contributes to the metastatic potential of SGCs.

Docetaxel (DOC) and 5-fluorouracil (5-FU) are important anticancer agents widely used in the treatment of a variety of cancers including oral squamous cell carcinoma (OSCC). The purpose of this study was to determine the antitumor efficacy of the sequential administration of DOC and 5-FU against OSCC cells (B88 and CAL27 cells) in vitro and in vivo. In in vitro growth inhibition assays, sequential treatment with DOC followed by 5-FU was more effective in inhibiting cancer cell growth than 5-FU followed by DOC, single treatment with DOC or 5-FU, or combined treatment with DOC and 5-FU. Furthermore, DOC followed by 5-FU significantly inhibited tumor growth in vivo compared to 5-FU followed by DOC. To understand the mechanisms underlying the enhanced growth inhibitory effect of the administration sequence, DOC followed by 5-FU, we examined the expression of 5-FU metabolic enzymes such as thymidylate synthase (TS), dihydropyrimidine dehydrogenase (DPD) and orotate phosphoribosyl transferase (OPRT), which were known to regulate the antitumor effect of 5-FU, by real-time RT-PCR and western blot analysis. Downregulation of TS and DPD expression and upregulation of OPRT expression were induced by DOC treatment, suggesting that DOC enhanced the efficacy of 5-FU by altering the expression of its metabolic enzymes. These results indicate that sequential treatment with DOC followed by 5-FU could be a promising therapeutic strategy for oral cancer.

Despite the clinical adoption of distraction osteogenesis (DO), studies examining the bone healing process at the distraction gap in osteoporotic bone are limited. We examined the effect of osteoporosis in the ovariectomized rat on DO.Mid-diaphyseal osteotomies were performed on the femurs of ovariectomized (OVX) rats. External distractors were placed on these rats and also on sham-ovariectomized rats. After a 7-day latency period, distraction was carried out at a rate of 0.5mm/day for 10 days. The bone volume (BV) of the distraction gap was measured by Micro-focused X-ray computed tomography (micro-CT) at 0, 2, and 4 weeks after completion of the distraction, and the distraction gap was examined histologically.The BV of the distraction gap in the OVX group was significantly lower than that in the sham group at 2 and 4 weeks after completion of distraction (p0.01). On histological examination, the distraction gap in the OVX group was filled with scattered smaller bone trabeculae than those seen in the sham group at 4 weeks after completion of distraction. Osteoclast numbers at the distraction gap in the OVX group were significantly increased when compared to the sham group (p0.01).Bone turnover with osteoclast predominance in ovariectomized rats is likely to be the cause of a reduction in new bone formation at the distraction gap.

Three commercially available artificial bone substitutes with different compositions, hydroxyapatite (HAp, Neobone&reg, ), carbonate apatite (CO3Ap, Cytrans&reg, ), and &beta, tricalcium phosphate (&beta, TCP, Cerasorb&reg, ), were compared with respect to their physical properties and tissue response to bone, using hybrid dogs. Both Neobone&reg, (HAp) and Cerasorb&reg, (&beta, TCP) were porous, whereas Cytrans&reg, (CO3Ap) was dense. Crystallite size and specific surface area (SSA) of Neobone&reg, (HAp), Cytrans&reg, (CO3Ap), and Cerasorb&reg, TCP) were 75.4 &plusmn, 0.9 nm, 30.8 &plusmn, 0.8 nm, and 78.5 &plusmn, 7.5 nm, and 0.06 m2/g, 18.2 m2/g, and 1.0 m2/g, respectively. These values are consistent with the fact that both Neobone&reg, TCP) are sintered ceramics, whereas Cytrans&reg, (CO3Ap) is fabricated in aqueous solution. Dissolution in pH 5.3 solution mimicking Howship&rsquo, s lacunae was fastest in CO3Ap (Cytrans&reg, ), whereas dissolution in pH 7.3 physiological solution was fastest in &beta, TCP (Cerasorb&reg, ). These results indicated that CO3Ap is stable under physiological conditions and is resorbed at Howship&rsquo, s lacunae. Histological evaluation using hybrid dog mandible bone defect model revealed that new bone was formed from existing bone to the center of the bone defect when reconstructed with CO3Ap (Cytrans&reg, ) at week 4. The amount of bone increased at week 12, and resorption of the CO3Ap (Cytrans&reg, ) was confirmed. &beta, ) showed limited bone formation at week 4. However, a larger amount of bone was observed at week 12. Among these three bone substitutes, CO3Ap (Cytrans&reg, ) demonstrated the highest level of new bone formation. These results indicate the possibility that bone substitutes with compositions similar to that of bone may have properties similar to those of bone.

[Background] Cancer stem cells (CSCs) exhibit self-replication, self-differentiation, drug resistance, and immune evasion activities. Recently, CSC has become increasingly important in the treatment of malignant tumors. Cancer stem cells express specific molecules termed CSC marker, including sex determining region Y-box2 (SOX2), and octamer-binding transcription factor 4 (Oct4), and their expression has been reported to be the potential prognostic values. However, the prognostic values of SOX2 and Oct4 expression in patients with stage I and II oral cancer are less understood. [Purpose] The aims of present study were to evaluate the expression of SOX2 and Oct4 in stage I and II oral squamous cell carcinoma (OSCC) and to elucidate the relationships among the CSC marker expression, clinical stages, histological differentiation, classification of invasion mode, cerebral lymph node metastasis, distant metastasis, and disease-free survival rate. [Materials and Methods] Tissue specimens were obtains from 83 patients with OSCC after surgery or biopsy. Immunohistochemistry was used to assess SOX2 and Oct4 protein using at least 10% staining-positive cells as the definition of positive staining. [Results] Immunohistochemical analysis of 83 cases showed that 31 cases (37%) expressed SOX2. There was no significant association between SOX2 expression and tumor size, invasion mode or histological differentiation. However, there was significant association between SOX2 expression and distant metastasis or disease-free survival rate at stage 1 and 2 patients. Otherwise, fifty three cases (64%) of 83 OSCC patients expressed Oct4. There was significant association between Oct4 expression and histological differentiation. There was no significant association between Oct4 expression and tumor size, invasion mode, metastasis, or disease-free survival rate. [Conclusions] These findings suggested that the expression of SOX2 may be good marker indicating survival in patients with stage 1 and 2 OSCC. Citation Format: Tetsuya Tamatani, Natsumi Takamaru, Go Ohe, Takayuki Nakagawa, Naito Kurio, Keiko Kudou, Yoshiko Yamamura, Takamitsu Mano, Youji Miyamoto. The clinicopathologic roles of SOX2 and Oct4 expressions in stage 1 and 2 oral squamous cell carcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 3061.

OBJECTIVES: Nasopharyngeal tonsilloliths (NT) and eustachian tube tonsilloliths (ET) are not as well-known to radiologists as palatine and lingual tonsilloliths. The aim of this investigation was to determine the prevalence and imaging characteristics of NT and ET using CT and panoramic radiographs. METHODS: We retrospectively assessed the scans of 2244 patients who underwent consecutive CT and panoramic radiographs of the maxillofacial region. The prevalence, size, number, and position of NT and ET were analysed. RESULTS: NT and ET were detected in 14 (0.6%) and 6 (0.3%) of 2244 patients on CT, respectively, but they were undetectable on panoramic radiographs. No significant difference was found in the prevalence with respect to sex. Although there was also no significant difference in the prevalence among age groups, tonsilloliths were most commonly noted in patients over 40 years old; they appeared as small and round calcified bodies, ranging from 1 to 3 mm in diameter. All NT were found 0 to 3 mm beneath the nasopharyngeal mucosal surface. CONCLUSIONS: The prevalence of NT and ET on CT was lower than that of palatine and lingual tonsilloliths. However, since they are encountered more frequently than clinically significant calcifying diseases such as retropharyngeal calcific tendinitis, clinicians should be able to correctly diagnose NT and ET based on their anatomical features.

Low crystalline B-type carbonate apatite (CO3Ap) block was fabricated by compositional transformation based on dissolution–precipitation reaction using microporous low crystalline calcite block as a precursor. In other words, microporous low crystalline calcite block prepared by carbonation of Ca(OH)2 compact was phosphoridated by exposing to 1 mol·dm−3 Na2HPO4 solutions at 60°C for 14 days. XRD, FT-IR and SEM analysis showed that calcite block completely transformed into B-type CO3Ap keeping its macroscopic morphology. Diametral tensile strength of the low-crystalline B-type CO3Ap block was approximately 5.4 MPa, and this value was much higher when compared with the low-crystalline calcite block, 0.73 MPa. Low crystalline B-type CO3Ap block fabricated in this method could be an ideal bone replacement due to its close similarity in its inorganic chemical composition and crystallinity to bone.