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Bilobalide isolated from the leaves of Ginkgo biloba has several pharmacological activities such as neuroprotective, anti-inflammatory, and anticonvulsant. However, the effect of bilobalide on cancer has not been clearly established. The main purpose of this study was to investigate the effect of bilobalide on cell growth and apoptosis induction in FaDu human pharyngeal squamous cell carcinoma. This was examined by 3-[4,5-dimethylthiazol-2-yl]-2,5- diphenyltetrazolium bromide assay, nuclear 4′,6-diamidino-2-phenylindole dihydrochloride staining, DNA fragmentation analysis, and immunoblotting. Bilobalide inhibited the growth of FaDu cells in dose- and time-dependent manners. Treatment with bilobalide resulted in nuclear condensation and DNA fragmentation in FaDu cells. Furthermore, it promoted the proteolytic cleavage of procaspase-3/-7/-8/-9 with increase in the amount of cleaved caspase-3/-7/-8/-9. Bilobalide-induced apoptosis in FaDu cells was mediated by the expression of Fas and the activation of caspase-8, caspase-3, and poly (ADP-ribose) polymerase. Immunoblotting revealed that the antiapoptotic mitochondrial protein Bcl-2 was downregulated, but the proapoptotic protein Bax was upregulated by bilobalide in FaDu cells. Bilobalide significantly increased Bax/Bcl-2 ratio. These results suggest that bilobalide inhibits cell proliferation and induces apoptosis in FaDu human pharyngeal squamous cell carcinoma via both the death receptor-mediated extrinsic apoptotic pathway and the mitochondrial-mediated intrinsic apoptotic pathway.

Purpose: This study examines the properties and potential commercial value of the pine cones extract when used as bioactive and cosmetics ingredients. Methods: Pine cones were extracted with hot water (WE), 70% ethanol (EE), and supercritical fluid extraction (SFE) to examine their antioxidant properties on the basis of their total polyphenol content, 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity, and gas chromatography-mass spectrometry (GC-MS) analysis. The antibacterial properties of the pine cone extract were also studied on 2 types of bacteria. In addition, this study examined the nitric oxide (NO) inhibition effect using RAW 264.7 cells, in which the inflammatory reaction was induced by lipopolysaccharides (LPS), and cell migration activity in the HeCaT cells. Results: The extract obtained with SFE exhibited high antioxidative ability even at low concentration (0.125%). The antimicrobial activity of the of SFE pine cone extract was 12.1±1.04 mm for Staphylococcus aureus (S. aureus ) and 11.8±0.45 mm for Staphylococcus epidermidis (S. epidermidis ). Cytotoxicity evaluation showed cell viability above 80% in all extracts. During the antiinflammatory test, the nitric oxide expression level was low, and it was confirmed by the antiinflammatory effect. The results of the stability test showed that emulsions containing SFE extract were very stable. The pH and high temperature (40℃) of each emulsion containing SFE extract did not considerably change for 30 days. Conclusion: These findings suggest that SFE extract is a possible cosmeceutical material with potent skin regeneration and antiinflammatory properties.

Purpose The purpose of this study was to evaluate the effects of the growth factor within platelet-rich fibrin (PRF) in proliferation and differentiation of osteoblast and to observe the effectiveness of PRF. Materials and methods The colorimetric MTT assay, cell live and dead assay, alkaline phosphatase staining and activity assay, alizarine red S, and von Kossa staining were performed. Finally, the alterations of biomarkers associated with bone formation were verified at the mRNA level by quantitative polymerase chain reaction (PCR) and quantitative real-time PCR. In in vivo study, 6 adult mongrel dogs were used. The defect was performed and divided into 3 groups: (1) defect left unfilled, (2) defect filled with only 0.25-g Bio-Oss, and (3) defect filled with 0.25-g Bio-Oss mixed with PRF. Results MTT and cell live and dead assay showed that PRF did not affect the cell viability in MG-63 cells. The alkaline phosphatase activity, calcification, and mineralization were gradually increased in the MG-63 cells treated with PRF. Furthermore, the mRNA levels of biomarker gene in the MG-63 cells treated with PRF were significantly higher than those of control. In in vivo study, both radiographical and histological evaluations showed that the new bone formations were significantly increased in the defecting bone region transplanted with Bio-Oss and PRF compared with Bio-Oss only at 2 weeks after transplantation. Conclusion PRF can promote the bone regeneration without any complications.

Purpose The purpose of this study was to evaluate a new graft material, biphasic calcium phosphate, composed of 60% hydroxyapatite and 40% β-Tricalcium phosphate and deproteinized bovine bone mineral, which is established as a predictable graft material for maxillary sinus augmentation. Materials and methods Maxillary sinus augmentation was performed with different bone materials. Bone biopsies were performed on tissue harvested from the future implant bed using a trephine bur at 6 months after maxillary sinus augmentation. Resonance frequency analysis was performed immediately and at 6 months after the implant placement. Microcomputed tomography and histomorphometric analysis were performed in all patients. Results Fifty-six patients (60 sinuses) were included in the study. At 6 months postoperative, 31 biopsies were performed on tissues harvested from the calcium phosphate, and 29 biopsies on tissues from the bovine bone grafts. There was no implant failure during the 21-month mean follow-up period. The overall implant stability quotient values were higher than 60, and gradually increased for 6 months. Higher new bone volume fraction and new bone surface density were observed in the calcium phosphate group compared with the bovine bone group. In contrast, residual bone graft volume in the bovine bone group was higher than that in the calcium phosphate group. Nevertheless, there was no significant difference between groups in the microcomputed tomography and histomorphometric parameters. Conclusion Within the study's limitations, both graft materials demonstrated similar biocompatibility and osteoconductivity in the maxillary sinus augmentation.

Background/purpose: The classification and treatment of odontogenic keratocyst (OKC) are controversial. The objective of this study was to present the efficiency of decompression followed by enucleation by clinical and histomorphometric evaluation for the treatment of OKC. Materials and methods: Thirty four OKCs of 27 patients who underwent decompression followed by enucleation were included in this study. Clinical and histomorphometric analysis were performed. Results: The average decreasing rate was 59% in maximum diameter, 66% in the amount of the volume for the average of period of the decompression was 9.8 months. The mean of increasing rate of the thickness of the epithelial lining was 921.16%. There were no recurrences for a mean follow-up period of 5.8 years. The thin and friable cyst wall of the OKC was changed to thickened, hard type. Conclusion: The decompression was found to be effective and reliable as a treatment of the OKC to decrease the recurrence tendency, even for Gorlin-Goltz syndrome. Keywords: Epithelial thickness, Jaw cysts, Keratocysts, Odontogenic tumors

Background/aim MicroRNAs (miRNAs) are closely associated with a number of cellular processes, including cell development, differentiation, proliferation, carcinogenesis, and apoptosis. The aim of the present study was to elucidate the molecular mechanisms underlying the tumor suppressor activity of miRNA-203 (miR-203) in YD-38 human oral cancer cells. Materials and methods Polymerase chain reaction analysis, MTT assay, DNA fragmentation assay, fluorescence-activated cell-sorting analysis, gene array, immunoblotting, and luciferase assay were carried out in YD-38 cells. Results miR-203 expression was significantly down-regulated in YD-38 cells compared to expression levels in normal human oral keratinocytes. miR-203 decreased the viability of YD-38 cells in a time- and dose-dependent manner. In addition, over-expression of miR-203 significantly increased not only DNA segmentation, but also the apoptotic population of YD-38 cells. These results indicate that miR-203 overexpression induces apoptosis in YD-38 cells. Target gene array analysis revealed that the expression of the polycomb complex protein gene Bmi-1, a representative oncogene, was significantly down-regulated by miR-203 in YD-38 cells. Moreover, both mRNA and protein levels of Bmi-1 were significantly reduced in YD-38 cells transfected with miR-203. These results indicate that Bmi-1 is a target gene of miR-203. A luciferase reporter assay confirmed that miR-203 suppressed Bmi-1 expression by directly targeting the 3'-untranslated region. Conclusion miR-203 induces apoptosis in YD-38 cells by directly targeting Bmi-1, which suggests its possible application as an anti-cancer therapeutic.

The fracture of dental implants is a rare occurrence in clinical settings. Possible causes of implant fracture include design or production flaws, overloaded occlusion force, implant location, metal fatigue, and bone resorption around the implant. This study reports on the successful removal and reimplantation of fractured implants.

Metformin (1,1-dimethylbiguanide hydrochloride), derived from French lilac (Galega officinalis), is a first-line drug prescribed for patients with type 2 diabetes. It has been reported to have anti-cancer effects in a variety of cancer cells. However, effects of metformin on oral cancer cells have not been clearly established. The main goal of this study was to investigate the effect of metformin on cell growth and apoptosis induction in oral cancer cells derived from Korean patients. The effect of metformin on cell growth and apoptosis induction in oral cancer cells was examined by inhibition of cell growth, DNA fragmentation analysis and immunoblotting in YD-38 human oral cancer cells derived from Korean patients. Treatment with metformin induced inhibition of cell growth depending on the metformin treatment time and concentration in YD-38 human oral cancer cells. Treatment with metformin induced nuclear fragmentation in YD-38 human oral cancer cells. Metformin promoted proteolytic cleavage of procaspase-3 with an increase in the amount of cleaved caspase-3. Cleaved PARP was increased by metformin in YD-38 human oral cancer cells. Treatment of YD-38 human oral cancer cells with metformin increased the level of Bax, but it decreased the level of Bcl-2. These results suggest that metformin can induce suppression of cell growth and cell apoptosis in YD-38 human oral cancer cells derived from Korean patients.

Background/aim The present study aimed to investigate the apoptotic effects of phenformin, a therapeutic agent for diabetes, on head and neck squamous cell carcinoma (HNSCC). Materials and methods Cytotoxicity was measured by the MTT and live/dead cell assay. Phenformin-induced apoptotic FaDu cell death and its associated cellular signaling pathways were investigated by hematoxylin and eosin staining, 4',6-diamidino-2-phenylindole staining, caspase-3 activity assay, fluorescence-activated cell sorting analysis, and western blotting. Results Phenformin promoted death of and apoptotic processes in FaDu cells, including morphological alterations and nuclear condensation. Furthermore, treatment with phenformin increased caspase-3 activity and apoptotic populations via the caspase cascade through cleavage of capspase-8, -9, and -3 and poly(ADP-ribose) polymerase in FaDu cells. Moreover, phosphorylation levels of mitogen-activated protein kinases, nuclear factor-κB, and AKT were down-regulated in FaDu cells by phenformin. Conclusion Phenformin induced death of FaDu cells via caspase-dependent extrinsic and intrinsic apoptosis pathways and is a promising novel therapeutic agent for HNSCC.

In the present study, we demonstrated the anti-catabolic effects of formononetin, a phytoestrogen derived from herbal plants, against interleukin-1β (IL-1β)-induced severe catabolic effects in primary rat chondrocytes and articular cartilage. Formononetin did not affect the viability of primary rat chondrocytes in both short- (24 h) and long-term (21 days) treatment periods. Furthermore, formononetin effectively antagonized the IL-1β-induced catabolic effects including the decrease in proteoglycan content, suppression of pericellular matrix formation, and loss of proteoglycan through the decreased expression of cartilage-degrading enzymes like matrix metalloproteinase (MMP)-13, MMP-1, and MMP-3 in primary rat chondrocytes. Moreover, catabolic oxidative stress mediators like nitric oxide, inducible nitric oxide synthase, cyclooxygenase-2, and prostaglandin E2 were significantly downregulated by formononetin in primary rat chondrocytes treated with IL-1β. Sequentially, the upregulation of pro-inflammatory cytokines (like IL-1α, IL-1β, IL-6, and tumor necrosis factor α), chemokines (like fractalkine, monocyte chemoattractant protein-1, and macrophage inflammatory protein-3α), and vascular endothelial growth factor were significantly downregulated by formononetin in primary rat chondrocytes treated with IL-1β. These data suggest that formononetin may suppress IL-1β-induced severe catabolic effects and osteoarthritic condition. Furthermore, formononetin may be a promising candidate for the treatment and prevention of osteoarthritis.

Morin, a flavonoid isolated from various medicinal herbal plants, has an anti-inflammatory effect. This study aimed to elucidate the anticatabolic effects and cellular mechanism of morin against interleukin-1β (IL-1β) in rat primary chondrocytes. Morin at 10–100 μM did not affect the viability of rat primary chondrocytes. Treatment with morin for 21 days ameliorated the IL-1β-induced decrease in extracellular matrix. Furthermore, treatment with morin attenuated IL-1β-induced proteoglycan loss in the articular cartilage through suppression of catabolic factors, such as matrix metalloproteinases, inflammatory mediators, and pro-inflammatory cytokines. These data indicated that morin exerted anticatabolic effects that can prevent and reduce progressive degeneration of the articular cartilage, and thus may be a potential candidate treatment for osteoarthritis.

In the present study, we investigated the anti-catabolic effects of coumestrol, a phytoestrogen derived from herbal plants, against interleukin-1β-induced cartilage degeneration in primary rat chondrocytes and articular cartilage. Coumestrol did not affect the viability of human normal oral keratinocytes and primary rat chondrocytes treated for 24 h and 21 days, respectively. Although coumestrol did not significantly increase the proteoglycan contents in long-term culture, it abolished the interleukin-1β-induced loss of proteoglycans in primary rat chondrocytes and knee articular cartilage. Furthermore, coumestrol suppressed the expression of matrix-degrading enzymes such as matrix metalloproteinase-13, −3, and −1 in primary rat chondrocytes stimulated with interleukin-1β. Moreover, the expression of catabolic factors such as nitric oxide synthase, cyclooxygenase-2, prostaglandin E2, and inflammatory cytokines in interleukin-1β-stimulated primary rat chondrocytes was suppressed by coumestrol. In summary, these results indicate that coumestrol counteracts the catabolic effects induced by interleukin-1β through the suppression of inflammation. Therefore, based on its biological activity and safety profile, coumestrol could be used as a potential anti-catabolic biomaterial for osteoarthritis.

Implant displacement into the maxillary sinus often results from features specific to the posterior maxillary teeth, including poor bone quality and insufficient remaining bone. This study reviews implants displaced into the maxillary sinus, the causes and complications of displacement, and how to remove them, according to when the displacement occurs.The PubMed, Ovid (MEDLINE), and EMBASE databases were searched using the keywords "displacement," "implant," "maxillary sinus," and "removal" for articles published between January 2000 and July 2013.Twenty-two journal articles were selected; these discussed 49 displaced implants. Most of the implants were displaced into the maxillary sinus during implantation, but resulted in a low incidence of complications, such as maxillary sinusitis. The displaced implants were removed using the Caldwell-Luc approach or a transoral or transnasal endoscopic approach.Implants displaced into the maxillary sinus have various causes according to when they are displaced. As displaced implants can cause several complications, transnasal endoscopy is recommended to remove them; however, the implants should be examined thoroughly before selecting the removal method.

This analysis examined the types of retreatment in failed sinus bone grafts due to the development of maxillary sinusitis. Reoperation was performed in 7 patients. The types of reoperation included infection management, reconstruction of the sinus roof using a pedicled buccal fat pad and collagen membrane, oroantral fistula closure, sinus bone graft using an autogenous bone graft, and implant placement. In one case, sinusitis developed 14 months after the reoperation, but it was managed by incision, drainage, and administration of antibiotics. All sinus bone grafts that were performed during the retreatments were successful.

MicroRNAs (miRNAs) are closely associated with a number of cellular processes, including cell development, differentiation, proliferation, carcinogenesis, and apoptosis. The aim of the present study was to elucidate the molecular mechanisms underlying the tumor suppressor activity of miRNA-203 (miR-203) in YD-38 human oral cancer cells.Polymerase chain reaction analysis, MTT assay, DNA fragmentation assay, fluorescence-activated cell-sorting analysis, gene array, immunoblotting, and luciferase assay were carried out in YD-38 cells.miR-203 expression was significantly down-regulated in YD-38 cells compared to expression levels in normal human oral keratinocytes. miR-203 decreased the viability of YD-38 cells in a time- and dose-dependent manner. In addition, over-expression of miR-203 significantly increased not only DNA segmentation, but also the apoptotic population of YD-38 cells. These results indicate that miR-203 overexpression induces apoptosis in YD-38 cells. Target gene array analysis revealed that the expression of the polycomb complex protein gene Bmi-1, a representative oncogene, was significantly down-regulated by miR-203 in YD-38 cells. Moreover, both mRNA and protein levels of Bmi-1 were significantly reduced in YD-38 cells transfected with miR-203. These results indicate that Bmi-1 is a target gene of miR-203. A luciferase reporter assay confirmed that miR-203 suppressed Bmi-1 expression by directly targeting the 3'-untranslated region.miR-203 induces apoptosis in YD-38 cells by directly targeting Bmi-1, which suggests its possible application as an anti-cancer therapeutic.

Background Socket sealing surgery is performed for the preservation of the form and volume of the soft tissue by covering the resulting socket with autogenous soft tissue graft or membrane barriers. This procedure is usually necessary to improve the esthetic results of the maxillary anterior or premolar areas. Methods This study retrospectively investigated cases involving the open membrane technique or socket sealing surgery with a palatal gingival graft or collagen membrane where implant placement and bone grafting were performed immediately after tooth extraction. From January 2005 to December 2008, socket sealing surgery was performed in 24 patients, and 25 implants were placed. Results All implants were successful in the follow-up period. In the palatal gingival graft group, the mean marginal bone loss was 1.17 mm during the mean follow-up period of 81.0 months. In the collagen membrane group, the mean marginal bone loss was 1.23 mm during the mean follow-up period of 76.9 months. There was no significant difference between the two groups. Conclusions Consequently, socket sealing surgery is effective at minimizing the loss of soft tissue and alveolar bone. Electronic supplementary material The online version of this article (10.1186/s40902-017-0137-x) contains supplementary material, which is available to authorized users.

In the present study, we investigated berberine‑induced apoptosis and the signaling pathways underlying its activity in FaDu head and neck squamous cell carcinoma cells. Berberine did not affect the viability of primary human normal oral keratinocytes. In contrast, the cytotoxicity of berberine was significantly increased in FaDu cells stimulated with berberine for 24 h. Furthermore, berberine increased nuclear condensation and apoptosis rates in FaDu cells than those in untreated control cells. Berberine also induced the upregulation of apoptotic ligands, such as FasL and TNF-related apoptosis-inducing ligand, and triggered the activation of caspase-8, -7 and -3, and poly(ADP ribose) polymerase, characteristic of death receptor-dependent extrinsic apoptosis. Moreover, berberine activated the mitochondria‑dependent apoptotic signaling pathway by upregulating pro-apoptotic factors, such as Bax, Bad, Apaf-1, and the active form of caspase-9, and downregulating anti-apoptotic factors, such as Bcl-2 and Bcl-xL. In addition, berberine increased the expression of the tumor suppressor p53 in FaDu cells. The pan-caspase inhibitor Z-VAD-fmk suppressed the activation of caspase-3 and prevented cytotoxicity in FaDu cells treated with berberine. Interestingly, berberine suppressed cell migration through downregulation of vascular endothelial growth factor (VEGF), matrix metalloproteinase (MMP)-2, and MMP-9. Moreover, the phosphorylation of extracellular signal-regulated kinase (ERK1/2) and p38, components of the mitogen-activated protein kinase pathway that are associated with the expression of MMP and VEGF, was suppressed in FaDu cells treated with berberine for 24 h. Therefore, these data suggested that berberine exerted anticancer effects in FaDu cells through induction of apoptosis and suppression of migration. Berberine may have potential applications as a chemotherapeutic agent for the management of head and neck squamous carcinoma.

We report generation and characterization of pain-related behavior in a minimally invasive facet joint degeneration (FJD) animal model in rats. FJD was produced by a non-open percutaneous puncture-induced injury on the right lumbar FJs at three consecutive levels. Pressure hyperalgesia in the lower back was assessed by measuring the vocalization response to pressure from a force transducer. After hyperalgesia was established, pathological changes in lumbar FJs and alterations of intervertebral foramen size were assessed by histological and imaging analyses. To investigate treatment options for lumber FJ osteoarthritis-induced pain, animals with established hyperalgesia were administered with analgesic drugs, such as morphine, a selective COX-2 inhibitor, a non-steroidal anti-inflammatory drug (NSAID) (ketorolac), or pregabalin. Effects were assessed by behavioral pain responses. One week after percutaneous puncture-induced injury of the lumbar FJs, ipsilateral primary pressure hyperalgesia developed and was maintained for at least 12 weeks without foraminal stenosis. Animals showed decreased spontaneous activity, but no secondary hyperalgesia in the hind paws. Histopathological and microfocus X-ray computed tomography analyses demonstrated that the percutaneous puncture injury resulted in osteoarthritis-like structural changes in the FJs cartilage and subchondral bone. Pressure hyperalgesia was completely reversed by morphine. The administration of celecoxib produced moderate pain reduction with no statistical significance while the administration of ketorolac and pregabalin produced no analgesic effect on FJ osteoarthritis-induced back pain. Our animal model of non-open percutanous puncture-induced injury of the lumbar FJs in rats shows similar characteristics of low back pain produced by human facet arthropathy.

Purpose: The aim of the present study is to assess the importance of proper treatment timing for dental emergency patients by characterizing current patient care in the emergency room. Materials and Methods: A retrospective chart review of 3,211 patients who visited the Chosun University Hospital’s dental emergency department (Gwangju, Korea) was conducted from January 1, 2011 to May 31, 2014. Information regarding age, gender, onset date, main causes, and diagnoses were collected and analyzed. The main causes were divided into six categories: assault, household/play, sports, traffic, work, and others. Result: Emergency visits were more common for men (69%), and the ratio of males to females was 2.2:1 On average, the major cause was household/play (49.8%), followed by others (18.9%), traffic (16.6%), assault (9.1%), sports (2.9%), and work (2.6%). The most frequent diagnosis on average was dental trauma with 82.4%, followed by infection (10.7%), others (4.7%), and bleeding (2.2%). Conclusion: The main reasons for visits to the dental emergency department are dental trauma, dental infection, bleeding, and others. The most frequent reason for dental emergency patients to visit the emergency department was dental trauma (82.4%).

The purpose of this study was to compare different internal fixation techniques with resorbable or titanium fixation system used in sagittal split ramus osteotomy (SSRO) for mandibular setback. Synthetic polyurethane hemimandible replicas were used. The distal segment was repositioned in a 5-mm setback position. The hemimandibles were divided into the following eight groups: resorbable miniplate (A), titanium miniplate (B), resorbable hybrid; resorbable miniplate and bicortical screw (C), titanium hybrid; titanium miniplate and bicortical screw (D), resorbable mixed hybrid; resorbable bicortical screw and titanium miniplate (E), titanium mixed hybrid; titanium bicortical screw and resorbable miniplate (F), three resorbable bicortical screws (G) and three titanium bicortical screws (H). The compression loads were applied on the incisal edge. The compression loads (N) at 5- and 10-mm displacement were analyzed (P < 0.05). Miniplate groups (A and B) were significantly weaker than the other groups (C ∼ H). Group (H) had greatest biomechanical stability. There was no significant difference between group (D) and group (H) at 10-mm displacement. Group (G) showed a lower load than group (H) and group (D). There was no significant difference between the hybrid and mixed hybrid groups. It was concluded that additional placement of a bicortical screw, either titanium or resorbable with a miniplate may provide significantly better stabilization.

We investigated Licochalcone-A (Lico-A)-induced apoptosis and the pathway underlying its activity in a pharyngeal squamous carcinoma FaDu cell line. Lico-A purified from root of Glycyrrhiza inflata had cytotoxic effects, significantly increasing cell death in FaDu cells. Using a cell viability assay, we determined that the IC50 value of Lico-A in FaDu cells was approximately 100 µM. Chromatin condensation was observed in FaDu cells treated with Lico-A for 24 h. Consistent with this finding, the number of apoptotic cells increased in a time-dependent manner when FaDu cells were treated with Lico-A. TRAIL was significantly up-regulated in Lico-A-treated FaDu cells in a dose-dependent manner. Apoptotic factors such as caspases and PARP polymerase were subsequently activated in a caspase-dependent manner. In addition, levels of pro-apoptotic factors increased significantly in response to Lico-A treatment, while levels of anti-apoptotic factors decreased. Lico-A-induced TRAIL expression was mediated in part by a MAPK signaling pathway involving ERK1/2 and p38. Lastly, in an in vivo xenograft mouse model, Lico-A treatment effectively suppressed the growth of FaDu cell xenografts by activating caspase-3, without affecting the body weight of mice. Taken together, these data suggest that Lico-A has potential chemopreventive effects and should therefore be developed as a chemotherapeutic agent for pharyngeal squamous carcinoma.

The neurotrophin-inducible gene VGF plays an important role in the maintenance of organismal energy balance and in the mediation of hippocampal synaptic activity. The regulatory mechanism of VGF transcription is not fully understood. The neuron-restrictive silencer factor (NRSF) binds with the neuron-restrictive silencer element (NRSE), thereby suppressing the transcription of NRSE-containing genes. In this study, we show that the NRSE sequence of the VGF gene critically regulates the repression of VGF expression in NMB cells. Sequence analysis also establishes the presence of two putative NRSEs (NRSE-1 and NRSE-2) in the promoter region of the VGF gene. In reporter gene experiments, a more than eight-fold increase in the promoter activity was observed when both NRSE-1 and NRSE-2 were deleted. Deletion of NRSE-2 alone did not affect the promoter activity, thus indicating that NRSE-1 could be solely responsible for the repression of VGF gene expression. Mutations in the NRSE-1 sequence increased promoter activity. However, no change in activity was observed when NRSE-1 was coexpressed with dominant-negative NRSF, thereby suggesting that endogenous NRSF interacts with NRSE-1. Binding of NRSF to NRSE in a sequence-specific manner was confirmed with chromatin immunoprecipitation assays, respectively. Furthermore, the overexpressed NRSF in PC12 cells significantly suppressed the VGF gene expression by interacting with the NRSE located in the VGF promoter region. Our results indicate that NRSF plays an important role as a repressor of VGF gene regulation in NMB cells through a mechanism that is dependent on VGF-NRSE.

PURPOSE The purpose of this study was to evaluate the ability of new bone formation of deproteinized bovine bone (Bio-Oss) and mineralized allogenic bone (Tutoplast). MATERIALS AND METHODS Sixty rats were divided into control and experimental groups (groups 1 and 2): control group, unfilled control; group 1, Bio-Oss; group 2, Tutoplast, respectively. The animals were killed after 6 and 12 weeks, and newly formed bone was analyzed histomorphometrically. RESULTS In the control group, some new bone formed in the rim of the defect area. In the group 1, newly formed bone was thinner than the adjacent normal bone, and Bio-Oss particles were observed. In the group 2, showed a pattern of gradual fusion with adjacent bone, as well as particles in some areas, similar to the Bio-Oss-treated group. In the 12-week groups, the amount of new bone formation was significantly higher in the experimental groups than in the control group, and it was significantly higher in group 2 than in group 1. CONCLUSION Although Tutoplast and Bio-Oss graft materials seem to be useful for bone grafts, Tutoplast showed more active new bone formation than Bio-Oss.

PURPOSE This study evaluated the effect of drilling speed on early bone healing in the mandible of dogs. MATERIAL AND METHODS Six dogs were selected, and mandibular premolars and molars were extracted. After 2 months, 3 hydroxyapatite-surfaced fixtures were implanted with drilling speeds of 50, 800, and 1200 rpm on the right side first and then on the left side after 2 weeks. Implant stability quotient (ISQ) was measured on insertion, after 2 and 4 weeks. RESULTS Based on the ISQ measurement, the 1200-rpm group showed a higher value than the 50-rpm group at 2 weeks and 4 weeks (P < 0.05). New bone formation around the implant was highest for the 800-rpm group at 2 weeks and the 1200-rpm group at 4 weeks. The bone-implant contact of the superior half of the alveolar bone was highest for the 800-rpm group at 2 weeks and the 1200-rpm group at 4 weeks. There was no statistically significant difference. CONCLUSION This study suggests that 50, 800, and 1200 rpm are drilling speeds which can expect favorable outcome, yet, higher drilling speed presented overall the best biological responses.

BACKGROUND/AIM The purpose of this study was to elucidate the molecular mechanism underlying regulation of semaphorin-6A (SEMA6A) involving microRNA-203 (miR-203) as a tumor suppressor in YD-38 human oral cancer cells. MATERIALS AND METHODS miRNA arrays, polymerase chain reaction analyses, MTT assays, immunoblotting, and luciferase assays were carried out in YD-38 cells. RESULTS MiRNA microarray results showed that expression of miR-203 was significantly down-regulated in YD-38 cells compared to normal human oral keratinocytes. The viability of YD-38 cells was reduced by miR-203 in time- and dose-dependent manners. Overexpression of miR-203 increased the nuclear condensation of YD-38 cells and activated the apoptotic signaling pathway by up-regulating pro-apoptotic factors, such as BCL-2-associated X protein (BAX) and BCL-2 homologous antagonist killer (BAK), and the active forms of caspase-9, caspase-3, and poly-(ADP-ribose)-polymerase (PARP). Furthermore, target gene array analyses revealed that the expression of class 6 semaphorin A (SEMA6A) was down-regulated by miR-203 in YD-38 cells. Both the mRNA and protein levels of SEMA6A were reduced in YD-38 cells transfected with miR-203. Luciferase activity assay confirmed that miR-203 directly targets the SEMA6A 3'-untranslated region to suppress gene expression. CONCLUSION Our results indicate that miR-203 induces the apoptosis of YD-38 human oral cancer cells by directly targeting SEMA6A, suggesting its potential application in anticancer therapeutics.

Background Codium fragile (Suringar) Hariot has been used as a potential remedy in traditional medicine because of its anti-inflammatory and anti-oxidant effects. Osteoarthritis is a chronic progressive joint disease, characterized by complex mechanisms related to inflammation and degeneration of articular cartilage. In this study, we aimed to evaluate the cartilage protective effect of an aqueous extract of Codium fragile (AECF) using rat primary chondrocytes and the osteoarthritis animal model induced by destabilization of the medial meniscus (DMM). Methods In vitro, rat primary cultured chondrocytes were pre-treated with AECF (0.5, 1, and 2 mg/mL) for 1 h and then incubated with interleukin-1β (10 ng/mL) for 24 h. Nitrite production was detected by the Griess reagent. Alteration of the protein levels of iNOS, MMP-13, ADAMTS-4, ADAMTS-5, mitogen-activated protein kinases (MAPKs), and nuclear factor-κB (NF-κB) was detected by western blotting. In vivo, osteoarthritis was induced by DMM of Sprague Dawley (SD) rats. The rats subjected to destabilization of the medial meniscus (DMM) surgery were orally administered with AECF (50, 100, and 200 mg/kg bodyweight) or distilled water for 8 w. The severity of cartilage lesions was evaluated by safranin O staining and the Osteoarthritis Research Society International (OARSI) score. Results These results demonstrated that AECF significantly inhibited nitrite production and inhibited the levels of iNOS, MMP-13, ADAMTS-4, and ADAMTS-5 in interleukin-1β-induced rat primary cultured chondrocytes. Moreover, AECF suppressed interleukin-1β-induced NF-κB activation in the nucleus and phosphorylation of ERK1/2 and JNK in the cytosol. In vivo, the cartilage lesions in AECF‐treated osteoarthritis rats exhibited less proteoglycan loss and lower OARSI scores. Conclusions These results suggested that AECF is a potential therapeutic agent for the alleviation of osteoarthritis progression.

PURPOSE The objective of this study was to compare the implant stability and osseointegration of implants using a flap or flapless technique. MATERIAL AND METHODS Mandibular premolars and molars were extracted from both sides in 6 dogs. After 8 weeks, 4 fixtures were implanted using either a flap or flapless technique. Implant stability quotient was measured on insertion and at 2, 4, and 8 weeks later. The animals were killed while the tissues were histologically analyzed. RESULTS Implant stability increased for 8 weeks, and no statistically significant differences were observed between the surgical protocols. Bone-implant contact showed 60.27% ± 30.99% for flapless surgery and 59.73% ± 17.12% for flap surgery. And the results of new bone formation area from total area showed 56.07% ± 27.78% for flapless surgery and 57.00% ± 14.66% for flap surgery. There were no statistically significant differences. CONCLUSION This study showed no significant difference in implant stability as well as osseointegration regardless of flap or flapless technique.

The aim of the present study was to investigate biochanin-A-induced anticancer effects and their cellular signaling pathway in FaDu pharyngeal squamous carcinoma cells. Biochanin-A induced cell death through increased cytotoxicity of FaDu cells in a dose- and time-dependent manner. The number of cells with nucleus condensation and the apoptotic population were increased in the FaDu cells stimulated with biochanin-A for 24 h. Furthermore, extrinsic apoptotic factors such as FasL and their downstream target caspase-8 were increased and activated in the FaDu cells treated with biochanin-A in a dose-dependent manner. Moreover, biochanin-A decreased the expression of intrinsic anti-apoptotic factors such as Bcl-2 and Bcl-xL, and increased the level and activation of intrinsic apoptotic factors such as Bad and caspase-9. Finally, biochanin-A induced the activation of caspase-3 and Poly(ADP ribose) polymerase (PARP) in FaDu cells. Our results suggest that biochanin-A-induced apoptosis was mediated by death receptor mediated-extrinsic and mitochondria-dependent intrinsic apoptotic signaling pathways. Biochanin-A also inhibited wound healing migration and proliferation of FaDu cells via the downregulation and inactivation of matrix metalloproteinase-2 and -9 that are mediated by the suppression of p38, mitogen activated protein kinase (MAPK), NF-κB and Akt cellular signaling pathways. Therefore, these data suggest that the biochanin-A may act as a potential chemotherapeutic compound to treat head and neck cancer.

The purpose of this study was to evaluate the clinical outcomes of patients undergoing sinus membrane elevation by a minimally invasive crestal approach using a novel drilling system.From May 2008 to November 2009, 21 implants were placed in 19 patients (10 men and 9 women) ranging from 23 to 69 years of age (average of 49.5 years). Implants were placed in maxillary premolar and molar areas that demonstrated insufficient residual bone quality; maxillary sinus membrane elevation was performed using a crestal approach with the sinus crestal approach kit (Neobiotech, Seoul, Korea).There was no sinus perforation or osseointegration failure. The implant survival rate was 100%. The postsurgical, augmented volume of the alveolar height ranged from 2 to 9.2 mm (average of 5.81 ± 2.06 mm). Six months after maxillary sinus elevation, the bone reduction volume ranged from 0.06 to 1.42 mm (average of 0.6 ± 0.38 mm). At final F/U, the amount of bone-height reduction ranged from 0.06 to 2.60 mm (average of 0.82 ± 0.63 mm).Sinus membrane elevation by the crestal approach using special reamers is advantageous because of the noticeable reduction in the risk of perforation and the ability to perform the surgery rapidly.

Purpose To develop a methodology for the synthesis of β-tricalcium phosphate (β-TCP, Ca3(PO4)2) from the shell of Haliotis sp. (abalone shell) and to verify its characterization and biocompatibility. Materials and methods Calcium oxide (CaO) was synthesized from abalone shell by sintering and was suspended in distilled water to prepare calcium hydroxide (Ca(OH)2). For the synthesis of calcium carbonate (CaCO3), carbon dioxide was used to infuse Ca(OH)2 at pH 7.4. CaCO3 was reacted with phosphoric acid at pH 6.0 to obtain dicalcium phosphate (CaHPO4). Subsequently, β-TCP was synthesized by a chemical reaction between CaHPO4 and CaO at 950°C to 1100°C for 3 hours. Fourier transform infrared spectroscopy (FT-IR) and x-ray diffraction (XRD) was performed to verify the physiochemical characteristics of the composite synthesized from abalone shell. Results FT-IR and XRD results showed that β-TCP was successfully synthesized from abalone shell. The synthesized β-TCP did not affect cell viability of either normal human oral keratinocytes or osteoblastic MG-63 cells. These data indicate that β-TCP synthesized from abalone shell is biologically safe. Conclusions β-TCP (Ca3(PO4)2) synthesized from abalone shell can be used as a potential source of bone grafting material.

Purpose: To establish management protocol for mandibular angle fracture, we describe pertinent factors including cause, impacted third molar and recent treatment tendency. Methods: We examined the records of 62 patients who had unilateral mandibular angle fracture. Sixty patients who had open reduction surgery were examined at postoperative weeks 1, 4, 8, 12, and 28. Results: Left mandibular angle fracture is frequent in younger males. Presence of the mandibular third molar can increase fracture risk. Because of attached muscle, favorable fractures occurred primarily in the mandibular angle area. Conclusion: Extracting the mandibular third molar can prevent angle fractures, and open reduction with only one plate adaptation is generally the proper treatment method for mandibular angle fracture.

Sambucus sieboldiana (SS) is a member of the family Caprifoliaceae and has been recommended as a functional material because of its several bioactivities. Although numerous literatures are available on the pharmacological and biological activities, the biological activity of SS in bone regeneration process has not yet been well-defined. Therefore, in this study, the effect of SS was investigated in the proliferation and differentiation of MC3T3-E1 osteoblastic cell line. The treatment of SS did not significantly affect the cell proliferation in MC3T3-E1 cells. SS significantly accelerated the mineralization and significantly increased the expression of alkaline phosphatase (ALP) and osteocalcin (OC) mRNAs, compared to the control, in the differentiation of MC3T3-E1 cells. SS significantly accelerated the decrease of osteonectin (ON) mRNA expression as compared with the control in a time-dependent manner in the differentiation of MC3T3-E1 cells. These results suggest that the SS facilitate the osteoblast differentiation and mineralization in MC3T3-E1 osteoblastic cells. Therefore, there may be potential properties for development and clinical application of bone regeneration materials.

Recently, techniques have been reported that involve the preparation of extracted teeth from patients used as particulated bone graft materials for bone graft purposes. For implant placement and bone graft, autogenous teeth bone graft materials were used in 15 patients, and clinically excellent results were obtained. In histological examination, favorable bony healing by osteoconduction was observed.

Highlights: • miR-663 is significantly up-regulated during MDPC-23 odontoblastic cell differentiation. • miR-663 accelerates mineralization in MDPC-23 odontoblastic cells without cell proliferation. • miR-663 promotes odontoblastic cell differentiation by targeting APC and activating Wnt/β-catenin signaling in MDPC-23 cells. - Abstract: MicroRNAs (miRNAs) regulate cell differentiation by inhibiting mRNA translation or by inducing its degradation. However, the role of miRNAs in odontogenic differentiation is largely unknown. In this present study, we observed that the expression of miR-663 increased significantly during differentiation of MDPC-23 cells to odontoblasts. Furthermore, up-regulation of miR-663 expression promoted odontogenic differentiation and accelerated mineralization without proliferation in MDPC-23 cells. In addition, target gene prediction for miR-663 revealed that the mRNA of the adenomatous polyposis coli (APC) gene, which is associated with the Wnt/β-catenin signaling pathway, has a miR-663 binding site in its 3′-untranslated region (3′UTR). Furthermore, APC expressional was suppressed significantly by miR-663, and this down-regulation of APC expression triggered activation of Wnt/β-catenin signaling through accumulation of β-catenin in the nucleus. Taken together, these findings suggest that miR-663 promotes differentiation of MDPC-23 cells to odontoblasts by targeting APC-mediated activation of Wnt/β-catenin signaling. Therefore, miR-663 can be considered a critical regulator of odontoblast differentiation and can be utilized formore » developing miRNA-based therapeutic agents.« less