25-Hydroxycholesterol induces odontoclastic differentiation through RANK-RANKL upregulation and NF-κB activation in odontoblast-like MDPC-23 cells: An in vitro study.

AIM: The physiological effects and cellular mechanism of 25-hydroxycholesterol (25-HC), which is an oxysterol synthesized from cholesterol by cholesterol-25-hydroxylase (CH25H) expressed under inflammatory conditions, are still largely unknown during odontoclastogenesis. This study aimed to evaluate 25-HC-induced odontoclastogenesis and its cellular mechanisms in odontoblast-like MDPC-23 cells. METHODOLOGY: To investigate 25-HC-induced odontoclastogenesis of MDPC-23 cells and its cellular mechanism, haemotoxylin and eosin staining, tartrate-resistant acid phosphatase (TRAP) staining, dentine resorption assay, zymography, reactive oxygen species (ROS) detection, immunocytochemistry, and nuclear translocation were performed. The experimental values are presented as mean ± standard deviation and were compared using analysis of variance, followed by post hoc multiple comparisons (Tukey's test) using SPSS software version 22 (IBM Corp.). A p-value <.05 was considered statistically significant. RESULTS: Lipopolysaccharide or receptor activator of nuclear factor-κB ligand (RANKL) induced the synthesis of 25-HC via the expression of CH25H in MDPC-23 cells (p < .01). Multinucleated giant cells with morphological characteristics and TRAP activity of the odontoclast were increased by 25-HC in MDPC-23 cells (p < .01). Moreover, 25-HC increased dentine resorption through the expression and activity of matrix metalloproteinases in MDPC-23 cells. It not only increased the expression of odontoclastogenic biomarkers but also translocated cytosolic nuclear factor-κB (NF-κB) to the nucleus in MDPC-23 cells. Additionally, 25-HC not only increased the production of ROS (p < .01), expression of inflammatory mediators (p < .01), pro-inflammatory cytokines, receptor activator of NF-κB (RANK), and RANKL but also suppressed the expression of osteoprotegerin (OPG) in MDPC-23 cells. In contrast, CDDO-Me, a chemical NF-κB inhibitor, decreased TRAP activity (p < .01) and downregulated the expression of the odontoclastogenic biomarkers, including RANK and RANKL, in MDPC-23 cells. CONCLUSION: 25-HC induced odontoclastogenesis by modulating the RANK-RANKL-OPG axis via NF-κB activation in MDPC-23 cells. Therefore, these findings provide that 25-HC derived from cholesterol metabolism may be involved in the pathophysiological etiological factors of internal tooth resorption.

Volumetric evaluation of effects of platelet-rich fibrin and concentrated growth factor on early bone healing after endodontic microsurgery: a randomized controlled trial.

BACKGROUND: This randomized controlled clinical trial compared the effects of platelet-rich fibrin (PRF) and concentrated growth factor (CGF) on early bone healing after endodontic microsurgery. METHODS: Eighteen patients with an isolated periapical lesion < 10 mm in the maxillary anterior region were randomly assigned to three groups: control, PRF, or CGF. Endodontic microsurgery was performed and PRF or CGF membranes were placed over the bone defects in the experimental groups. The volume of the bone defect at postoperative one week, three months, and six months was evaluated using cone-beam computed tomography and Mimics software. The results were statistically analyzed using the Kruskal-Wallis test and post-hoc Mann-Whitney U test with Bonferroni correction. RESULTS: At the three-month follow-up, the PRF and CGF groups showed significantly greater bone healing compared with the control group (p > 0.05). However, no significant difference was observed between the PRF and CGF groups. At the six-month follow-up, no significant differences were observed between the groups. CONCLUSIONS: These results suggested that PRF and CGF promote early bone healing after endodontic microsurgery.

Technical Note on Simplified Free Gingival Graft Using Tack Fixation (sFGG).

Free gingival graft (FGG) is the gold standard procedure for the reliable augmentation of lost keratinized mucosa (KM) around dental implants. This conventional surgical approach has its drawbacks, including limitations in manipulation, the requirement for suturing, postoperative discomfort, and pain. This case report aimed to evaluate the efficacy of a simplified free gingival graft (sFGG) in addressing the issue of inadequate keratinized mucosa around dental implants. Fixation tacks were used to perform the sFGG procedure. Initially, a partial-thickness flap was created and apically repositioned. The gingival graft was harvested from the palate with a narrow profile and securely affixed to the recipient site using 5 mm long fixation tacks. Significant gains in keratinized mucosa were achieved and successfully maintained within 1 year. Consequently, the sFGG technique emerges as a simple and reliable treatment approach for managing inadequate keratinized mucosa around dental implants.

GPR183 Regulates 7α,25-Dihydroxycholesterol-Induced Oxiapoptophagy in L929 Mouse Fibroblast Cell.

7α,25-dihydroxycholesterol (7α,25-DHC) is an oxysterol synthesized from 25-hydroxycholesterol by cytochrome P450 family 7 subfamily B member 1 (CYP7B1) and is a monooxygenase (oxysterol-7α-hydroxylase) expressed under inflammatory conditions in various cell types. In this study, we verified that 7α,25-DHC-induced oxiapoptophagy is mediated by apoptosis, oxidative stress, and autophagy in L929 mouse fibroblasts. MTT assays and live/dead cell staining revealed that cytotoxicity was increased by 7α,25-DHC in L929 cells. Consequentially, cells with condensed chromatin and altered morphology were enhanced in L929 cells incubated with 7α,25-DHC for 48 h. Furthermore, apoptotic population was increased by 7α,25-DHC exposure through the cascade activation of caspase-9, caspase-3, and poly (ADP-ribose) polymerase in the intrinsic pathway of apoptosis in these cells. 7α,25-DHC upregulated reactive oxygen species (ROS) in L929 cells. Expression of autophagy biomarkers, including beclin-1 and LC3, was significantly increased by 7α,25-DHC treatment in L929 cells. 7α,25-DHC inhibits the phosphorylation of Akt associated with autophagy and increases p53 expression in L929 cells. In addition, inhibition of G-protein-coupled receptor 183 (GPR183), a receptor of 7α,25-DHC, using GPR183 specific antagonist NIBR189 suppressed 7α,25-DHC-induced apoptosis, ROS production, and autophagy in L929 cells. Collectively, GPR183 regulates 7α,25-DHC-induced oxiapoptophagy in L929 cells.

Vertical bony step between proximal and distal segments after mandibular setback is related with relapse: A cone-beam computed tomographic study.

INTRODUCTION: Vertical bony step (VBS) occurs between proximal and distal segments of the mandible during mandibular setback surgery with bilateral sagittal split ramus osteotomy. The purpose of this study was to investigate whether VBS is correlated with the relapse of mandibular setback using 3-dimensional models constructed from cone-beam computed tomography. METHODS: The subjects consisted of 30 patients who underwent bilateral sagittal split ramus osteotomy for a mandibular setback. Double jaw surgery was performed in 18 patients, and isolated mandibular setback surgery was performed in 12 patients. Cone-beam computed tomography scans were taken at pretreatment (T0), postsurgery (T1), and posttreatment (T2). Treatment changes and the correlations between measurements were evaluated. RESULTS: The mean mandibular setback was -11.9 mm, and the mean VBS was -5.6 mm. Correlations with the relapse of mandibular setback were found in the amount of mandibular setback (T1 - T0), development of VBS (T1 - T0), posterior movement of the proximal segment (T1 - T0), counterclockwise rotation of symphysis (T2 - T1), and the resolution of VBS (T2 - T1). CONCLUSIONS: The development and resolution of VBS were correlated with the relapse of mandibular setback. Minimizing VBS is recommended to reduce the relapse of mandibular setback.

25-Hydroxycholesterol-Induced Oxiapoptophagy in L929 Mouse Fibroblast Cell Line.

25-hydroxycholesterol (25-HC) is an oxysterol synthesized from cholesterol by cholesterol-25-hydroxylase during cholesterol metabolism. The aim of this study was to verify whether 25-HC induces oxiapoptophagy in fibroblasts. 25-HC not only decreased the survival of L929 cells, but also increased the number of cells with condensed chromatin and altered morphology. Fluorescence-activated cell sorting results showed that there was a dose-dependent increase in the apoptotic populations of L929 cells upon treatment with 25-HC. 25-HC-induced apoptotic cell death was mediated by the death receptor-dependent extrinsic and mitochondria-dependent intrinsic apoptosis pathway, through the cascade activation of caspases including caspase-8, -9, and -3 in L929 cells. There was an increase in the levels of reactive oxygen species and inflammatory mediators such as inducible nitric oxide synthase, cyclooxygenase-2, nitric oxide, and prostaglandin E2 in L929 cells treated with 25-HC. Moreover, 25-HC caused an increase in the expression of beclin-1 and microtubule-associated protein 1A/1B-light chain 3, an autophagy biomarker, in L929 cells. There was a significant decrease in the phosphorylation of protein kinase B (Akt) in L929 cells treated with 25-HC. Taken together, 25-HC induced oxiapoptophagy through the modulation of Akt and p53 cellular signaling pathways in L929 cells.

Oxysterol 25-hydroxycholesterol as a metabolic pathophysiological factors of osteoarthritis induces apoptosis in primary rat chondrocytes.

The aim of the present study was to investigate the pathophysiological etiology of osteoarthritis that is mediated by the apoptosis of chondrocytes exposed to 25-hydroxycholesterol (25-HC), an oxysterol synthesized by the expression of cholesterol-25-hydroxylase (CH25H) under inflammatory conditions. Interleukin-1ß induced the apoptosis of chondrocytes in a dose- dependent manner. Furthermore, the production of 25-HC increased in the chondrocytes treated with interleukin-1ß through the expression of CH25H. 25-HC decreased the viability of chondrocytes. Chondrocytes with condensed nucleus and apoptotic populations increased by 25-HC. Moreover, the activity and expression of caspase-3 were increased by the death ligand-mediated extrinsic and mitochondria-dependent intrinsic apoptotic pathways in the chondrocytes treated with 25-HC. Finally, 25-HC induced not only caspase-dependent apoptosis, but also induced proteoglycan loss in articular cartilage ex vivo cultured rat knee joints. These data indicate that 25-HC may act as a metabolic pathophysiological factor in osteoarthritis that is mediated by progressive chondrocyte death in the articular cartilage with inflammatory condition.

Anticatabolic Effects of Morin through the Counteraction of Interleukin-1ß-Induced Inflammation in Rat Primary Chondrocytes.

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-1ß (IL-1ß) 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-1ß-induced decrease in extracellular matrix. Furthermore, treatment with morin attenuated IL-1ß-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.

Effects of Platelet-Derived Material (Platelet-Rich Fibrin) on Bone Regeneration.

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.

Bilateral supernumerary clavicular heads of sternocleidomastoid muscle in a Korean female cadaver.

Many anatomical variants on the sternocleidomastoid muscle have been reported. In this study, supernumerary clavicular heads of sternocleidomastoid muscle in a Korean female cadaver were bilaterally displayed. The observed supernumerary heads were classified as follows: one sterno-mastoid, one cleido-occipital and one cleido-mastoid on the right side, and one sterno-mastoid-occipital, four cleido-occipitals, and one cleido-mastoid on the left side. The sterno-mastoid and sterno-mastoid-occipital and the cleido-occipital made the superficial layer of the sternocleidomastoid muscle, while others made deep layer. We discussed clinical relevance and developmental basis of these muscular variations important for clinicians and anatomists.

Comparative Study on Early Osseointegration of Implants According to Various Drilling Speeds in the Mandible of Dogs.

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.

Comparative Study on Osseointegration of Implants After Flap and Flapless Surgery in the Mandible of Dogs.

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.

Incidence and Management of Fractured Dental Implants: Case Reports.

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.

Sinus Membrane Elevation by the Crestal Approach Using a Novel Drilling System.

PURPOSE: 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. MATERIALS AND METHODS: 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). RESULTS: 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). CONCLUSION: 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.

Synthesis and Characterization of ß-Tricalcium Phosphate Derived From Haliotis sp. Shells.

PURPOSE: To develop a methodology for the synthesis of ß-tricalcium phosphate (ß-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, ß-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 ß-TCP was successfully synthesized from abalone shell. The synthesized ß-TCP did not affect cell viability of either normal human oral keratinocytes or osteoblastic MG-63 cells. These data indicate that ß-TCP synthesized from abalone shell is biologically safe. CONCLUSIONS: ß-TCP (Ca3(PO4)2) synthesized from abalone shell can be used as a potential source of bone grafting material.

Biochanin-A antagonizes the interleukin-1ß-induced catabolic inflammation through the modulation of NFκB cellular signaling in primary rat chondrocytes.

Biochanin-A, a phytoestrogen derived from herbal plants, protected from the IL-1ß-induced loss of proteoglycans through the suppression of matrix degrading enzymes such as matrix metalloproteinase (MMP)-13, MMP-3, MMP-1, and ADAMTS-5 in primary rat chondrocytes and the knee articular cartilage. It also suppressed the expression of IL-1ß-induced catabolic factors such as nitric oxide synthase 2, cyclooxygenase-2, prostaglandin E2, and inflammatory cytokines. Furthermore, biochanin-A suppressed the IL-1ß-induced phosphorylation of NFκB, and inhibited its nuclear translocation in primary rat chondrocytes. These results indicate that biochanin-A antagonizes the IL-1ß-induced catabolic effects through its anti-inflammatory activity that involves the modulation of NFκB signaling.

Implants Displaced Into the Maxillary Sinus: A Systematic Review.

OBJECTIVES: 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. MATERIALS AND METHODS: 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. RESULTS: 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. CONCLUSION: 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.

Histomorphometric Analysis of Contaminated Autogenous Tooth Graft Materials After Various Sterilization.

PURPOSE: The purpose of this study was to evaluate histomorphometrically contaminated autogenous tooth graft materials, which were resterilized. MATERIALS AND METHODS: The intentional defects (diameter: 8 mm, depth: 4 mm) were formed around implant fixture on the iliac crest of 6 mongrel dogs. Autogenous tooth graft materials were made by extracted premolars. After the contamination of the tooth materials, graft procedure was performed; no contaminated group (control group), contaminated groups (nonsterilization group [group 1], ethylene oxide [EO] gas group [group 2], and autoclave group [group 3]). The bone-to-implant contact (BIC) and the new bone formation rate (NBFR) were evaluated after sacrifice. RESULTS: The BIC and NBFR of groups 1 and 3 were significantly lower than the control group after 4 weeks. The BIC and NBRF of group 3 were significantly lower than the control group after 8 weeks. However, the BIC and NBRF of group 2 was not significantly different comparing with the control group after 4 and 8 weeks. CONCLUSION: Sterilization using EO gas may be more favorable than high-pressure sterilization in cases the reuse of contaminated autogenous tooth graft materials.

Early Bone Formation at a Femur Defect Using CGF and PRF Grafts in Adult Dogs: A Comparative Study.

PURPOSE: The purpose of this study was to compare the predictability of new bone formation using an autologous concentrated growth factor (CGF) graft alone and platelet graft alone. MATERIALS AND METHODS: Four bony defects of 8 mm were formed, and 3.7- × 10-mm implants were placed in the right femur. The platelet-rich fibrin (PRF), CGF, and synthetic bone were grafted to the bone defect area. Enzyme linked immunosorbent assay quantitative analysis and microscopic analysis of the fibrinogen structure were performed. RESULTS: At 4 weeks, the comparisons of each experimental group showed a significant difference between the CGF group and the synthetic bone graft group. When comparing the CGF and allograft material groups, the allograft group showed significantly more new bone formation. In the case of vascular endothelial growth factor, CGF had 1.5 times more than PRF. CGF showed a fibrinogen structure with a constant diameter. CONCLUSION: When applied to a clinical case, CGF is predicted to show better results than PRF.

Biological Effects of the Herbal Plant-Derived Phytoestrogen Bavachin in Primary Rat Chondrocytes.

The aim of this study was to examine the anabolic and anticatabolic functions of bavachin in primary rat chondrocytes. With bavachin treatment, chondrocytes survived for 21 d without cell proliferation, and the proteoglycan content and extracellular matrix increased. Short-term monolayer culture of chondrocytes showed that gene induction of both aggrecan and collagen type II, major extracellular matrix components, was significantly upregulated by bavachin. The expression and activities of cartilage-degrading enzymes such as matrix metalloproteinases and a disintegrin and metalloproteinase with thrombospondin motifs were inhibited significantly by bavachin, while tissue inhibitors of metalloprotease were significantly upregulated. Bavachin inhibits the expression of inducible nitric oxide synthase, a representative catabolic factor, and downregulated the expression of nitric oxide, cyclooxygenase-2, and prostaglandin E2 in a dose-dependent manner in chondrocytes. Our results suggest that the bavachin has anabolic and potent anticatabolic biological effects on chondrocytes, which may have considerable promise in treating articular cartilage degeneration in the future.