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.

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.

A Retrospective Analysis of the Retreatment of Failed Sinus Bone Grafts.

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.

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.

Development of an Experimental Animal Model for Lower Back Pain by Percutaneous Injury-Induced Lumbar Facet Joint Osteoarthritis.

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.

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.

A Comparative Study of Stability After the Installation of 2 Different Surface Types of Implants in the Maxillae of Dogs.

PURPOSE: This study was performed to investigate the histologic and histomorphometric findings of 2 different types of implant. MATERIALS AND METHODS: Resorbable blasting media (RBM) and sandblasted with larger grit and acid etched (SLA) surfaced implants (24 fixtures in each group) were installed in posterior maxilla of dogs. The initial stability was measured using Periotest (Periotest value [PTV]). After 6 or 12 weeks, fixtures with surrounding bone were harvested. RESULTS: The average initial stability of the SLA group (-1.71 ± 2.9) was higher than that of the RBM group (-1.25 ± 3.21), but there was no significant difference. The mean PTV of the RBM surface was higher than the SLA surface at 12 weeks. The average bone-implant contacts were 67.6% ± 16.0% at 6 weeks and 82.7% ± 8.6% at 12 weeks in the SLA group and 69.9% ± 17.6% at 6 weeks and 78.3% ± 9.2% at 12 weeks in the RBM group. CONCLUSION: The SLA and resorbable blasting media (RBM) surface implants demonstrated good stabilities and healing processes of the surrounding bone in the posterior maxilla. Therefore, the two domestic implants could provide predictable clinical results.

Implant Stability Measurements in the Long-Term Follow-up of Dentis Implants: A Retrospective Study With Periotest.

PURPOSE: The purpose of this study was to evaluate retrospectively the stability of Dentis implant with the Periotest. METHODS: In total, 36 patients and 88 implants were investigated. Periotest was used to measure implant stability, and a periapical view was taken immediately after surgery and again immediately after, 3 months after, 6 months and 5 years after prosthesis placement. Bone loss on the periapical view, bone quality according to tactile sensation, and area of implant installation were assessed. RESULTS: The mean Periotest value (PTV) immediately after surgery was -1.02, and the mean bone loss rate (bone loss/fixture length × 100) at 6 months after prosthesis placement was 8.42%. PTV was higher with more bone loss (types III, IV vs types I, II bone). The lowest mean PTV was in the lower molar area (-1.48), followed by the lower anterior (-1.41), upper molar (0.11), and upper anterior area (5). One implant failed and survival rates were 98.9%. CONCLUSION: Implant stability was lower in cases with more bone loss and poor bone quality and in the maxilla versus the mandible.

Evaluation of bone formation after grafting with deproteinized bovine bone and mineralized allogenic bone.

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.

Downregulation of adenomatous polyposis coli by microRNA-663 promotes odontogenic differentiation through activation of Wnt/beta-catenin signaling.

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/ß-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/ß-catenin signaling through accumulation of ß-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/ß-catenin signaling. Therefore, miR-663 can be considered a critical regulator of odontoblast differentiation and can be utilized for developing miRNA-based therapeutic agents.

MicroRNA-205 suppresses the oral carcinoma oncogenic activity via down-regulation of Axin-2 in KB human oral cancer cell.

MicroRNA (miRNA) is a small noncoding RNA molecule, 19-25 nucleotides in length, which regulates several pathways including cell development, cell proliferation, carcinogenesis, apoptosis, etc. In this study, the over-expression of microRNA-205 (miR-205) increased the number of apoptotic cells by at least 4 times compared to the control. In addition, over-expressed miRNA in KB oral cancer cells triggered apoptosis via the caspase cascade, including the cleavage of caspase-9, caspase-7, caspase-3, and PARP. Flow cytometry showed that apoptotic cell death was increased significantly by 35.33% in KB oral cancer cells with over-expressed miR-205 compared to the control. The microarray data showed that axis inhibitor protein 2 (Axin2) was down-regulated in KB oral cancer cells transfected with miR-205. In addition, Axin2 was down-regulated by approximately 50% by over-expressed miR-205 at both the mRNA and protein levels. Interestingly, Axin2 was up-regulated in KB oral cancer compared to human normal oral keratinocytes. Furthermore, the cell cytotoxicity and apoptotic population of KB oral cancer cells were increased significantly after Axin2 siRNA transfection. These results suggest that Axin2 is might be as potential oncogene in KB oral cancer cells. The luciferase assay showed that over-expressed miR-205 in KB oral cancer cells suppressed AXIN2 expression through an interaction with its own binding site at AXIN2 3'UTR (64-92). These results suggest that miR-205 is a novel anti-oncogenic miRNA in KB oral cancer cells, and may have potential applications in oral cancer therapy.

JPH203, an L-type amino acid transporter 1-selective compound, induces apoptosis of YD-38 human oral cancer cells.

Compared to most normal cells that express L-type amino acid transporter 2, L-type amino acid transporter 1 is highly expressed in cancer cells and presumed to support their elevated growth and proliferation. This study examined JPH203, a potent and selective L-type amino acid transporter 1 inhibitor, and its ability to suppress YD-38 human oral cancer cell growth. The YD-38 cells express L-type amino acid transporter 1 with its associating protein 4F2 heavy chain, but not L-type amino acid transporter 2. JPH203 and BCH, a non-selective L-type amino acid transporter inhibitor, completely inhibited l-leucine uptake in YD-38 cells. As expected, the intrinsic affinity of JPH203 to inhibit l-leucine uptake was far more efficient than BCH. Likewise, JPH203 and BCH inhibited YD-38 cell growth, with JPH203 being superior to BCH. JPH203 up-regulated the population of apoptotic YD-38 cells through the activation of apoptotic factors, including caspases and PARP. These results suggest that the inhibition of L-type amino acid transporter 1 activity via JPH203, which may act as a potential novel anti-oral-cancer agent, leads to apoptosis by inducing the intracellular depletion of the neutral amino acids essential for cancer cell growth in YD-38 human oral cancer cells.