Yong-Gun Kim

MicroRNA-124 regulates osteoclast differentiation.

Osteoclasts are specialized cells for bone-resorption originated from precursors of macrophage/monocyte lineage. The receptor activator of NFκB ligand (RANKL) initiates osteoclast differentiation, in which nuclear factor of activated T cell cytoplasmic 1 (NFATc1) plays a key role as a master transcription factor. In the present report, we show that microRNA-124 (miR-124) regulates osteoclastogenesis of mouse bone marrow macrophages (BMMs) by suppressing NFATc1 expression. On the other hand, synthetic inhibitor that binds specifically to miR-124 enhanced osteoclast differentiation and NFATc1 expression. The overexpression of a constitutively active form of NFATc1 prevented the inhibitory effect of miR-124 on osteoclastogenesis. Finally, miR-124 also affected the proliferation and motility of osteoclast precursors, the latter coinciding with the reduced expression of RhoA and Rac1. These findings not only reveal unprecedented role of miR-124 in osteoclastogenesis but also suggest a novel mode of regulation of NFATc1 in osteoclasts.

IL-17 inhibits osteoblast differentiation and bone regeneration in rat

OBJECTIVE The interleukin-17 (IL-17) family is a group of pro-inflammatory cytokines that are produced by a subset of helper T cells. IL-17 family members are not only involved in the immune response of tissues but also play a role in bone metabolism. Although the role of IL-17 in osteoclast-mediated bone resorption has been extensively studied, its role during osteoblast-mediated bone formation has rarely been investigated. In this study, we examined the effect of IL-17 on osteogenesis in rats both in vitro and in vivo. DESIGN To evaluate osteogenesis in vitro, rat calvarial osteoblast precursor cells were cultured for 14 days in osteogenic medium with or without 50ng/mL IL-17. Osteogenic activity was evaluated by alkaline phosphatase and alizarin red staining. The mRNA expression of alkaline phosphatase, osteocalcin, and osterix was also measured by using real-time PCR. To test whether IL-17 affects bone formation in vivo, bone filling was examined by micro-computed tomography and histological observations at 8 weeks after critical-sized defects were made in rat calvaria. RESULTS The presence of IL-17 significantly reduced alkaline phosphatase and alizarin red staining and the expression of alkaline phosphatase, osteocalcin, and osterix in vitro. IL-17 also significantly inhibited the filling of calvarial defects in vivo. CONCLUSION IL-17 exerted a negative effect on osteogenesis in a rat model. In contrast to the previously reported beneficial effect on osteogenic differentiation of human mesenchymal stem cells, our results suggest a species or cell type-specific role for IL-17 in bone formation.

Influence of crown-to-implant ratio on periimplant marginal bone loss in the posterior region: a five-year retrospective study.

Purpose The aim of this study was to evaluate the influence of the crown-to-implant (C/I) ratio on the change in marginal bone level around the implant and to determine the site-related factors influencing the relationship between the C/I ratio and periimplant marginal bone loss. Methods A total of 259 implants from 175 patients were evaluated at a mean follow-up of five years. Implants were divided into two groups according to their C/I ratios: ≤1, and >1. Site-related factors having an influence on the relationship between C/I ratio and periimplant marginal bone loss were analyzed according to the implant location, implant diameter, implant manufacturer, prosthesis type, and guided bone regeneration (GBR) procedure. Results It was found that 1) implants with a C/I ratio below 1 exhibited greater periimplant marginal bone loss than implants with a C/I ratio more than 1, 2) site-related factors had an effect on periimplant marginal bone loss, except for the implant system used, 3) the C/I ratio was the factor having more dominant influence on periimplant marginal bone loss, compared with implant diameter, prosthesis type, implant location, and GBR procedure, 4) implants with a C/I ratio below 1 showed greater periimplant marginal bone loss than implants with a C/I ratio greater than 1 in the maxilla, but not in the mandible, 5) and periimplant marginal bone loss was more affected by the implant system than the C/I ratio. Conclusions Within the limitations of this study, implants with a higher C/I ratio exhibited less marginal bone loss than implants with a lower C/I ratio in the posterior regions. The C/I ratio was a more dominant factor affecting periimplant marginal bone loss in the maxilla than the mandible. Meanwhile, the implant system was a more dominant factor influencing periimplant marginal bone loss than the C/I ratio.

The expression of a nitric oxide derivative, tissue inhibitors of metalloproteinase-3, and tissue inhibitors of metalloproteinase-4 in chronic periodontitis with type 2 diabetes mellitus

Purpose The purpose of this study was to analyze the expression of inducible nitric oxide synthases (iNOS), tissue inhibitors of metalloproteinase (TIMP)-3, and TIMP-4 in the gingival tissues of periodontal patients with or without type 2 diabetes mellitus (DM). Methods Depending on the patients systemic condition and clinical criteria of the gingiva, each gingival sample was classified into one of three groups. Sixteen clinically, systemically healthy patients (group 1), 16 periodontal patients (group 2), and 16 periodontal patients with DM (group 3) were included. Tissue samples in each group were collected, prepared, and analyzed by western blotting. Quantification of the relative amount of TIMP-3, TIMP-4, and iNOS was performed. Results The expression levels of iNOS and TIMP-3 both increased in group 1, group 2, and group 3 in increasing order, and were significantly higher in both group 2 and group 3 as compared to group 1 (P<0.05). The expression levels of TIMP-4 increased in the same order, but significantly increased in group 2 as compared to group 1, in group 3 as compared to group 1, and group 3 as compared to group 2 (P<0.05). Conclusions This study demonstrated that iNOS, TIMP-3, and TIMP-4 might be involved in the progression of periodontal inflammation associated with type 2 DM. It is thought that further study of these factors can be applied practically for the diagnosis and control of periodontitis in diabetics.

The effects of dexamethasone on the apoptosis and osteogenic differentiation of human periodontal ligament cells.

Purpose The purpose of the current study was to examine the effect of dexamethasone (Dex) at various concentrations on the apoptosis and mineralization of human periodontal ligament (hPDL) cells. Methods hPDL cells were obtained from the mid-third of premolars extracted for orthodontic reasons, and a primary culture of hPDL cells was prepared using an explant technique. Groups of cells were divided according to the concentration of Dex (0, 1, 10, 100, and 1,000 nM). A 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay was performed for evaluation of cellular viability, and alkaline phosphatase activity was examined for osteogenic differentiation of hPDL cells. Alizarin Red S staining was performed for observation of mineralization, and real-time polymerase chain reaction was performed for the evaluation of related genes. Results Increasing the Dex concentration was found to reduce cellular viability, with an increase in alkaline phosphatase activity and mineralization. Within the range of Dex concentrations tested in this study, 100 nM of Dex was found to promote the most vigorous differentiation and mineralization of hPDL cells. Dex-induced osteogenic differentiation and mineralization was accompanied by an increase in the level of osteogenic and apoptosis-related genes and a reduction in the level of antiapoptotic genes. The decrease in hPDL cellular viability by glucocorticoid may be explained in part by the increased prevalence of cell apoptosis, as demonstrated by BAX expression and decreased expression of the antiapoptotic gene, Bcl-2. Conclusions An increase in hPDL cell differentiation rather than cellular viability at an early stage is likely to be a key factor in glucocorticoid induced mineralization. In addition, apoptosis might play an important role in Dex-induced tissue regeneration; however, further study is needed for investigation of the precise mechanism.

Bone dynamics in the upward direction after a maxillary sinus floor elevation procedure: serial segmentation using synchrotron radiation micro-computed tomography

Objective Maxillary sinus floor augmentation has been shown to be the most predictable surgical technique for enhancing the bone volume in the posterior area of the maxilla. The purpose of this study was to analyze the serial slice image segmentation of newly formed bone and bone substitutes after sinus floor elevation using synchrotron radiation X-ray micro-computed tomography (SR-μCT). Materials and methods Bone biopsy specimens were collected after 6 months of sinus floor augmentation. From the six bone biopsy specimens, the cross-sectional images at every 8 μm along the apical direction from the inferior border using serial segmentation from three-dimensional reconstructed X-ray images were analyzed. The amount of new bone and bone substitutes were measured at each slicing image (300–430 images per specimen). Results The bone dynamics between the new bone and bone substitutes along the inferior–superior direction in humans after maxillary sinus floor elevation (MSFE) were analyzed using the whole sample region. Although these observations suggest that the specimens are structurally inhomogeneous, sinus floor elevation was confirmed to be a reliable surgical procedure for increasing the amount of bone. Conclusion SR-μCT is highly effective for obtaining high-resolution images. An analysis of biological specimens using SR-μCT is quite reliable and this technique will be an important tool in the wide field of tissue engineering.

Fluvastatin Inhibits Osteoclast Differentiation and Porphyromonas gingivalis Lipopolysaccharide-Induced Alveolar Bone Erosion in Mice

BACKGROUND Statins have been widely used to treat hypercholesterolemia. In addition to inhibition of cholesterol synthesis, recent reports suggest a bone anabolic property of statins. However, little notice has been paid to the direct effect of statins on osteoclastogenesis and bone resorption. METHODS The effect of fluvastatin on osteoclast differentiation was determined using in vitro culture of mouse bone marrow macrophages (BMMs) in the presence of macrophage colony-stimulating factor and receptor activator of nuclear factor-kappa B ligand (RANKL). The role of fluvastatin on bone erosion was examined in the Porphyromonas gingivalis (Pg) lipopolysaccharide (LPS)-induced alveolar bone loss model in mice. RESULTS Fluvastatin significantly inhibited both RANKL- and LPS-induced osteoclast differentiation in mouse BMMs. Fluvastatin also markedly reduced expression of osteoclast differentiation marker genes Acp5, Calcr, and Ctsk as well as fusion markers Atp6v0d2 and Dcstamp. These were accompanied by decreased expression of c-Fos and nuclear factor of activated T cells cytoplasmic 1 transcription factors. Fluvastatin reduced generation of reactive oxygen species upon the addition of RANKL and LPS, suggesting an antioxidant role. Finally, administration of fluvastatin in mice conspicuously reduced Pg LPS-induced osteoclastogenesis and alveolar bone erosion in vivo. CONCLUSION Combined, these results suggest fluvastatin directly inhibited osteoclastogenesis and efficiently blocked bone erosion.

Effects of Escherichia Coli-derived Recombinant Human Bone Morphogenetic Protein-2 Loaded Porous Hydroxyaptite-based Ceramics on Calvarial Defect in Rabbits

Background Recombinant human bone morphogenetic proteins (rhBMPs) have been widely used in regenerative therapies to promote bone formation. The production of rhBMPs using bacterial systems such as Escherichia coli (E. coli) is estimated to facilitate clinical applications by lowering the cost without compromising biological activity. In clinical practice, rhBMP-2 and osteoconductive carriers (e.g., hydroxyapatite [HA] and bovine bone xenograft) are used together. This study examined the effect of E. coli-derived rhBMP-2 combined with porous HA-based ceramics on calvarial defect in rabbits. Methods Six adult male New Zealand white rabbits were used in this study. The experimental groups were divided into the following 4 groups: untreated (NC), bovine bone graft (BO), porous HA (HA) and porous HA with rhBMP-2 (HA-BMP). Four transosseous defects of 8 mm in diameter were prepared using stainless steel trephine bur in the frontal and parietal bones. Histological and histomorphometric analyses at 4 weeks after surgery revealed significant new bone formation by porous HA alone. Results HA-BMP showed significantly higher degree of bone formation compared with BO and HA group (P<0.05). The average new bone formation % (new bone area per total defect area) of NC, BO, HA, and HA-BMP at 4-week after surgery were 12.65±5.89%, 29.63±6.99%, 28.86±6.17% and 49.56±8.23%, respectively. However, there was no statistical difference in the bone formation between HA and BO groups. Conclusions HA-BMP promoted more bone formation than NC, BO and HA alone. Thus, using E. coli-derived rhBMP-2 combined with porous HA-based ceramics can promote new bone formation.

Long-term evaluation of teeth and implants during the periodic maintenance in patients with viral liver disease

PURPOSE This study was designed to investigate the maintenance of teeth and implants in patients with viral liver disease. MATERIALS AND METHODS 316 patients without any significant systemic disease were selected as a control group. Liver disease group was consisted of 230 patients. Necessary data were collected using clinical records and panoramic radiographs. Then, the patients were subdivided into 2 groups based on the type of active dental therapy received before maintenance period (Pre-Tx). Analysis for finding statistically significant difference was performed based on the need for re-treatment of active dental therapy (Re-Tx) and change in the number of teeth (N-teeth) and implants (N-implants). RESULTS Comparing to control group, the patients with liver disease showed higher value on N-teeth, N-implants, and Re-Tx. Statistically significant differences were found on N-teeth (P=.000) and Re-Tx (P=.000) in patients with non-surgical Pre-Tx. Analysis based on severity of liver disease showed that N-teeth and Re-Tx were directly related to severity of liver disease regardless of received type of Pre-Tx. Significant differences were found on N-teeth (P=.003) and Re-Tx (P=.044) in patients with non-surgical Pre-Tx. CONCLUSION In this study, it was concluded that liver disease might influence the loss of teeth and cause the relapse of dental disease during maintenance period in patients. A significant positive relationship between tooth and implant loss and severity of liver disease seems to exist.

Transcriptome sequencing of gingival biopsies from chronic periodontitis patients reveals novel gene expression and splicing patterns

BackgroundPeriodontitis is the most common chronic inflammatory disease caused by complex interaction between the microbial biofilm and host immune responses. In the present study, high-throughput RNA sequencing was utilized to systemically and precisely identify gene expression profiles and alternative splicing.MethodsThe pooled RNAs of 10 gingival tissues from both healthy and periodontitis patients were analyzed by deep sequencing followed by computational annotation and quantification of mRNA structures.ResultsThe differential expression analysis designated 400 up-regulated genes in periodontitis tissues especially in the pathways of defense/immunity protein, receptor, protease, and signaling molecules. The top 10 most up-regulated genes were CSF3, MAFA, CR2, GLDC, SAA1, LBP, MME, MMP3, MME-AS1, and SAA4. The 62 down-regulated genes in periodontitis were mainly cytoskeletal and structural proteins. The top 10 most down-regulated genes were SERPINA12, MT4, H19, KRT2, DSC1, PSORS1C2, KRT27, LCE3C, AQ5, and LCE6A. The differential alternative splicing analysis revealed unique transcription variants in periodontitis tissues. The EDB exon was predominantly included in FN1, while exon 2 was mostly skipped in BCL2A1.ConclusionsThese findings using RNA sequencing provide novel insights into the pathogenesis mechanism of periodontitis in terms of gene expression and alternative splicing.