Objectives : The aim of this study was to evaluate the efficacy of Aconiti Lateralis Preparata Radix (AP) and Acanthopanacis Cortex (AT) extracts in bone-derived adipocyte OP9 cell, osteoclast and osteoblast-like MG63 cells. Methods : MTT assay was used to evaluate the cytotoxicity of AP and AT extracts on OP9, osteoclast and MG63 cells. OP9 cells were treated with AP and AT, and the alterations in fat storage in the cells were determined by the Oil red O. To explain effects of RANKL-induced osteoclast differentiation in bone marrow macrophages, we performed the TRAP staining. The protein level of CAAAT/enhancer binding protein alpha ($C/EBP{\alpha}$) and peroxisome proliferator-activated receptor ${\gamma}$ ($PPAR{\gamma}$) as a adipocyte differentiation marker, and adiponectin was examined using western blot in differentiated OP9 cells. Effects of related genes were confirmed by luciferase assay using reporter assay. Results : AP and AT was not toxic on OP9 and MG63 cells, but AT was a little cytotoxic to osteoclast at the dose of $100{\mu}g/m{\ell}$. They could inhibit differentiation of OP9 cells and osteoclast with results of oil red O staining and TRAP staining. By western blot, AP and AT decreased the expression of $PPAR{\gamma}$ and $C/EBP{\alpha}$ which is the key transcription factor in adipogenesis and adiponectin secretion. AT also inhibited the BMP-4 activity in luciferase assay. AP also inhibited BMP-4 and Wnt3a activity, stimulated ER-${\beta}$ activity but inhibited androgen receptor activity. Conclusions : These results show AP and AT can be useful in osteoporosis and obesity via inhibition of osteoclast and adipocyte differentiation.
Kim, Yang-Hee;Yang, Hun-Mo;Song, Ho-Yeon;Lee, Byong-Taek
Proceedings of the Materials Research Society of Korea Conference
/
2010.05a
/
pp.44.1-44.1
/
2010
poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) is a bacterially derived copolymer produced by fermentation. PHBV has been attractive because of its potential environmental, pharmaceutical and biomedical applications. Recently, the electrospinning technique has been used to fabricate fibrous mat for biomedical applications such as artificial blood vessel, drug release and scaffolds, because this method is simple and easy to get ultrafine polymer fibers. Depending on speed of rotation drum collector, fiber structure was different. In this work, PHBV fiber was aligned by electrospinnning machine. Furthermore, alignment of PHBV fiber mats was given angle such as $45^{\circ}$, $60^{\circ}$ and $90^{\circ}$. The morphology of each aligned PHBV fiber mat was observed by SEM technique. The mechanical property was evaluated depending on alignment angle. Especially, cell attachment ability depending on alignment of PHBV fiber mats was carried out using MG- 63 osteoblast like cells.
Proceedings of the Korean Institute of Surface Engineering Conference
/
2018.06a
/
pp.24-24
/
2018
One of the challenges in tissue engineering is the design of optimal biomedical scaffolds, which can be governed by topographical surface characteristics, such as size, shape, and direction. Of these properties, we focus on the effects of nano - to micro - sized hierarchical surface. To fabricate the hierarchical surface structure on poly(${\varepsilon}$-caprolactone) (PCL) film, we employed a nano/micro-casting technique (NCT) and modified plasma process. The micro size topography of PCL film was controlled by sizes of the micro structures on lotus leaf. Also, the nano-size topography and hydrophilicity of PCL film were controlled by modified plasma process. After the plasma treatment, the hydrophobic property of the PCL film was significantly changed into hydrophilic property, and the nano-sized structure was well developed, as increasing the plasma exposure time and applied power. The surface properties of the modified PCL film were investigated in terms of initial cell morphology, attachment, and proliferation using osteoblast-like-cells (MG63). In particular, initial cell attachment, proliferation and osteogenic differentiation in the hierarchical structure were enhanced dramatically compared to those of the smooth surface.
Journal of the Korean Society of Manufacturing Process Engineers
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v.21
no.4
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pp.70-76
/
2022
Calcium-phosphate-based bioceramics are promising biomaterials for scaffolds because they can assist in bone regeneration. In this study, a laser sintering deposition system was developed, and 3D hydroxyapatite (HA) scaffolds were fabricated. The main process conditions of the HA scaffolds were laser power, table velocity, and laser focal distance. As the laser power increased, the line width, line height, and layer thickness also increased. Further, the line width, line height, and layer thickness decreased as the table velocity increased. As the laser focal distance increased, the line width increased, but the line height and layer thickness decreased. The fabricated green scaffolds were sintered at 1050 ℃ and 1150 ℃. The sintered scaffolds had a uniform and continuous interconnected shape, with pore sizes ranging from 850 to 950 ㎛ having 53% porosity. The compressive strength of the scaffolds decreased from 0.72 MPa (1050 ℃) to 0.53 MPa (1150 ℃). The biocompatibility of the scaffolds was investigated by analyzing the adhesion of osteoblast-like MG-63 cells cultured on the surfaces of the scaffolds. The results indicate that the scaffold sintered at 1050 ℃ had good mechanical and biological properties compared to that at 1150 ℃.
Kim, Il-Yeon;Jung, Sung-Min;Hwang, Soon-Jung;Shin, Sang-Wan
The Journal of Korean Academy of Prosthodontics
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v.47
no.4
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pp.376-384
/
2009
Purpose: The purpose of this study was to evaluate the response of osteoblast-like cells to Ca-P coated surface obtained via Ion beam-assisted deposition (IBAD) method and Sol-Gel process on anodized surface by cellular proliferation and differentiation. Material and methods: The surface of a commercially pure titanium (Grade IV) discs with dimension of 10mm diameter and 2 mm thickness was modified by anodic oxidation under a constant voltage of 300 V. The experimental groups were coated with Ca-P by the IBAD method and Sol-Gel process on anodized surface. The surface roughness (Ra) of specimens was measured by optical interferometer and each surface was examined by SEM. To evaluate cell response, MG63 cells were cultured and cell proliferation, ALP activity and the ability of cell differentiation were examined. Also, cell morphology was examined by SEM. The significant of each group was verified by Kruskal-Wallis Test ($\alpha$=.05). Results: The Ra value of Ca-P coated surface by IBAD method was significantly higher than Ca-P coated surface by Sol-gel process (P < .05). The level of cell proliferation and ALP activity was higher in Ca-P coated surface by IBAD method (P<.05). The expression of ALP showed higher level expression in Ca-P coated surface by IBAD method. Cells grown on Ca-P coated surface by IBAD method were uniformly distributed and developed a very close layer. Conclusion: These experiments showed better performances of Ca-P coated surface by IBAD method with respect to Ca-P coated surface by Sol-gel process. Ca-P coated surface by IBAD method appear to give rise more mature osteoblast characteristics and might result in increased bone growth and bone-implant contact.
Porous ${\beta}-tricalcium$ phosphate $({\beta}-TCP)$ bioceramic was fabricated by pressureless sintering using commercial HAp and different volume percentages of PMMA powders (30-60 vol.%). The range of spherical pore size was about $200-250\;{\mu}m$ in diameter. By increasing the PMMA content, the number of pores and their morphology were dramatically changed as well as decreased the material properties. In case of using 60 vol.% PMMA content, network-type pores were found, due to the necking of the PMMA powders. The values of relative density, elastic modulus, bending strength and hardness of the 60 vol.% PMMA content sample, sintered at $1500^{\circ}C$, were about 46%, 22.2 GPa, 5MPa and 182 Hv respectively. Human osteoblast-like MG-63 cells and osteoclast-like Raw 264.7 cells were well grown and fully covered all of the porous ${\beta}-TCP$ bodies sintered at $1500^{\circ}C$.
Kim, Ki-Ho;Kim, Young-Hee;Song, Ho-Yeon;Lee, Byong-Taek
Journal of the Korean Ceramic Society
/
v.43
no.8
s.291
/
pp.504-508
/
2006
Functionally gradient porous $Al_2O_3-(t-ZrO_2)/HAp$ composites consist of 3 layers were fabricated using the multi-pass extrusion process at the various temperatures. The continuous pores were homogeneously formed in the $2^{nd}$ passed samples and their size was about $180{\mu}m$ in diameter. In the porous composites sintered at $1200-1400^{\circ}C$, the relative density and bending strength increased with the sintering temperature. The maximum values of relative density and bending strength in the $2^{nd}$ passed $Al_2O_3-(t-ZrO_2)/HAp$ composites were 62.2% and 107.8 MPa, respectively. In order to investigate the growth behavior of osteogenic cells on the functionally gradient porous $Al_2O_3-(t-ZrO_2)/HAp$ composites, an in vitro test was performed, using human osteoblast-like MG-63 cells. The cells were well attached and grown on the rough surface of the inside of the functionally gradient porous body.
Jang, Won Seok;Kim, Min Gu;Hwang, Dae Suk;Kim, Gyoo Cheon;Kim, Uk Kyu
International Journal of Oral Biology
/
v.42
no.4
/
pp.203-211
/
2017
The aim of this study was to evaluate the role of demineralized and particulate autogenous tooth, and interleukin-6 in bone regeneration. A demineralized and particulate autogenous tooth was prepared and human osteoblast-like cells (MG63) and human osteosarcoma cells were inoculated into the culture. The rate of cell adhesion, proliferation and mineralization were examined, and the appearance of cellular attachment was observed. An 8 mm critical size defect was created in the cranium of rabbits. Nine rabbits were divided into three groups including: An experimental group A (3 rabbits), in which a demineralised and particulate autogenous tooth was grafted; an experimental group B (3 rabbits), in which a demineralized, particulate autogenous tooth was grafted in addition to interleukin-6 (20 ng/mL); and a control group. The rabbits were sacrificed at 1, 2, 4 and 6 weeks for histopathological examination with H-E and Masson's Trichrome, and immunohistochemistry with osteocalcin. The cell-based assay showed a higher rate of cell adhesion, mineralization and cellular attachment in the experimental group A compared with the control group. The animal study revealed an increased number of osteoclasts, newly formed and mature bones in the experimental group A compared with the control group. Eventually, a higher number of osteoclasts were observed in the experimental group B. However, the emergence of newly formed and mature bone was lower than in the experimental group A. The current results suggest that treatment with demineralized and particulate autogenous tooth and interleukin-6 is not effective in stimulating bone regeneration during the bone grafting procedure.
Seo, Bo-Yong;Kim, Young-Min;Choi, Jae-Won;Yun, Mi-Jung;Jeon, Young-Chan;Jeong, Chang-Mo;Kim, Gyu-Cheon;Huh, Jung-Bo
The Journal of Korean Academy of Prosthodontics
/
v.53
no.1
/
pp.9-18
/
2015
Purpose: The purpose of this study is to examine characteristics of implant surface with RBM and anodizing treatments, and to evaluate the responses of osteoblast-like cell (MG-63 cell). Materials and methods: Grade IV titanium disks were fabricated (Diameter 10 mm, thickness 3 mm). Anodizing treatment (ASD) group, RBM and anodizing treatment (RBM/ASD) group, control (machined surface) group were divided. In this study, osteoblast-like cell was used for experiments. The experiments consist of surface characteristics evaluation by FE-SEM images, energy dispersive spectroscopy and stereo-SEM. In order to evaluate cell adhesion evaluation by crystal violet assay and observe cells form by confocal laser microscopy. To assess cell proliferation by XTT assay, cell differentiation by RT-PCR and mineralization by Alizarin red S stain assay. ELISA analyzer was used for Quantitative evaluation. Comparative analysis was run by one-way ANOVA (SPSS version 18.0). Differences were considered statistically significant at P<.05. Results: In ASD group and RBM/ASD group, the surface shape of the crater was observed and components of oxygen and phosphate ions in comparison with the control group were detected. The surface average roughness was obtained $0.08{\pm}0.04{\mu}m$ in the control group, $0.52{\pm}0.14{\mu}m$ in ASD group and $1.45{\pm}0.25{\mu}m$ in RBM/ASD group. In cell response experiments, ASD group and RBM/ASD group were significantly higher values than control group in cell adhesion and mineralization phase, control group was the highest values in the proliferative phase. In RT-PCR experiments, RBM/ASD group was showed higher ALP activity than other groups. RBM/ASD group in comparison with ASD group was significantly higher value for cell adhesion and proliferation phase. Conclusion: In the limitation of this study, we are concluded that the surface treatment with RBM/ASD seems more effective than ASD alone or machined surface on cellular response.
Hydroxyapatite(HA) has been extensively used as bone graft materials and tooth implant surface coating materials because of its biocompatibility and osteoconductive properties. However, as HA is intrinsically poor in mechanical properties, zirconia($ZrO_2$) was incorporated with HA as reinforcing phases for improvement of mechanical properties. The purpose of this study was to investigate the biological activities of HA-coated zirconia through the cell proliferation test, measurements of alkaline phosphatase activity, and histologic examination. Four kinds of tested blocks were prepared according to the pore size (300-500${\mu}m$/500-700${\mu}m$) and the porosity (70%/90%). Cell proliferation and alkaline phosphatase activity was measured at 1, 7, 14 days. The number of cells proliferate after 7, 14 days were significantly increased in all groups when compared with that of the first day, but there was no significant difference between the 4 groups at each time period. At the 7 day, alkaline phosphatase activities of cells cultured in 4 groups were higher than that of the first day, but there was no significant difference between the 4 groups at each time period. The human gingival fibroblast and MG 63 cell was used to evaluate the cell cytotoxicity using MTT test. The materials tested in the current study turned out to be non-cytotoxic. In histologic examination(SEM), at 1 day there were many cells attached on the surfaces of all kinds of tested blocks. The number of cells were increased over time. At the 14 day, there were more cells proliferated than 1 day and some of the pores of blocks were partially filled with the proliferated cells. The in vitro response of osteoblast-like cells to the HA-coated zirconia showed comparable effect on transformation comparable to hydroxyapatite.
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