Yanggu, South Korea
Yanggu, South Korea

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Kim B.-S.,Wonkwang University | Kim B.-S.,Bonecell Biotech Inc. | Kim J.S.,Wonkwang University | Kim J.S.,Bonecell Biotech Inc. | And 4 more authors.
Materials Science and Engineering C | Year: 2013

Magnesium (Mg) is one of the most important ions associated with bone osseointegration. The aim of this study was to evaluate the cellular effects of Mg implantation in titanium (Ti) surfaces treated with sand blast using large grit and acid etching (SLA). Mg ions were implanted into the surface via vacuum arc source ion implantation. The surface morphology, chemical properties, and the amount of Mg ion release were evaluated by scanning electron microscopy (SEM), Auger electron spectroscopy (AES), Rutherford backscattering spectroscopy (RBS), and inductively coupled plasma-optical emission spectrometer (ICP-OES). Human mesenchymal stem cells (hMSCs) were used to evaluate cellular parameters such as proliferation, cytotoxicity, and adhesion morphology by MTS assay, live/dead assay, and SEM. Furthermore, osteoblast differentiation was determined on the basis of alkaline phosphatase (ALP) activity and the degree of calcium accumulation. In the Mg ion-implanted disk, 2.3 × 1016 ions/cm2 was retained. However, after Mg ion implantation, the surface morphology did not change. Implanted Mg ions were rapidly released during the first 7 days in vitro. The MTS assay, live/dead assay, and SEM demonstrated increased cell attachment and growth on the Mg ion-implanted surface. In particular, Mg ion implantation increased the initial cell adhesion, and in an osteoblast differentiation assay, ALP activity and calcium accumulation. These findings suggest that Mg ion implantation using the plasma source ion implantation (PSII) technique may be useful for SLA-treated Ti dental implants to improve their osseointegration capacity. © 2012 Elsevier B.V. All Rights Reserved.


Kim B.-S.,Wonkwang University | Kim B.-S.,Bonecell Biotech Inc. | Park K.E.,Chungnam National University | Park W.H.,Chungnam National University | Lee J.,Wonkwang University
Biomedical Materials (Bristol) | Year: 2013

Electrospinning is a useful method for the production of nanofibrous scaffolds in the field of tissue engineering. Keratin has been used as a biomaterial for electrospinning and can be used in a variety of biomedical applications because it is a natural protein, giving it the ability to improve cell affinity of scaffolds. In this study, keratin was extracted from hagfish slime thread (H-keratin) and blended with polylactic acid (PLA) polymer solution to construct a nanofibrous scaffold. Wool keratin (W-keratin) was used as a control for the comparison of morphological, physical, and biological properties. The results of Fourier transform infrared spectroscopy showed the presence of both W-keratin and H-keratin in the electrospun PLA/keratin. Observations with a scanning electron microscope revealed that PLA, PLA/W-keratin, and PLA/H-keratin had similar average diameters (∼800 nm). Cell attachment experiments showed that MG-63 cells adhered more rapidly and spread better onto PLA/H-keratin than onto the pure PLA or PLA/W-keratin. Cell proliferation assay, DNA content, live/dead, and alkaline phosphatase activity assays showed that PLA/H-keratin scaffolds could accelerate the viability, proliferation, and osteogenesis of MG-63 cells relative to pure PLA or PLA/W-keratin nanofibrous scaffolds. These findings suggest that H-keratin can improve cellular attraction and has great potential to be used as a biomaterial in bone tissue engineering. © 2013 IOP Publishing Ltd.


Kim B.S.,Wonkwang University | Kim B.S.,Bonecell Biotech Inc. | Kang H.-J.,Wonkwang University | Park J.-Y.,Bonecell Biotech Inc. | And 2 more authors.
Experimental and Molecular Medicine | Year: 2015

Fucoidan has attracted attention as a potential drug because of its biological activities, which include osteogenesis. However, the molecular mechanisms involved in the osteogenic activity of fucoidan in human alveolar bone marrow-derived mesenchymal stem cells (hABM-MSCs) remain largely unknown. We investigated the action of fucoidan on osteoblast differentiation in hABMMSCs and its impact on signaling pathways. Its effect on proliferation was determined using the crystal violet staining assay. Osteoblast differentiation was evaluated based on alkaline phosphatase (ALP) activity and the mRNA expression of multiple osteoblast markers. Calcium accumulation was determined by Alizarin red S staining. We found that fucoidan induced hABMMSC proliferation. It also significantly increased ALP activity, calcium accumulation and the expression of osteoblast-specific genes, such as ALP, runt-related transcription factor 2, type I collagen-α 1 and osteocalcin. Moreover, fucoidan induced the expression of bone morphogenetic protein 2 (BMP2) and stimulated the activation of extracellular signal-related kinase (ERK), c-Jun N-terminal kinase (JNK) and p38 mitogen-activated protein kinase by increasing phosphorylation. However, the effect of fucoidan on osteogenic differentiation was inhibited by specific inhibitors of ERK (PD98059) and JNK (SP600125) but not p38 (SB203580). Fucoidan enhanced BMP2 expression and Smad 1/5/8, ERK and JNK phosphorylation. Moreover, the effect of fucoidan on osteoblast differentiation was diminished by BMP2 knockdown. These results indicate that fucoidan induces osteoblast differentiation through BMP2-Smad 1/5/8 signaling by activating ERK and JNK, elucidating the molecular basis of the osteogenic effects of fucoidan in hABM-MSCs. © 2015 KSBMB. All rights reserved 2092-6413/15.


Kim B.-S.,Wonkwang University | Kim B.-S.,Bonecell Biotech Inc. | Kim J.S.,Bonecell Biotech Inc. | Lee J.,Wonkwang University | Lee J.,Bonecell Biotech Inc.
Journal of Biomedical Materials Research - Part A | Year: 2013

Collagen sponges (COL-S) are used as scaffolds to support osteoblasts and stimulate bone repair because of their flexibility, biocompatibility, and biodegradability. In this study, we added fibrin networks to COL-S scaffolds by using a fibrinogen (FNG) cross-linking reaction and evaluated the proliferation, differentiation, and adhesion of MG-63 cells on these scaffolds. The fibrin network that formed in COL-S with various concentrations of FNG was characterized with regard to morphology, porosity, and water-uptake ability. Successful fibrin network formation was observed by scanning electron microscopy (SEM). As the FNG concentration increased, network formation increased, but porosity and water-uptake ability were slightly reduced at high FNG concentrations. An MTS assay, DNA content assay, live/dead fluorescence assay, and SEM imaging showed that MG-63 cells attached and spread on COL-S and COL-S/FNG scaffolds, particularly on scaffolds modified using FNG. In addition, alkaline phosphatase (ALP) activity was significantly increased in cells cultured on scaffolds modified using 10, 40, and 80 mg/mL FNG. Thus, the addition of a fibrin network could increase the biocompatibility of COL-S for bone regeneration. © 2013 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 101A: 2661-2666, 2013. Copyright © 2013 Wiley Periodicals, Inc.


Kim B.-S.,Wonkwang University | Kim B.-S.,Bonecell Biotech Inc. | Kang H.J.,Bonecell Biotech Inc. | Lee J.,Wonkwang University | Lee J.,Bonecell Biotech Inc.
Journal of Biomedical Materials Research - Part B Applied Biomaterials | Year: 2013

Cuttlefish bones (CBs) have emerged as attractive biomaterials because of their porous structure and components that can be converted into hydroxyapatite (HAp) via a hydrothermal reaction. However, their brittleness and low strength restrict their application in bone tissue engineering. Therefore, to improve the compressive strength of the scaffold following hydrothermal conversion to a HAp form of CB (CB-HAp), the scaffold was coated using a polycaprolactone (PCL) polymer at various concentrations. In this study, raw CB was successfully converted into HAp via a hydrothermal reaction. We then evaluated their surface properties and composition by scanning electron microscopy and X-ray diffraction analysis. The CB-HAp coated with PCL showed improved compressive performance and retained a microporous structure. The compressive strength was significantly increased upon coating with 5 and 10% PCL, by 2.09- and 3.30-fold, respectively, as compared with uncoated CB-HAp. However, coating with 10% PCL resulted in a reduction in porosity. Furthermore, an in vitro biological evaluation demonstrated that MG-63 cells adhered well, proliferated and were able to be differentiated on the PCL-coated CB-HAp scaffold, which was noncytotoxic. These results suggest that a simple coating method is useful to improve the compressive strength of CB-HAp for bone tissue engineering applications. © 2013 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater 101B: 1302-1309, 2013. Copyright © 2013 Wiley Periodicals, Inc.


Kim B.S.,Wonkwang University | Kim B.S.,Bonecell Biotech Inc. | Park J.-Y.,Bonecell Biotech Inc. | Kang H.-J.,Wonkwang University | And 3 more authors.
Biochemical and Biophysical Research Communications | Year: 2014

Angiogenesis is an important biological process in tissue development and repair. Fucoidan has previously been shown to potentiate in vitro tube formation in the presence of basic fibroblast growth factor (FGF-2). However, the underlying molecular mechanism remains largely unknown. This study was designed to investigate the action of fucoidan in angiogenesis in human umbilical vein endothelial cells (HUVECs) and to explore fucoidan-signalling pathways. First, we evaluated the effect of fucoidan on cell proliferation. Matrigel-based tube formation and wound healing assays were performed to investigate angiogenesis. Matrix metalloproteinase-2 (MMP-2) mRNA expression and activity levels were analysed by reverse transcription polymerase chain reaction (RT-PCR) and zymography, respectively. Additionally, phosphorylation of mitogen-Activated protein kinases (MAPKs) and protein kinase B (AKT) was detected by Western blot. The results indicate that fucoidan treatment significantly increased cell proliferation in the presence of FGF-2. Moreover, compared to the effect of FGF-2 alone, fucoidan and FGF-2 had a greater effect on tube formation and cell migration, and this effect was found to be synergistic. Furthermore, fucoidan enhanced the phosphorylation of extracellular signal-regulated kinase (ERK), c-Jun N-terminal kinase (JNK), p38, and AKT. MMP-2 activation was also significantly increased. Specific inhibitors of p38 (SB203580) and JNK (SP600125) inhibited tube formation and wound healing, while an ERK inhibitor (PD98059) did not. MMP-2 activation and AKT phosphorylation were also attenuated and associated with the suppression of p38 and JNK phosphorylation, but not with that of ERK. These results indicate that fucoidan, in the presence of FGF-2, induces angiogenesis through AKT/MMP-2 signalling by activating p38 and JNK. These findings provide basic molecular information on the effect of fucoidan on angiogenesis in the presence of FGF-2. © 2014 Elsevier Inc. All rights reserved.


Kim B.-S.,Wonkwang University | Kim B.-S.,Bonecell Biotech Inc. | Yang S.-S.,Wonkwang University | Lee J.,Wonkwang University
Journal of Biomedical Materials Research - Part B Applied Biomaterials | Year: 2014

Cuttlefish bone (CB) is an attractive natural biomaterial source to obtain hydroxyapatite (HAp). In this study, a porous polycaprolactone (PCL) scaffold incorporating CB-derived HAp (CB-HAp) powder was fabricated using the solvent casting and particulate leaching method. The presence of CB-HAp in PCL/CB-HAp scaffold was confirmed by X-ray diffraction (XRD). Scanning electron microscopy (SEM) and porosity analysis showed that the average pore dimension of the fabricated scaffold was approximately 200-300 μm, with ∼85% porosity, and that the compressive modulus increased after addition of CB-HAp powders. In vitro tests such as cell proliferation assay, cytotoxicity analysis, cell attachment observations, and alkaline phosphatase activity assays showed that the PCL/CB-HAp scaffold could improve the proliferation, viability, adherence, and osteoblast differentiation rate of MG-63 cells. When surgically implanted into rabbit calvarial bone defects, consistent with the in vitro results, PCL/CB-HAp scaffold implantation resulted in significantly higher new bone formation than did implantation of PCL alone. These findings suggest that addition of CB-HAp powder to the PCL scaffold can improve cellular response and that the PCL/CB-HAp composite scaffold has great potential for use in bone tissue engineering. © 2013 Wiley Periodicals, Inc.


Cho Y.S.,Wonkwang University | Kim B.-S.,Wonkwang University | Kim B.-S.,Bonecell Biotech Inc. | You H.-K.,Wonkwang University | Cho Y.-S.,Wonkwang University
Current Applied Physics | Year: 2014

In this study, we proposed a novel salt-leaching method using PCL and NaCl powders, known as the SLUP (salt leaching using powder) technique, which has several advantages: this technique does not require solvent, pressure, or unnecessary expensive devices. First, PCL powder (100-180 μm size) and NaCl powder (350-400 μm size) were prepared. Second, the PCL and NaCl powders were mixed at a certain ratio, and then the mixed powder was poured into a mold. Afterward, the mold was heated to melt the PCL powder in an oven at 80 C for 15 min. Subsequently, after the PCL/NaCl mixture was separated from the mold, the PCL/NaCl mixture was soaked in D.I. water for 24 h to leach out the NaCl particles. Consequently, the remaining PCL structure was porous and could be used as a scaffold. To analyze the compressive modulus of the fabricated scaffold, a uniaxial compression test was performed using a UTM (universal testing machine), and the surface characteristics of the scaffold were observed using an SEM (scanning electron microscope). Additionally, cell-culture experiments were performed using hMSCs (human mesenchymal stem cells), and the cell-culture characteristics were assessed and compared with the characteristics from a conventional salt-leaching scaffold. © 2013 Elsevier B.V. All rights reserved.


Kim B.-S.,Bonecell Biotech Inc. | Choi M.-K.,Wonkwang University | Yoon J.-H.,Wonkwang University | Lee J.,Wonkwang University
Oral Surgery, Oral Medicine, Oral Pathology and Oral Radiology | Year: 2015

Objective The aim of this study was to evaluate the in vivo osteogenic potential of biphasic calcium phosphate (BCP), bone morphogenetic protein 2 (BMP-2), and/or mesenchymal stem cell (MSC) composites by using a rabbit calvarial defect model. Study Design Bone formation was assessed by using three different kinds of implants in rabbit calvarial defects, BCP alone, BCP/recombinant human (rh) BMP-2, and BCP/rhBMP-2/MSCs composite. The implants were harvested after 2 or 8 weeks, and the area of new bone formation was quantified by micro-computed tomography (micro-CT) and histologic studies. Results The highest bone formation was achieved with the BCP/rhBMP-2/MSCs treatment, and it was significantly higher than that achieved with the empty or BCP-alone treatment. The quantity of new bone at 8 weeks was greater than at 4 weeks in each group. The relative density of osteocalcin immunoreactivity also increased during this interval. Conclusions These results indicate that the combination of BCP, rhBMP-2, and MSCs synergistically enhances osteogenic potential during the early healing period and could be used as a bone graft substitute. © 2015 Elsevier Inc. All rights reserved.


Kim B.-S.,Bonecell Biotech Inc. | Sung H.-M.,Bonecell Biotech Inc. | You H.-K.,Wonkwang University | Lee J.,Wonkwang University
Journal of Bioscience and Bioengineering | Year: 2014

Fibrin polymers are widely used in the tissue engineering field as biomaterials. Although numerous researchers have studied the fabrication of scaffolds using fibrin glue (FG) and bone powder, the effects of varied fibrinogen content during the fabrication of scaffolds on human mesenchymal stem cells (hMSCs) and bone regeneration remain poorly understood. In this study, we formulated scaffolds using demineralized bone powder and various fibrinogen concentrations and analyzed the microstructure and mechanical properties. Cell proliferation, cell viability, and osteoblast differentiation assays were performed. The ability of the scaffold to enhance bone regeneration was evaluated using a rabbit calvarial defect model. Micro-computed tomography (micro-CT) showed that bone powders were uniformly distributed on the scaffolds, and scanning electron microscopy (SEM) showed that the fibrin networks and flattened fibrin layers connected adjacent bone powder particles. When an 80 mg/mL fibrinogen solution was used to formulate scaffolds, the porosity decreased 41.6 ± 3.6%, while the compressive strength increased 1.16 ± 0.02 Mpa, when compared with the values for the 10 mg/mL fibrinogen solution. Proliferation assays and SEM showed that the scaffolds prepared using higher fibrinogen concentrations supported and enhanced cell adhesion and proliferation. In addition, mRNA expression of alkaline phosphatase and osteocalcin in cells grown on the scaffolds increased with increasing fibrinogen concentration. Micro-CT and histological analysis revealed that newly formed bone was stimulated in the scaffold implantation group. Our results demonstrate that optimization of the fibrinogen content of fibrin glue/bone powder scaffolds will be beneficial for bone tissue engineering. © 2014 The Society for Biotechnology, Japan.

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