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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.


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.


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.,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.

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