Hyun Sook Baek

Evaluation of the Extraction Method for the Cytotoxicity Testing of Latex Gloves

In this study, the cytotoxicity of medical latex gloves to cultured L-929 cells was determined using various extraction conditions. According to the extraction time and temperature, three types of extraction conditions were used: 1) 24 h at 37℃; 2) 72 h at 37℃; 3) 72 h at 50℃. Also, four different extraction vehicles were used, namely, distilled water (DW), 9 g/l sodium chloride (saline) in DW, and culture media with or without serum. Under the above-mentioned conditions, the samples were extracted and then 2-fold serially diluted in the concentration range 3.13 - 50%. When extracted with either DW or saline for 24 h or 72 h at 37℃, only 50% diluted samples showed distinct cytotoxicity to L-929 cells. Moreover, no cytotoxic potentials were observed when gloves were extracted with DW or saline at 50℃ for 72 h. Cytotoxicity was markedly greater when gloves were extracted with culture medium, irrespective of the presence of serum in the medium. These results suggest that optimal extraction conditions should be established for the cytotoxicity evaluations of biomaterials and medical devices.

Biological Advantages of Porous Hydroxyapatite Scaffold Made by Solid Freeform Fabrication for Bone Tissue Regeneration

Presently, commercially available porous bone substitutes are manufactured by the sacrificial template method, direct foaming method, and polymer replication method (PRM). However, current manufacturing methods provide only the simplest form of the bone scaffold and cannot easily control pore size. Recent developments in medical imaging technology, computer-aided design, and solid freeform fabrication (SFF), have made it possible to accurately produce porous synthetic bone scaffolds to fit the defected bone shape. Porous scaffolds were fabricated by SFF and PRM for a comparison of physical and mechanical properties of scaffold. The suggested three-dimensional model has interconnected cubic pores of 500 μm and its calculated porosity is 25%. Whereas hydroxyapatite scaffolds fabricated by SFF had connective macropores, those by PRM formed a closed pore external surface with internally interconnected pores. SFF was supposed to be a proper method for fabricating an interconnected macroporous network. Biocompatibility was confirmed by testing the cytotoxicity, hemolysis, irritation, sensitization, and implantation. In summary, the aim was to verify the safety and efficacy of the scaffolds by biomechanical and biological tests with the hope that this research could promote the feasibility of using the scaffolds as a bone substitute.

Enhanced chondrogenic responses of human articular chondrocytes onto silk fibroin/wool keratose scaffolds treated with microwave-induced argon plasma.

Silk fibroin (SF) is a natural, degradable, fibrous protein that is biocompatible, is easily processed, and possesses unique mechanical properties. Another natural material, wool keratose (WK), is a soluble derivative of wool keratin, containing amino acid sequences that induce cell adhesion. Here, we blended SF and WK to improve the poor electrospinability of WK and increase the adhesiveness of SF. We hypothesized that microwave-induced argon plasma treatment would improve chondrogenic cell growth and cartilage-specific extracellular matrix formation on a three-dimensional SF/WK scaffold. After argon plasma treatment, static water contact angle measurement revealed increased hydrophilicity of the SF/WK scaffold, and scanning electron microscopy showed that treated SF/WK scaffolds had deeper and more cylindrical pores than nontreated scaffolds. Attachment and proliferation of neonatal human knee articular chondrocytes on treated SF/WK scaffolds increased significantly, followed by increased glycosaminoglycan synthesis. Our results suggest that microwave-induced, plasma-treated SF/WK scaffolds have potential in cartilage tissue engineering.

Protection of rabbit kidney from ischemia/reperfusion injury by green tea polyphenol pretreatment

Reactive oxygen species (ROS) have been implicated in the pathogenesis of renal injury after ischemia/reperfusion (I/R). Recently, green tea polyphenols (GTP) have been found to protect the myocardium and liver against I/R injury. Less attention, however, has been paid to the protective effects of GTP with respect to the kidneys. This study was designed to determine whether GTP could protect renal cells from ischemic injury. The rabbits were divided into three groups of equal size: control (sham-operated), I/R + vehicle (normal saline) and I/R + GTP groups. Each group consisted of six rabbits. Animals underwent 30, 60, 90 and 120 min of ischemia, followed by 24 h of reperfusion, respectively. GTP (200 μg/kg) or the vehicle was administered 45 min prior to commencement of I/R. The results demonstrated that GTP administration resulted in a significant (P < 0.05) reduction of renal damage after 90 min of ischemia, as indicated by the decreased levels of creatinine and urea nitrogen in serum. These results were confirmed by histological examinations, which showed that GTP pretreatment inhibited necrosis and sloughing of the proximal tubules induced by I/R. Examinations also showed decreased necrotic areas in the medulla and decreased glomerular collapse in the I/R-injured rabbits. Moreover, the infiltration of CD8+ T cells was considerably decreased in GTP-treated kidneys. The results of this study suggest that GTP can reduce renal injury by preventing the oxidative stress dependent on I/R and may be used in renal transplantation as an antioxidant.

Beneficial effects of microwave-induced argon plasma treatment on cellular behaviors of articular chondrocytes onto nanofibrous silk fibroin mesh

Silk fibroin scaffolds were examined as a biomaterial option for tissue-engineered cartilage-like tissue. In tissue engineering for cartilage repair using a scaffold, initial chondrocyte-material interactions are important for the following cell behaviors. In this study, the surface of nanofibrous silk fibroin (NSF) meshes was modified by a microwave-induced argon plasma treatment in order to improve the cytocompatibility of the meshes used as cartilaginous grafts. In addition, the effects of a plasma treatment on the cellular behavior of chondrocytes on NSF were examined. The plasma treatment resulted in an increase in the hydrophilicity of NSF meshes suggesting that the cytocompatibility of the mesh might be improved. Furthermore, the human articular chondrocytes showed higher viability on the surface-modified NSF meshes. These results suggest that the surface modification of NSF meshes by plasma can enhance the cellular behavior of chondrocytes and may be used in tissue engineering.

Increasing potential risks of contamination from repetitive use of endoscope

BACKGROUND Medical devices inserted into the human body can be divided into single-use devices and reusable medical devices (RMDs) depending on the method of use. RMDs are applied to >1 patient, and users reprocess them prior to repeat use. Therefore, reprocessing of RMDs has become more difficult, and issues related to infection risk have become more prevalent. METHODS Research was performed to compare differences in surface alterations between not-aged (NA) samples and accelerated-aging (AA) samples. NA and AA samples were exposed to potential contaminants and treated with the same cleaning conditions. Then the residual contaminants were analyzed, and adhesion characteristics were investigated. RESULTS Significant differences between the 2 sample groups were observed in the pattern of physical surface alterations. Similar to the endoscope for clinical use, a deep crack was found in the surface of the AA sample. According to the binding affinity test, highly likely potential contaminants were found more frequently on AA samples. CONCLUSION The results suggest the necessity of limiting the duration of time that RMDs may be used, considering the potential risk of infection from repetitive use of RMDs, and also raised the possibility of applying this simulation model of AA. This methodology can be applied to various RMDs, including endoscopes.

Protection of Human Fibroblasts from Reactive Oxygen Species by Green Tea Polyphenolic Compounds

The potential protective roles played by green tea compounds (GTPCs) against reactive oxygen species-induced oxidative stress in cultured fetal human dermal fibroblasts (fHDFs) were investigated according to cell viability measurement methods, such as fluorescence double staining followed by flow cytometry (FCM), MTT assay and crystal violet uptake. Oxidative stress was induced in the fHDFs, either by adding 50 mM H2O2 or by the action of 40 U/L xanthine oxidase (XO) in the presence of xanthine (250 µM). FCM analysis was the most suitable to show that both treatments produced a significant (p < 0.05) reduction in the fHDF viability, attributed to its high sensitivity. On the microscopic observations, the cell death with necrotic morphology was appreciably induced by both treatments. These oxidative stress-induced damages were significantly (p < 0.05) prevented by pre-incubating the fHDFs with 200 µg/ml GTPC for 1 h. These results suggest that GTPC can act as a biological antioxidant in a cell culture experimental model and prevent oxidative stress-induced cytotoxicity in cells.

The Influences of β-Glucan Associated with BMP-7 on MC3T3-E1 Proliferation and Osteogenic Differentiation

b-glucan, an immunomodulator, can selectively enhance the immunobiological activities of neutrophils and macrophages without stimulating proinflammatory cytokine production. Biologic response modifiers, like beta-glucan, will modulate immunity, modify neoplastic disease and increase resistance to microbial challenge. Therefore, beta-glucan polymers can be applied in bone induction and regeneration model and have a possibility of association with bone morphogenetic protein (BMP) because of tissue-regenerative and antimicrobial effects of those polymers. In this report, we studied an E. coli expression system for BMP-7 production and the biological activities of b-glucan associated with BMP-7. The proliferation of MC3T3-E1 osteoblastic cells was enhanced by treatment with Aureobasidium b-glucan, while neither mushroom b-glucan nor barley b-glucan increased the cell proliferation. Mushroom b-glucan alone or associated with BMP-7 increased alkaline phosphatase (ALP) activity of MC3T3-E1 cells, one of the osteoblast phenotype markers, but the other b-glucans did not affect ALP activity of the cells. In mineralization assay, a highly significant increase in nodular staining was observed in cultures treated with both mushroom and Aureobasidium b-glucans in the presence of BMP-7 compared with nontreated controls, while barley b-glucan showed a significant decrease in nodule number compared with cultures treated only with BMP-7.

Growth of Smooth Muscle Cell and Endothelial Cell on PLGA Film containing EGCG

Poly (D,L-latic-co-glycolic acid) (PLGA) has been used as the artificial scaffold for blood vessel formation. In order to hinder smooth muscle cell (SMC) angiogenesis, new scaffold design method of loading Epigallocatechin 3-O-gallate (EGCG) on PLGA film was introduced. PLGA and EGCG were dissolved in acetone and film-shape scaffold was manufactured. Antiangiogenetic effect of EGCG released on scaffold was analyzed for SMC and human umbilical vein endothelial cell (HUVEC) and method for selective inhibition from the difference of growth of SMC and HUVEC was suggested.

Effects Of (1→3), (1→6)-β-D-Glucan Behavior in Human Dermal Fibroblast Cells under Serum Starvation

Glucans have been reported to stimulate immunity and to promote wound healing. Adult human dermal fibroblast (aHDF) cultured in serum free (serum-starvation). Proliferation of aHDF was measured at various concentrations of β-glucan by MTT assay, and migration was observed for 36h on microscope. The result of fibroblast bioassay, β-glucan had positive influence. In this study, the direct effects of β-glucan on proliferation and migration of human dermal fibroblasts were examined in vitro. That means β-D-glucan has the effect to enhance proliferation and aHDF migration speed, and has the potential as a wound healing agent.