• Macromolecular Research
• /
• v.12 no.4
• /
• pp.374-378
• /
• 2004

The biodegradable and biocompatible poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV), a copolymer of microbial polyester, was fabricated as nanofibrous mats by electrospinning. Image analysis of the electrospun nanofibers fabricated from a 2 wt% 2,2,2-trifluoroethanol solution revealed a unimodal distribution pattern of fiber diameters with an observed average diameter of ca. 185 nm. The fiber diameter of electrospun fabrics could be controlled by adjusting the electro spinning parameters, including the solvent composition, concentration, applied voltage, and tip-to-collector distance. Chondrocytes derived from rabbit ear were cultured on a PHBV cast film and an electrospun PHBV nano-fibrous mat. After incubation for 2 h, the percentages of attached chondrocytes on the surfaces of the flat PHBV film and the PHBV nanofibrous mat were 19.0 and 30.1 %, respectively. On the surface of the electrospun PHBV fabric, more chondrocytes were attached and appeared to have a much greater spreaded morphology than did that of the flat PHBV cast film in the early culture stage. The electro spun PHBV nanofabric provides an attractive structure for the attachment and growth of chondrocytes as cell culture surfaces for tissue engineering.

• Microbiology and Biotechnology Letters
• /
• v.17 no.3
• /
• pp.231-235
• /
• 1989

Possibility of using microcarriers for the growth of a transformed human embryonic kidney cell line 293 was investigated. The cell grew well in a static culture such as T-flasks with medium of DME/F12 (3:1) mixture supplemented with 5% FBS, but it was most difficult to make the cells grow on microcarriers mainly due to the low attachment efficiency and poor spreading at initial stage of the culture. Consequently, 30-50% of the cells were lost upon inoculation into microcarrier suspension and significant fraction of the mirrocarrier became bald. The medium supplemented with the concentrated conditioned medium by hepatoma cell line HpG2 supported the active growth of the cells on microcarrier and the cells showed a very healthy and well spreading morphology. It was probable that some spreading and attachment factors of HpG2 conditioned medium were effective for 293 cells.

• BMB Reports
• /
• v.29 no.2
• /
• pp.175-179
• /
• 1996

Viral surface proteins are known to play an essential role in attachment of the virus particle to the host cell membrane. In case of the hepatitis B virus (HBV) several reports have described potential receptors on the target cell side, but no definite receptor protein has been isolated yet. As for the viral side, it has been suggested that the preS region of the envelope protein, especially the preS1 region, is involved in binding of HBV to the host cell. In this study, preS1 region was recombinantly expressed in the form of a maltose binding protein (MBP) fusion protein and used to identify and visualize the expression of putative HBV receptor(s) on the host cell. Using laser scanned confocal microscopy and by FACS analysis, MBP-preS1 proteins were shown to bind to the human hepatoma cell line HepG2 in a receptor-ligand specific manner. The binding kinetic of MBP-preS1 to its cellular receptor was shown to be temperature and time dependent. In cells permeabilized with Triton X-100 and treated with the fusion protein, a specific staining of the nuclear membrane could be observed. To determine the precise location of the receptor binding site within the preS1 region, several short overlapping peptides from this region were synthesized and used in a competition assay. In this way the receptor binding epitope in preS1 was revealed to be amino acid residues 27 to 51, which is in agreement with previous reports. These results confirm the significance of the preS1 region in virus attachment in general, and suggest an internalization pathway mediated by direct attachment of the viral particle to the target cell membrane.

• Journal of Periodontal and Implant Science
• /
• v.41 no.2
• /
• pp.67-72
• /
• 2011

Purpose: The aim of this study is to determine whether certain biomaterials have the potential to support cell attachment. After seeding bone marrow stromal cells onto the biomaterials, we investigated their responses to each material in vitro. Methods: Rat bone marrow derived stromal cells were used. The biomaterials were deproteinized bovine bone mineral (DBBM), DBBM coated with fibronectin (FN), synthetic hydroxyapatite (HA), HA coated with FN, HA coated with $\beta$-tricalcium phosphate (TCP), and pure $\beta$-TCP. With confocal laser scanning microscopy, actin filaments and vinculin were observed after 6, 12, and 24 hours of cell seeding. The morphological features of cells on each biomaterial were observed using scanning electron microscopy at day 1 and 7. Results: The cells on HA/FN and HA spread widely and showed better defined actin cytoskeletons than those on the other biomaterials. At the initial phase, FN seemed to have a favorable effect on cell adhesion. In DBBM, very few cells adhered to the surface. Conclusions: Within the limitations of this study, we can conclude that in contrast with DBBM not supporting cell attachment, HA provided a more favorable environment with respect to cell attachment.

• Fisheries and Aquatic Sciences
• /
• v.18 no.1
• /
• pp.81-88
• /
• 2015

We investigated factors affecting primary cultures of Pacific abalone Haliotis discus hannai ovary-dissociated cells to identify general aspects of their early-phase culture. Ninety-seven cell populations derived from 30 individuals were cultured in different media with varying compositions of medium supplements, and initial attachment, subculture, and survival for ${\geq}10$ weeks were assessed according to medium composition and individual. We also examined the time required for subculture and the rate of cell death according to both culturing period and passage number within 10 weeks. A lack of fetal bovine serum (FBS) and hemolymph significantly inhibited the growth of cultured cells, while we detected no significant effect of medium composition on initial cell attachment. Through data reallocation, with the omission of data from cell populations cultured in FBS-free and hemolymph-free media, we showed that growth inhibition was also affected by individual differences among the abalones used. During the culture, we observed four different types of cell morphology. Moreover, considerable time was required for subculture-18.4 and 19.5 days for first and second subcultures, respectively-and cell death did not occur within 30 days or for passage 0. Our results will provide valuable information for developing universal cell culturing guidelines in abalone species and suggest the feasibility of culturing abalone ovary-dissociated cells.

• Journal of Periodontal and Implant Science
• /
• v.31 no.1
• /
• pp.163-180
• /
• 2001

The aim of this study was to evaluate the effects of chitosan coating on the attachment, proliferation, functional and morphological change of periodontal ligament cells. Primary human periodontal ligament cells were cultured in dulbecco's modified Eagle's medium with 10% fetal bovine serum and 1% antibiotics. In experimental group, cells of 4th to 7th passage were inoculated in the multiwell plates coated with chitosan in concentration of 0.22, 0.2, and $2mg/m{\ell}$. Cell counting and MTT assay were done after 0.5, 1.5, 3, 6 and 24 hours of incubation to evaluate the cell attachment, and then after 2 and 7 days of culture to evaluate the cell proliferation. The alkaline phosphatase activity was measured after 4 and 7 days of culture and the ability to produce mineralized modules was evaluated after 21 days of culture. The results were as follows : 1. The morphology of periodontal ligament cells on the chitosan coating was round or spheric. Round cells were aggregated after 6 hours of culture. Aggregated cells on the chitosan coated surface showed nodule-like appearance after 24 hours of culture and not achieved confluency at 7 days. 2. During early period of culture, the attachment of periodontal ligament cells were inhibited by chitosan coating. Inhibition of cell attachment tended to increase with the concentration of chitosan. 3. At the chitosan concentration of 0.02 and $0.2mg/m{\ell}$, periodontal ligament cells were more rapidly proliferated at 7 days, compared to the control group. At the concentration of $2mg/m{\ell}$, the proliferation of periodontal ligament cells was inhibitied(p<0.01). 4. Alkaline phosphatase activity of periodontal ligament cells was increased in chitosan coated group, especially at the concentration of $0.02mg/m{\ell}$after 4 days of culture.5. Periodontal ligament cells produced mineralized nodules on chitosan coated wells without the addition of mineralized nodule forming materials (ascorbic acid, ${\beta}-glycerophosphat$, dexamethasone). With the addition of mineralized nodule forming materials, periodontal ligament cells produced more mineralized nodules at the concentration of $0.02mg/m{\ell}$, compared to the control. In summary, the attachment, proliferation, cell activity, and alkaline phosphatase activity of periodontal ligament cells depended on the concentration of coated chitosan. Chitosan stimulated mineralized nodule formation by periodontal ligament cells. At the appropriate concentration($0.02mg/m{\ell}$), chitosan could increase alkaline phosphatase activity and stimulate the formation of mineralized nodule by periodontal ligament cells. These results suggest that chitosan can be used as an adjunct for bone graft material, and the matrix of tissue engineering for periodontal regeneration, especially bone regeneration.

• The Journal of Advanced Prosthodontics
• /
• v.6 no.5
• /
• pp.406-414
• /
• 2014

PURPOSE. The objective of this study was to investigate the biologic effects of enamel matrix derivative (EMD) with different concentrations on cell viability and the genetic expression of human gingival fibroblasts (HGF) to zirconia surfaces. MATERIALS AND METHODS. Immortalized human gingival fibroblasts (HGF) were cultured (1) without EMD, (2) with EMD $25{\mu}g/mL$, and (3) with EMD $100{\mu}g/mL$ on zirconia discs. MTT assay was performed to evaluate the cell proliferation activity and SEM was carried out to examine the cellular morphology and attachment. The mRNA expression of collagen type I, osteopontin, fibronectin, and TGF-${\beta}1$ was evaluated with the real-time polymerase chain reaction (RT-PCR). RESULTS. From MTT assay, HGF showed more proliferation in EMD $25{\mu}g/mL$ group than control and EMD $100{\mu}g/mL$ group (P<.05). HGFs showed more flattened cellular morphology on the experimental groups than on the control group after 4h culture and more cellular attachments were observed on EMD $25{\mu}g/mL$ group and EMD $100{\mu}g/mL$ group after 24h culture. After 48h of culture, cellular attachment was similar in all groups. The mRNA expression of type I collagen increased in a concentration dependent manner. The genetic expression of osteopontin, fibronectin, and TGF-${\beta}1$ was increased at EMD $100{\mu}g/mL$. However, the mRNA expression of proteins associated with cellular attachment was decreased at EMD $25{\mu}g/mL$. CONCLUSION. Through this short term culture of HGF on zirconium discs, we conclude that EMD affects the proliferation, attachment, and cell morphology of HGF cells. Also, EMD stimulates production of extracellular matrix collagen, osteopontin, and TGF-${\beta}1$ in high concentration levels. CLINICAL RELEVANCE. With the use of EMD, protective barrier between attached gingiva and transmucosal zirconia abutment may be enhanced leading to final esthetic results with implants.

• Journal of the Korean Society for Marine Environment & Energy
• /
• v.12 no.2
• /
• pp.91-95
• /
• 2009

To understand the effect of attachment substrate on the growth of benthic microalgae, we experimentally investigated the growth of benthic microalgae Nitzschia sp. (Jinhae Bay strain) with additions of glass beads in different sizes. The glass beads used in this study are 0.09-0.15 mm (G1), 0.25-0.50 mm (G2), 0.75-1.00 mm (G3) and 1.25-1.65 mm (G4). No addition of glass beads used as controls. Highest specific growth rate (0.37/day) and maximum cell density ($9,232{\pm}840$ cells/mL) of Nitzschia sp. showed at the smallest glass beads (G1), and the specific growth rate and maximum cell density were decreasing with increasing size of glass beads (specific growth rate and maximum cell density of G4 was 0.24/day and $6,397{\pm}524$ cells/mL, respectively). Moreover, specific growth rate of the control experiment (0.23/day) was significantly lower than their of G1 to G3 experiment. The results indicated that the attachment substrate for benthic microalgae as Nitzschia sp. is important factor which affecting the growth rate as well as cell density. Therefore, the physiological experiment of benthic microalgae seems to be necessary to preliminary experiment, which is addition or not of the attachment suitable substrate and the grain size for the target species of benthic microalgae.

• Asian Journal of Atmospheric Environment
• /
• v.11 no.2
• /
• pp.131-137
• /
• 2017

A cyclone is an effective tool to facilitate the collection of aerosol particles without using filters, and in cell exposure studies is able to collect a sufficient amount of aerosol particles to evaluate their adverse health effect. In this study, we examined two different methods to improve the aerosol particle collection efficiency of a cyclone. The individual and combined effects of reducing the surface roughness of the inner wall of the cyclone and of using a circular cone attachment were tested. The collection efficiency of particles of diameter $0.2{\mu}m$ was improved by approximately 10% when using a cyclone with a smoothened inner wall (average roughness $Ra=0.08{\mu}m$) compared with the original cyclone ($Ra=5.1{\mu}m$). A circular cone attachment placed between the bottom section of the cyclone and the top section of a collection bottle, resulted in improved collection of smaller particles without the attachment. The 50% cutoff diameter of the modified cyclone (combined use of smoothened inner wall and attachment) was $0.23{\mu}m$ compared to $0.28{\mu}m$ in the original model. The combined use of these two techniques resulted in improved collection efficiency of aerosol particles.

• Journal of Applied Biological Chemistry
• /
• v.54 no.3
• /
• pp.166-170
• /
• 2011

Silk fibroin, a natural protein produced by silkworm, is a good biomaterial which has biodegradability and biocompatibility. To ascertain the effects of silk fibroin on cell growth, silk fibroin films were prepared using silk fibroin aqueous solutions of various concentrations. We investigated the attachment, proliferation, morphology of the cells and the expression levels of genes related to cell attachment and growth on the silk fibroin films. When the cells were cultured on the 0.1 and 1% silk fibroin film, the cell adhesion ability was very excellent. Particularly, overall cell growth on the 1% silk fibroin film was definitely superior to the others. Also, expression levels of genes related cell growth were increased on the 0.1 and 1% silk fibroin film. These results suggest silk as a material for medical applications.