• Journal of Microbiology and Biotechnology
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• v.17 no.9
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• pp.1477-1482
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• 2007

Gluconacetobacter diazotrophicus strain PA15 exhibited a minimum inhibitory concentration value of 11 mM in an LGI medium amended with $ZnCl_2$. When an LGI medium was amended with Zn metal, solubilization halos were observed in a plate assay, and further solubilization was confirmed in a broth assay. The maximum solubilization was recorded after 120 h with a 0.1% Zn metal amendment. During solubilization, the culture growth and pH of the broth were indirectly correlated. Using a Fourier Transform Infrared Spectroscopy analysis, one of the agents solubilizing the Zn metal was identified as gluconic acid. When the Zn-amended broth was observed under a bright field microscope, long involution cells were observed, and further analysis with Atomic Force Microscopy revealed highly deformed, pleomorphic, aggregate-like cells.

• Journal of Microbiology and Biotechnology
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• v.9 no.1
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• pp.39-43
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• 1999

A flocculent aggregate produced by Lactococcus lactis subsp. lactis in broths containing Tween 80, including MRS broth, had a microscopic structure of intertwined thread-like filaments. The filamentous structure was not elongated bacterial cells, but consisted of an organic solvent-soluble portion and an insoluble solid. L. lactis subsp. lactis grown at $25^{\circ}C$ for 15 days in tryptic soy broth with 0.1% Tween 80 and 1.0% malt extract produced 13 mg/l of flocculent aggregate, which contained 0.84 g/g of organic solvent-soluble component. The organic solvent-soluble part was identified as 10-hydroxyoctadecanoic acid.

• Restorative Dentistry and Endodontics
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• v.35 no.3
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• pp.222-228
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• 2010

The purpose of this study was to compare mineral trioxide aggregate (MTA; Dentsply, Tulsa Dental, Tulsa, OK, USA), which is widely used as root-end filling material, with DiaRoot BioAggregate (DB; Innovative BioCaramix Inc, Vancouver, BC, Canada), newly developed product, by using MG63 osteoblast-like cells. MTA, DB, and Intermediate Restorative Material (IRM; Dentsply Caulk, Milford, DE, USA) were used for root-end filling material while tissue culture plastic was used for control group. Each material was mixed and, the mixtures were left to set for 24 hours. MG63 cells were seeded to each group and then they were cultured for attachment for 4 hours. Following the attachment of cells to the root-end filling material, early cellular response was observed. After another 12 hours'culture, the level of attachment between cells and material was observed and in order to identify the effect of each material to bone formation, transforming growth factor beta1 ($TGF{\beta}1$) and osteocalin (OC) were estimated by using enzyme-linked immunosorbent assay (ELISA), and the amount of alkaline phosphatase (ALP) was also measured. The data were analyzed using one-way ANOVA. As a result, only at OC and the number of cells which were attached to materials, there was no statistical difference between MTA and DB. At other items, there was statistically significant difference in all groups. Although DB has not shown exactly the same cellular response like that of MTA, the number of attached cells shows that biocompatibility of the material and OC indicates bone formation rate. Therefore, if DB is used for root end filling material, it is expected to lead to similar results to MTA.

• Journal of the korean academy of Pediatric Dentistry
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• v.40 no.3
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• pp.185-193
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• 2013

Mineral trioxide aggregate (MTA) has recently been used as a pulpotomy medicament for primary molars. The aim of this study was to evaluate and compare the proliferation and differentiation potential of dental pulp stromal cells of permanent teeth and deciduous teeth cultured on MTA-coated surface. Human dental pulp stromal cells were obtained from human permanent premolars and deciduous teeth and cultured on MTA-coated culture plates. The cells were subjected to proliferation assay and cell cycle analysis. Their differentiation potential was evaluated by analysing changes in the mRNA expressions of runt-related transcriptional factor 2 (Runx2) and alkaline phosphatase (ALP). Morphological changes of cells in direct contact with MTA were observed using scanning electron microscopy (SEM). The proliferation rates, distribution of cell cycles and mRNA expression patterns of Runx2 and ALP were similar in both types of pulpal cells. SEM observations revealed that both types changed into more dendrite-like cells. On the surface of MTA, human dental pulp stromal cells from deciduous and permanent teeth were able to both proliferate and differentiate into cells that induce mineralization. MTA is suitable as a biocompatible pulpotomy medicament for primary teeth.

• International Journal of Oral Biology
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• v.37 no.2
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• pp.77-83
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• 2012

We investigated the pulpal response to direct pulp capping in rat molar teeth using mineral trioxide aggregate (MTA) and calcium hydroxide (CH). A palatal cavity was prepared in rat maxillary molar teeth. Either MTA or CH was placed on the exposed pulp and all cavities were restored with composite. Rats were sacrificed for histological evaluation after 12 hours and at 2, 7, 14 and 21 days. In both the MTA and CH groups, reparative dentin formation was clearly observed on histology after 14 days. The MTA-capped pulps were found to be mostly free from inflammation, and hard tissue of a tubular consistent barrier was observed. In contrast, in CH-capped teeth, excessive formation of reparative dentin toward residual pulp was evident. The pulpal cell response beneath the reparative dentin layer was examined by immunofluorescence using antibodies against DSP. After 2 days, a few DSP immunopositive cells, most of which showed a cuboidal shape, appeared beneath the predentin layer. At 7 days, DSP-immunopositive cells with columnar odontoblast-like cells were seen beneath the newly formed hard tissues. At 14 and 21 days, DSP was more abundant in the vicinity of the odontoblastic process along the dentinal tubules than in the mineralized reparative dentin. The CH group showed strong expression patterns in terms of DSP immunoreactivity. Our results thus indicate that MTA may be a more effective pulp capping material as it induces the differentiation of odontoblast-like cells and the formation of reparative dentin without the loss of residual pulp functions.

• Restorative Dentistry and Endodontics
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• v.35 no.5
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• pp.359-367
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• 2010

Objectives: The purpose of the present in vitro study was to evaluate the biocompatibility of mineral trioxide aggregate (MTA) mixed with glass ionomer cement (GIC), and to compare it with that of MTA, GIC, IRM and SuperEBA. Materials and Methods: Experimental groups were divided into 3 groups such as 1 : 1, 2 : 1, and 1 : 2 groups depending on the mixing ratios of MTA powder and GIC powder. Instead of distilled water, GIC liquid was mixed with the powder. This study was carried out using MG-63 cells derived from human osteosarcoma. They were incubated for 1 day on the surfaces of disc samples and examined by scanning electron microscopy. To evaluate the cytotoxicity of test materials quantitatively, XTT assay was used. The cells were exposed to the extracts and incubated. Cell viability was recorded by measuring the optical density of each test well in reference to controls. Results: The SEM revealed that elongated, dense, and almost confluent cells were observed in the cultures of MTA mixed with GIC, MTA and GIC. On the contrary, cells on the surface of IRM or SuperEBA were round in shape. In XTT assay, cell viability of MTA mixed with GIC group was similar to that of MTA or GIC at all time points. IRM and SuperEBA showed significantly lower cell viability than other groups at all time points (p < 0.05). Conclusions: In this research MTA mixed with GIC showed similar cellular responses as MTA and GIC. It suggests that MTA mixed with GIC has good biocompatibility like MTA and GIC.

• Journal of Microbiology and Biotechnology
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• v.20 no.2
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• pp.325-331
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• 2010

Rhodococcus erythropolis amidase was expressed in Escherichia coli cells. The crude amidase in the cell-free extract was immobilized using the cross-linked enzyme aggregate (CLEA) method. The crude amidase was mixed with bovine serum albumin and then precipitated with ammonium sulfate. The resultant precipitant was subsequently cross-linked with glutaraldehyde. Scanning electron microscopy revealed that this co-CLEA had a ball-like shape with a diameter of approximately $1\;{\mu}m$. This co-CLEA evidenced hydrolytic activity toward a variety of amide substrates. The amidase co-CLEA evidenced an optimum temperature of $60^{\circ}C$ and an optimum pH of 8.0, results that were similar to those of the soluble amidase. The reaction stability of the co-CLEA was increased. That is, it was stable up to $50^{\circ}C$ and in a pH range of 5.0-12.0. Additionally, the co-CLEA could be recovered by centrifugation, and retained 96% activity after 3 repeated cycles. This amidase co-CLEA may prove useful as a substitute for soluble amidase as a biocatalyst in the pharmaceutical and chemical industries.

• Applied Microscopy
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• v.28 no.3
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• pp.363-377
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• 1998

After the investigation on the eye of Incilaria fruhstorieri with light and electron microscopes, the following results were obtained. The eye of Incilaria fruhstorferi comprises cornea, lens, vitreous body, retina, and optic nerve inward from the outside. Cornea is composed of squamous, cuboid, columnar and irregular cells, which appear to be light due to their low electron density. In their cytoplasms, glycogen granules, multivesicular body, and nucleus were observed. Vitreous body, located behind non-cellular transparent lens, is filled with long and short microvilli protruding from the retinal epithelia. Retinal epithelium, the organ to perceive objects, is divided into four parts; microvillar layer pigment layer, nuclear layer, and neutrophils layer, from the apical portion. Microvillar layer consists of the type-I photoreceptor cells and pigmented granule cells. In the apical portion of their cytoplasms, long microvilli (length, $19{\mu}m$) , short microvilli (length, $8{\mu}m$), and rolled microvilli grow thick in the irregular and mixed forms. Photoreceptor cells are classified into type-I and type-II, according to their structures. The type-I cell has the apical portion rising roundly like a fan and the lower part which looks like the helve of a fan. In the cytoplasm of the apical portion, there are clear vesicles, cored vesicles, ovoid mitochondria, and microfilaments, and in the cytoplasm of the lower part, photic vesicles with their diameters about 60nm aggregate densely. The type-II photoreceptor cell, located at the lower end of the type-I cells, has a very large ovoid nucleus 3nd no microvilli. In the cytoplasm of the type-II cell, the photic vesicles with sizes 60nm aggregate more densely than in the cytoplasm of the type-I cell. Pigmented cells are classified into type-A and type-B, according to their structures. The type-A is identified to be a large cell containing round granules (diameter, $0.5{\mu}m$) of very high electron density, while the type-B is identified as a small cell where the irregular granules (diameter, $0.6{\mu}m$) of a little lower electron density amalgamate. Nuclear layer ranges from the bottom of pigment layer to the top of the capsule, and contains three kinds of nuclei (nuclei of the type-II photoreceptor cell, pigmented granule cell, and accessory neuron). The capsules covering the outmost part of the eyeball are composed of collagenous fiber and three longitudinal muscle layers (the thickness of each longitudinal muscle layer, $0.4{\mu}m$) and thick circular muscle layer (thickness, $0.3{\mu}m$). Around the capsules, there is a neurophile layer consisting of neurons and nerve fibers. Each neuron has a relatively large ovoid nucleus for its cytoplasm, and in the karyosome, large lumps of keterochromatin form a wheel nucleus.

• Journal of the korean academy of Pediatric Dentistry
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• v.42 no.4
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• pp.291-301
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• 2015

This study compared the in vitro cell viability and differentiation potentials of human deciduous dental pulp cells (DPCs) on mineral trioxide aggregate (MTA)-like products (ProRoot MTA, RetroMTA and Endocem Zr). The experimental materials were prepared as circular discs, which were used to test the effects of the materials on the viability of human DPCs when placed in direct and indirect contact. Furthermore, the pH of the extracted materials was recorded, and their effect on cell differentiation potential was evaluated by evaluating the alkaline phosphatase (ALP) activity and Alizarin Red S staining of DPCs incubated with the test materials. In direct contact, the cell viability of human DPCs was higher with ProRoot MTA and RetroMTA than with Endocem Zr. However, when in indirect contact, the cell viability of human DPCs was generally higher in Endocem Zr than in ProRoot MTA and Retro MTA. With respect to pH, the alkalinity was lower for Endocem Zr than for the other test materials. The ALP activities of the cells were not enhanced by any of the experimental materials. Alizarin Red S staining of the tested human DPCs revealed that their differentiation potential was lower than for cells incubated with osteogenic induction medium. While there were differences in the responses of the human DPCs to the test materials, all displayed degrees of cytotoxicity and were unable to enhance either the viability or differentiation of human DPCs. However, Endocem Zr exhibited better cell viability and was less alkaline than the other test materials.

• Proceedings of the Korean Society of Developmental Biology Conference
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• 2003.10a
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• pp.100-100
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• 2003

Pluripotent embryonic stem (ES) cells differentiate spontaneously into beating cardiomyocytes via embryo-like aggregates. We describe the use of mouse embryonic stem (mES03) cells as a reproducible differentiation system for cardiomyocyte. To induce cardiomyocytic differentiation, mES03 cells were dissociated and allowed to aggregate (EB formation) at the presence of 0 75% dimethyl sulfoxide (DMSO) for 4 days and then another 4 days without DMSO (4+/4-). Thus treated EBs were plated onto gelatin-coated dish for differentiation. Spontaneously contracting colonies which appeared in approximately 4-5 days upon differentiation. Expression of cardiac-specific genes were determined by RT-PCR. Rebust expression of myosin light chain (MLC-2V), cardiac myosin heavy chain $\alpha$, cardiac muscle heavy polypeptide 7 $\beta(\beta$-MHC), cardiac transcription factor GATA4 and skeletal muscle-specific ${\alpha}_1$-subunit of the L-type calcium channel (${\alpha}_1 CaCh_{sm}$) were detected as early as 8 days after EB formation, but message of cardiac muscle-specific $\alpha$$_1$-subunit of the L-type calcium channel (${\alpha}_1$CaCh) were revealed at a low level. Strikingly, the expression of atrial natriuretic factor (ANF) was not detected. When spontaneous contracting cell masses were examined their electrophysiological features by patch-clamp technique, it showed ventricle-like action potential 17 days after the EB formation. This study indicates that mES03 cell-derived cardiomyocytes displayed biochemical and electrophysiological properties of cardiomyocytes and DMSO enhanced development of cardiomyocytes in 4+/4- method.