• Journal of Periodontal and Implant Science
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• v.37 no.sup2
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• pp.447-463
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• 2007

Periodontal ligament (PDL) cells have been known as multipotential cells, and as playing an important rolesin periodontal regeneration. The PDL cells are composed of heterogeneous cell populations which have the capacity to differentiate into either cementoblasts or osteoblasts, depending on needs and conditions. Therefore, PDL cells have the capacity to produce mineralized nodules in vitro in mineralization medium which include ascorbic acid, ${\beta}$-glycerophosphate and dexamethasone. In spite of these well-known osteoblast like properties of PDL cells, very little is known about the molecules involved in the formation of the mineralized nodules in the PDL cells. In the present study, we analysed gene-expression profiles during the mineralization process of cultured PDL cells by means of a cDNA microarray consisting of 3063 genes. Nodules of mineralized matrix were strongly stained with alizarin red S on the PDL cells cultured in the media with mineralization supplements. Among 3,063 genes analyzed, 35 were up-regulated more than two-fold at one or more time points in cells that developed matrix mineralization nodules, and 38 were down-regulated to less than half their normal level of expression. In accord with the morphological change we observed, several genes related to calcium-related or mineral metabolism were induced in PDL cells during osteogenesis, such as IGF-II and IGFBP-2. Proteogycan 1, fibulin-5, keratin 5, ,${\beta}$-actin, ${\alpha}$-smooth muscle actin and capping protein, and cytoskeleton and extracellular matrix proteins were up-regulated during mineralization. Several genes encoding proteins related to apoptosis weredifferentially expressed in PDL cells cultured in the medium containing mineralization supplements. Dkk-I and Nip3, which are apoptosis-inducing agents, were up-regulated, and Btf and TAXlBP1, which have an anti-apoptosis activity, were down-regulated during mineralization. Also periostin and S100 calciumbinding protein A4 were down-regulated during mineralization.

• Development and Reproduction
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• v.6 no.2
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• pp.97-104
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• 2002

In most mammals, mature oocyte-cumulus complexer(OCCs) ovulate into the oviduct where fertilization by sperm takes place. However, the complex that fail to fertilize eventually undergoes degeneration while they reside in the oviduct. Yet there is no blown mechanism how both oocyte and cumulus cells degenerate. Using human follicular fluid (hFF), bovine oviductal tissue extract (BOX) and mouse OCC, the present study aimed to find how the oviduct influence the viability of the oocyte and cumulus cells in vitro. There was no difference of oocyte maturation rate between the control and BOX-treated groups. However, there was a significant difference in the survival of cumulus cells between two groups. Cumulus cells cultured in the presence of hFF alone underwent initially expansion and then they formed monolayer in the culture dish. Even after 72 hr, they proliferated well and showed fibroblast-like morphology. Cumulus cells cultured in the presence of both hFF and BOX also expanded after 24 hr, however, after 72 hr culture, they eventually detached and degenerated. Cumulus cells cultured in the BOX alone gave a similar drastic result. When the cumulus cells cultured in the presence of BOX were stained with DAPI, their nuclei showed partial condensation and fragmentation. After detailed analysis of these cells by TUNEL assay, many nuclei of them exhibited well stained spots indicating the signs of apoptosis. Based upon these observations, it is suggested that BOX might possess a factor that leads mouse cumulus cells to undergo apoptosis in vitro.

• Tuberculosis and Respiratory Diseases
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• v.50 no.4
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• pp.426-436
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• 2001

Background : Chronic obstructive pulmonary disease(COPD) is one of the major contributors to morbidity and mortality among the adult population. Cigarette smoking(CS) is undoubtedly the single most important factor in the pathogenesis of COPD. However, its mechanism is unclear. The current hypothesis regarding the pathogenesis of COPD postulates that an imbalance between proteases and antiproteases leads to the destructive changes in the lung parenchyma. This study had two aims. First, to evaluate the effect of CS exposure on histologic changes of the lung parenchyme, and second, to evaluate the effect of CS exposure on the expression of the gelatinolytic enzymes in BAL fluid cells in guinea pigs. Methods : Two groups of five guinea pigs were exposed to the whole smoke of 20 commercial cigarettes per day, 5 hours/day, 5 days/week, for 6weeks, and 12 weeks, respectively, using a smoking apparatus. Five age-matched guinea pigs exposed to room air were used as controls. Five or more sections were microscopically extamined(${\times}400$) and the number of cellular infiltration of the alveolar wall was measured in order to evaluate the effect of CS exposure on the histologic changes of lung parenchyme. The statistical significance was analyzed by a linear regression method. To evaluate the expression of the gelatinolytic enzymes in intraalveolar cells, BAL fluid was obtained and the intraalveolar cells were separated by centrifugation (500 g for 10 min at $4^{\circ}C$). Two sets of culture plates were loaded with $1{\times}10^6$ intraalveolar cells. One plate, contained O.1mM EDTA, a inhibitor of matrix metalloproteases(MMPs), and the other plate had no EDTA. Both plates were incubated for 48 hours at $37^{\circ}C$. After incubation, gelatinolytic protease expression in the supernatants was analyzed by gelatin zymography. Results : At the end of CS exposure, the level of blood carboxy Hb had increased significantly(4.1g/dl in control group, 24g/dl immediately after CS exposure, 18g/dl 30 min after CS exposure, 15g/dl 1 hour after CS exposure). Alveolar inflammatory cells were identified in the CS exposed guinea pigs. The number of alveolar cellular cells observed in a microscopic field ($400{\times}$) was $121.4{\pm}7.2$, $158.0{\pm}20.2$, $196.8{\pm}32.8$, in the control, the 6 weeks, and the 12 weeks group, respectively. The increased extent of inflammatory cellular infiltration of the lung parenchema showed a statistically significant linear relationship with the duration of CS exposure(p=0.001, $r^2=0.675$). Several types of gelatinolytic enzymes in the intraalveolar cells of CS exposed guinea pigs were expressed, of which some were inhibited by EDT A. However, the gelatinolytic enzymes were not expressed in the control groups. Conclusion : CS exposure increases inflammatory cellular infiltration of the alveolar wall and the expression of gelatinolytic proteases in guinea pigs. EDTA inhibits some of the gelatinolytic proteases. These findings suggest a possibility that CS exposure may increase MMP expression in the lungs of guinea pigs.

• Applied Chemistry for Engineering
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• v.18 no.3
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• pp.262-266
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• 2007

The effect of the presence of magnesium ions in the preparation of calcium phosphate (CP) thin films on the physicochemical and biological properties of the films has been investigated in this study. Five different surfaces were used and the culture plate (CTL) and CP film prepared in the absence of magnesium (CaP) were used for the comparison. Three different films were prepared at different magnesium concentrations. CP films prepared at the Mg concentrations of 0.1, 1, and 10 mM were designated as CaPL, CaPM, and CaPH, respectively. The observation of surface morphology of the CP films using scanning electron microscopy (SEM) displayed that the presence of magnesium affected considerably the morphology of the films including a decrease of surface porosity. X-ray diffraction (XRD) analysis for the determination of the crystallinity of the CP films showed that the structure of the films was amorphous. Examination using X-ray photoelectron spectroscopy (XPS) confirmed the prepared films consisted of calcium, phophorus, and magnesium. Cell adhesion assays elucidated that cell adhered less as the magnesium concentration increased. However, cell proliferation assays demonstrated that the cells proliferated more actively on the films prepared at the higher magnesium concentration. These results suggest that the presence of magnesium may promote the biocompatibility of CP films.

• The Korean Journal of Ecology
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• v.17 no.3
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• pp.299-310
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• 1994

Intraspecific clonal interactions have important influences on a population structure of the cellular slime mold (CSM). This study was to investigate whether or not evolutionary change in a population could be induced by clonal competition, and to elucidate how various clones in a population evolve in a homogeneous environment of laboratory culture. The characteristic clones of Polysphondylium pallidum which had different resource consumption rates (RCR) and mating types I and II were selected for study. Investigation was conducted for 4 experimental time interval $(T_0-T_4)$; one experimental time interval took almost 10-14 days from inoculation to havest of fruiting bodies. Two sets of 50 clones were cultured from 50 clones at To, and RCR variations of the population were compared between $(T_0\;and\;T_4)$ for each set of clones. Each clone of the CSM had a diverse resource consumption rate, or growth rate, in a homogeneous and limited Cerophyl agar plate despite the passage of 48-56 generations from the beginning of the experiment. Diverse clones with different growth rate could coexist in one site of the homogeneous agar plate as well as heterogeneous soil microenvironment. When there was high clonal diversity of RCR, a clone in a population had high chances to encounter other clones with resultant increased clonal competition. In one set, 26 of 37 clones of mating type I were changed to mating type Il for the 4 experimental time intervals, which indicated that the rate of competitive exclusion among clones during total experiment from $(T_0\;to\;T_4)$ was 0.703. In another set, 31 of 37 clones of mating type I were changed to mating type II , having the rate of competitive exclusion 0.838. The frequency of each of mat~ng types changed by 0.93-1.29% in each successive generation. The competitive exclusion among clones occurred by 1.26-1.75% when approximately $2.6{\times}10^8$ bacterial cells were provided as food and thereafter one generation of myxamoebae of CSM elapsed at room temperature. This finding implicated that in the vegetative state of P, pallidurn there was 1.26-1.75% probabil~ty of evolutionary change per generation changing from one clone to another clone.

• Clinical and Experimental Pediatrics
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• v.50 no.10
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• pp.1011-1017
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• 2007

Purpose : We tried to assess the optimal conditions to improve low transduction efficiency and their effect on target cells. Methods : Cultured NIH 3T3 cells were incubated with retroviral vectors bearing an enhanced green fluorescent protein (eGFP) gene. We varied the ratio of viral vectors to target cells (1:1-1:8) and the number of transfections (${\times}1$, ${\times}2$), and compared transduction efficiencies. Also, the effects of polybrene on transduction efficiency and viability of target cells were assessed. Transduction of the eGFP gene was evaluated by observing NIH 3T3 cells under a fluorescence microscope and efficiencies were measured by the percentage of eGFP positive cells using FACscan. Results : As the ratio of retroviral vectors to target cells increased, transduction efficiency was greatly improved, from 7% (1:1) to 38% (1:4). However, transduction efficiency did not increase any more when the ratio increased from 1:4 to 1:8. Cells transfected twice showed higher transduction efficiencies than cells transfected once, at a ratio of 1:8. The eGFP gene transduced to NIH 3T3 cells sustained its expression during repeated passages. However, after the third passage (day 9), the percentage of eGFP positive cells began to decline. The degree of this decline in eGFP expression was lower in cells transfected twice than in cells transfected once (P<0.05). The addition of polybrene did not have any toxic effect on NIH 3T3 cells and greatly increased transduction efficiency (P=0.007). In addition to vector component, transduction efficiency was very sensitive to culture confluence. Cells cultured and transfected in 24-well plate showed higher transduction efficiency, although cells cultured in 6- well plate proliferated more (P=0.024). Conclusion : Our data could be used as a basis for retrovirus-based gene therapy. Further study will follow using human cells as target cells.

• Journal of Dental Rehabilitation and Applied Science
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• v.40 no.1
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• pp.13-23
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• 2024

Purpose: This aim of this study was to demonstrate growth factors that differentiate human mesenchymal stem cells into chondrocytes and to evaluate cell proliferation enhancement by needle size differences. Materials and Methods: Human mesenchymal stem cells were cultured in chondrogenic medium supplemented with BMP-2, BMP-4, BMP-6, BMP-7, BMP-13, FGF-2, FGF-18, IGF-1, TGF-β1, TGF-β2, TGF-β3 and without growth factors for 14, 21, and 28 days. Then, the expression levels of SOX-5, SOX-6, SOX-9 and FOXO1A were comparatively analyzed. Human mesenchymal stem cells were inoculated into culture dishes using 18, 21, and 26 gauge (G) needles, and cell proliferation was measured after 24, 48, and 72 hours, respectively. Results: In addition to the previously known FGF, IGF-1, and TGFβ1,the BMP family growth factors such as BMP-2, BMP-4, BMP-6, and BMP-7 increased the expression of chondrocyte differentiation genes SOX-5, SOX-6, SOX-9, and FOXO1A. At 48 hours, the 26G group, the smallest needle, showed significant cell proliferation improvement compared to the control group and the 18G group. At 72 hours, the 26G group, the smallest needle, showed significant increase in cell proliferation compared to the control group. Conclusion: Through this study, growth factors with the ability to induce chondrocyte differentiation of human mesenchymal stem cells were investigated, and cell proliferation changes by needle size differences were determined.