Journal of the korean academy of Pediatric Dentistry
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v.24
no.1
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pp.204-219
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1997
The use of fluoride is one of the most effective methods for caries prevention. Fluoridation of public water supply has been recognized, for many years, as an effective way to reduce dental caries. The fluoride supplement has been recommended when the natural fluoride was unavailable or below the optimal range. However the mechanism of caries prevention by fluoride has not yet been clarified and it is well known that an overdose of fluoride results inacute and chronic toxicity, especially dental fluorosis. Fluoride mouthrinsing solution is widely used in dentistry due to its effectiveness in carrying anticariogenic action. Understanding the effects of fluoride mouthrinsing solution on human gingival fibroblasts will provide the safety rationale for its use during the caries preventive therapy. The purpose of this study was to evaluate the cytotoxic effect of fluoride mouthrinsing solution on the human gingival fibroblast in vitro. The human gingival fibroblasts were cultured from healthy gingiva on the extracted deciduous teeth of children. Cells were inoculated into a 24-well plate with $1{\times}10^4cells/well$ of medium at $37^{\circ}C$, 100% humidity, 5% $CO_2$ incubator for 24 hours. And the cells were counted by using the hemocytometer at each designed study. Human gingival fibroblasts were cultured in growth medium after one minute application range of 0.02%-0.2% NaF solution and 0.1% $SnF_2$ solution. The cells used in this study were between fifth to eighth passage number. The cell morphology was examined by inverted microscope and cell proliferation was measured by incorporating $[^3H]$-thymidine into DNA. DNA synthesis by human gingival fibroblasts was assessed by $[^3H]$-thymidine uptake assays while the cell activity was measured by MTT assay. Each concentrated fluoride mouthrinsing solution was estimated for its biocompatability with fibroblasts by the tissue culture technique. The results of this study were as follows : 1. It was observed that at 0.05%, 0.2% NaF mouthrinsing solution the cytoplasmic processes became globular. When 0.1% $SnF_2$ mouthrinsing solution was applied, the cytoplasmic process and cell morphology were disappeared. 2. DNA synthetic activity was reduced regardless of the concentration of the fluoride mouthrinsing solution. However, the result is statistically insignificant except 0.1% $SnF_2$ mouthrinsing solution(p<0.05). 3. Our results indicate that 0.02%, 0.05% concentrations of NaF mouthrinsing solution caused minimal cytotoxicity. But 0.2% NaF and 0.1% $SnF_2$ concentration were a significant difference between the cell activity in the experimental group and control group (p<0.05). 4. After appling 0.05% & 0.02% NaF fluoride mouthrinsing solution, cell activity was restored to the control groups level according to incubating time. The results suggest that direct exposure to fluoride solution inhibits gingival fibroblast activity. Therefore, for the most effective use of fluoride use, lowering the concentration of fluoride mouthrinsing is advisable because it maintains biocompatability and free ion in the oral fluid.
large scale production of cloned embryos requires the technology of multiple generation nuclear transplantation(NT) using NT embryos as the subsequent donor nuclei. The purposes of this study were producing the second generation cloned rabbit embryos, and also to determine the electrofusion rate and in vitro developmental potential comparatively in the cloned embryos of the first and second NT generation. The embryos of 16-cell stage were collected from the mated does by flushing oviducts with Dulbecco's phosphate buffered saline(D-PBS) containing 10% fetal calf serum(FCS) at 47 hours after hCG injection In the first generation NT, the nuclear donor embryos were synchronized in the phase of Gi /S transition of 32-cell stage. The first generation NT embryos which were developed to 8-cell were synchronized in Gi /S transition phase of the following 16-cell stage and used as donor nuclei for second generation Synchronization of the cell cycle of blastomeres was induced, first, using an inhibitor of microtuble polymerization, colcemid for 10 hours to arrest blastomeres in M phase, and secondly, using a DNA synthesis inhibitor, aphidicolin for 1.5 to 2 hours to arrest them in Gi /S transition boundary. The recipient cytoplasms were obtained by removing the nucleus and the first polar body from the oocytes collected at 14 hours after hCG injection. The separated donor blastomeres were injected into the enucleated recipient oocytes by micromanipulation and were electrofused by electrical stimulation of three pulses for 60 $\mu$sec at 1.25 kV /cm in 0.28 M rnannitol solution The fused oocytes were co-cultured with a monolayer of rabbit oviductal epithelial cells in M-199 solution containing 10% FCS for 120 hours at 39$^{\circ}C$ in a 5% $CO_2$ incubator. Following in vitro culture of the first and second generation cloned embryos to blastocyst stage, they were stained with Hoechst 33342 dye for counting the number of blastomeres by fluorescence microscopy. The results obtained were summarized as follows: 1. The electrofusion rate was found to be similar as 79.4 and 91.5% in the first and second generation NT rabbit embryos, respectively. 2. The in vitro developmental potential to blastocyst stage of the second generation NT embryos (23.3%) was found significantly(p<0.05) lower, compared with that of the first generation NT embryos (56.8%). 3. The mean blastomeres counts of embryos developed to blastosyst stage following in vitro culture for 120 hours and also their daily cell cycles during the culture period were decreased significantly (p<0.05) to 104.3 cells and 1.33 cylces in the second NT generation, compoared with 210.4 cells and 1.54 cycles in the first NT generation, respectively.
This study was designed to investigate effect of cryoprotectant kinds and cell stages on the viability of bovine embryos cryopreserved by vitrification. The oocytes were collected from ovarian follicles of Korean native cows. The follicular oocytes were cultured in TCM-199 medium containing hormone and 10%(v/v) FCS for 24~48hrs in a incubator with 5% $CO_2$, in air at 38.5$^{\circ}C$ and then matured oocytes were again cultured for 7~10 hrs with spermatozoa capacitated by preincubation. The vitrification solutions of EFS and EDS were consisted of 40%(v/v) ethylene glycol, 18%(v/v) Ficoll and 0.3M sucrose, and 20%(v/v) ethylene glycol, 16.5%(v/v) DMSO and 0.5M sucrose in TCM-199 medium supplemented with 10% FCS, respectively. The embryos were exposed to EFS or EDS at $25^{\circ}C$ and loaded into OPP straw for 30 sec. The plug end of each straws was heat-sealed and straws was slowly immersed into liquid nitrogen(L$N_2$). The results obtained were summarized as follows : 1 . The rates of cleavage and hatching of embryos frozen with vitrification, rapid and slow freezing methods were 67.5%, 27.5% and 42.5%, 20.0% and 52.5%, 25.0%, respectively And rates of cleavage and hatching of embryos frozen with vitrification method were significantly(p<0.05) higher than those in other methods, and the rates were lower than those in control group(82.5% and 37.5%). 2. The rates of cleavage and hatching of embryos were significantly(p<0.05) different between EFS(47.5% and 22.5%) and EPS(52.5% and 27.5%), and the rates were lower than those in control group(82.5% and 37.5%). 3. After vitrification freezing of bovine embryos at zygote, 2 cell, 8 cell, morulae and blastocyst stage, the rate of cleavage and hatching were 25.0% and 15.0%, 32.5% and 20.0%, 37.5% and 20.0%, 52.5%, 27.5%, 47.5% and 25.0%, respectively. And developmental rates to the expended blastocyst stage of embryos frozen at zygote stage was significantly(p<0.05) lower rather than those in 2, 8-cell and morulae stage.
In the present study, mouse follicular oocytes and 2-cell embryos(late -zygote stage embryos included) were cultured on the electric pad for electric stimulation in the culture incubator. In addition, follicular oocytes and embryos were tested for maturation and development under higher temperature condition($39^{\circ}C$).Mouse follicular oocyte maturation were not affected by anion electric stimulation and there is no significant difference in GBVD and MI between the control and experiment group after 4hr culture. In the embryo culture, it was found that more morula and blastocyst were found in the electric stimulation group rather than the control(96hr). This may seem to be caused with cytoplasmic $Ca^{2+}$ transient rise by electric stimulation(anion pad). On the other hand higher temperature incubation ($39^{\circ}C$) on the anion pad caused all the embryos degenerated within $12h{\sim}24hr$ culture. This was quite different from large animal embryos(bovine, pig, sheep), in which beneficial effect of high temperature incubation for oocyte maturation and embryo development were found.
Healing of periodontal tissues require the migration and proliferation of gingival fibroblasts and periodontal ligament cells. There is many evidences that the some agents like cytokines and polypeptide growth factors are mediate these cellular events in wound healing. Recently someone is interested in herbal drugs on periodontal tissue healing processes. The purpose of this study was to examine the effects of 4 herbal drugs, Carthami Flis, Moutan Redias Cortex, Scirpi Rhisoma, Seed of Carthamus tinctorius L. on human gingival fibroblasts and periodontal ligament cells. Periodontal ligament cells and gingival fibroblasts were primarily cultured from extracted premolar with non-periodontal diseases. The powder from extracted. herbal drugs were prepared with distilled water. Cells were cultured with DMEM at $37^{\circ}C$, 5% $CO_2$, 100% humidity incubator, and treated with each herbal drugs with proper concentration for 1, 2, and 3 days. The cell activity was determined by ELISA reader using MTT assay. There was the most significant elevation in $10^{-3}g/ml$ of almost herbal drugs on cellular activities. The result of this study demonstrated that Carthami Flis, Moutan Radicis Cortex, Scirpi Rhisoma, Seed of Carthamus tinctorius L. appears to have beneficial effect on healing process after periodontal treatment.
This study was carried out to investigate the effects of gonadotropins added during maturation of porcine oocytes on the in vitro maturation(IVM), in vitro fertilization(IVF) and developmental potential of embryos. The follicular oocytes were cultured in TCM-199 medium containing different combination of gonadotropins(5$\mu$g /ml FSR or 1OIU /ml PMSG and 1O$\mu$g /ml LH or 1OIU /ml hCG), 10% FCS and 10% PFF for 36~48h in a incubator with 5% $CO_2$ in Air at 39$^{\circ}C$ and then matured oocytes were again cultured to 120h after IVF for 6~7h with heparin(100$\mu$g /m')-treated sperm. When the oocytes were matured for 42brs in the medium containing FSH+LH, FSH+hCG, PMSG+LH or PMSG+hCG, the JVF rate of each treatment was 50.0%, 52.9%, 66.7% and 70.0%, respectively. The highest CEI (cumulus cell expansion index) was obtained from PMSG+hCG-added medium and the highest polyspermic penetration resulted from FSH+LH-added medium. The cleavage of IVF oocytes derived from hormone added IVM was significantly(P<0.05) promoted by PMSG+hCG and the cleavage rate after 36-h, 42-h and 48-h maturation aws 53.0%, 56.7% and 45.6%, respectively. The highest developmental potential resulted from the oocytes derived from PMSG+LH -added IVM.
Lee, Young Seung;Dzagbletey, Philip Ayiku;Chung, Jae-Young;Jeon, Sang Bong;Lee, Ae-Kyoung;Kim, Nam;Song, Seong Jong;Choi, Hyung-Do
ETRI Journal
/
v.42
no.6
/
pp.837-845
/
2020
The objective of this study was to implement an in vitro exposure system for 28 GHz to investigate the biological effects of fifth-generation (5G) communication. A signal source of 28 GHz for 5G millimeter-wave (MMW) deployment was developed, followed by a variable attenuator for antenna input power control. A power amplifier was also customized to ensure a maximum output power of 10 W for high-power 28-GHz exposure. A 3-dB uniformity over the 80 mm × 80 mm area that corresponds to four Petri dishes of three-dimensional cell cultures can be obtained using a customized choke-ring-type antenna. An infrared camera is employed for temperature regulation during exposure by adjusting the airflow cooling rate via real-time feedback to the incubator. The reported measurement results confirm that the input power control, uniformity, and temperature regulation for 28-GHz exposure were successfully accomplished, indicating the possibility of a wide application of the implemented in vitro exposure system in the fields of various MMW dose-response studies.
The current study was undertaken to determine the effects of sex steroid hormones (estrogen, testosterone and 19-nortestosterone) on proliferation and differentiation of preadipocytes of female and male pigs. The preadipocytes were isolated from the backfat of new-born female and male pigs by collagenase digestion and cultured in the $CO_2$ incubator. The concentration of $10^{-7}M$ and 10-6M sex steroid hormones were treated to the cultured preadipocytes. Regarding the effects on preadipocytes proliferation, high concentration ($10^{-6}M$) of all the three hormones increased proliferation of female preadipocytes,and only estrogen and testosterone increased proliferation of male preadipocytes. Regarding the effects on preadipocyte differentiation, all the three hormones increased differentiation of pig preadipocytes, regardless of hormone concentrations and sex of preadipocytes. The degree of stimulation of cell differentiation by sex steroid hormones was greater than that of cell proliferation.
Journal of the Korea Academia-Industrial cooperation Society
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v.13
no.9
/
pp.4341-4348
/
2012
This paper presents a web-based real-time monitoring system for a photobioreactor using an WiFi wireless network. An WiFi interface can support high speed data transfer, up to 11Mbps and it can be compatible with commercial wireless LAN environment. Thus, the proposed cell culture based on WiFi network can be easily applied to the reconfigurable system and real-time monitoring system. In this paper, we integrate the commercial WiFi module to the various bio-sensors and sensor control board to configure the wireless network. After we evaluate application S/W for monitoring the environment within incubator, we verify the proposed sensor networks for a cell culture system and its monitoring system. This result can be applicable for various bio-applications that require the network configuration and real-time monitoring system.
Journal of the Korea Organic Resources Recycling Association
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v.10
no.3
/
pp.126-131
/
2002
For the probiotic feed production from residual food waste, aerobic liquid fermentation was conducted by thermophilic bacteria. 11 Strains of bacteria were isolated from several soil sources and residual food waste. Screening was carried by shaking incubator for the separation of thermophilic strain at $55^{\circ}C$. The isolated strains were tested for enzyme activities such as ${\alpha}$-amylase and protease. 6 Bacterial strains were chosen and were adapted by repeated fermentation processes in food waste substrate. The viable cell count of them at final fermentation stages were shown as $3-7{\times}10^9/ml$ in 2L-jar fermenter. Among them B3, B6 showed higher enzyme activity. By the mixed fermentation of B3, B6 and Bacillus stearothermophilus, the highest viable cell count reached to $1.4{\times}10^{10}/ml$ in 8 hours.
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