• Archives of Pharmacal Research
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• v.21 no.4
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• pp.398-405
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• 1998

We investigated the effects of triterpene acids (TAs), ursolic acid (UA) and oleanolic acid (OA), on the induction of proliferation and differentiation of normal rat mammary epithelial cells (RMEC) or organoids cultured in Matrigel or primary culture system. To elucidate the effects, we tested their differentiation inducing activities with intercellular communication ability, cell cycle patterns, induction of apoptosis, and morphological differentiation in the three dimensional extracellular culture system. To study the changes of RMEC subpopulation in culture, the cultured cells were isolated, immunostained with peanut lectin (PNA) and anti-Thy-1.1 antibody and then analyzed with flow cytometry. Four different subpopulations, such as PNA and Thy-1.1 negative cells (B-), PNA positive cells (PNA+), Thy-1.1 positive cells (Thy-1.1+), PNA and Thy-1.1 positive cells (B+), were obtained and the size of each subpopulation was changed in culture with time in the presence of TAs. Intercellular communication was observed in culture for 7 days in TAs-treated cells, but not in culture for 4 days with scrape-loading dye transfer technique. $G_2$/M phase cells and the number of apoptotic population were increased in TAs-treated groups in cell cycle analyses. S phase fractions were reduced and the change of $G_1$ phase cells was not observed. The colonies with distinct multicelfular structures, such as stellate, ductal, webbed, squamous, lobulo-ductal colonies, were observed in Matrigel culture and the frequencies of each colony were changed in the presence of TAs. These results suggest that UA and OA have differentiation inducing effects on rat mammary epithelial cells in primary or in Matrigel culture.

• Biomolecules & Therapeutics
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• v.11 no.4
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• pp.224-231
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• 2003

The aim of this study was to investigate the effect of resveratrol on the oxidative stress-induced inhibition of gap junctional intercellular communication in HaCaT keratinocytes. Anti-oxidative activity of resveratrol was measured by $\alpha,\alpha$-diphenyl-$\beta$-picrylhydrazyl assay and dichlorodihydrofluorescein diacetate oxidation assay. Gap junctional intercellular communication in HaCaT keratinocytes was assessed using the scrape loading/dye transfer technique. Western blots and reverse transcription-polymerase chain reaction were also analyzed for connexin 43 protein and mRNA expression, respectively. Resveratrol scavenged directly the stable $\alpha,\alpha$-diphenyl-$\beta$-picrylhydrazyl radical over a concentration range of 4 mg/ml ($78.2{\pm}2.7$% of control) to 500 mg/ml ($29.9{\pm}4.2$% of control) and decreased the intracellular reactive oxygen species induced by ultraviolet A (UVA) irradiation ($89.3{\pm}1.1$% of UVA group), ultraviolet B (UVB) irradiation ($70.9{\pm}1.7$% of UVB group) and 12-0-tet-radecanoylphorbol-13-acetate (TPA, $48.3{\pm}1.1$% of TPA group), respectively. UVA irradiation and TPA markedly reduced gap junctional intercellular communication, which was restored by resveratrol. There were no significant differences in the level of connexin 43 protein and mRNA expression among any of the experimental groups. Our data suggests that resveratrol has the protective effect on the oxidative stress-induced inhibition of gap junctional intercellular communication in HaCaT keratinocytes, and this protection is likely due to the scavenging of reactive oxygen species.

• Proceedings of the Korean Society of Near Infrared Spectroscopy Conference
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• 2001.06a
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• pp.4105-4105
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• 2001

Near infrared spectroscopy (NIRS) was employed to qualify and quantify on survival, the injury rate and apoptosis of the human breast cancer cell line MCF-7 cells. MCF-7 cells were cultured in RPMI medium supplemented with 10% FCS in a 95% air and 5% CO2 atmosphere at 37$^{\circ}C$. For the viable cells preparation, cells were de-touched by 0.1% of trypsin treatment and washed with RPMI supplemented with 10% FCS medium by centrifugation at 1000 rpm for 3min. For the dead cells preparation, cells were de-touched by a cell scraper. The cells were counted by a hemacytometer, and the viability was estimated by the exclusion method with frypan blue dye. Each viable and dead cells were suspended in PBS (phosphate bufferred saline) or milk at the cell density desired. For the quantitative determination of cell death by measuring the LDH (lactate dehydrogenase) activity liberated from cells with cell membrane injuries, LDH-Cytotoxic Test Wako (Wako, Pure Pharmaceutical Co. Ltd., Japan) was used. We found that NIRS measurement of MCF-7 cells at the density range could evaluate and monitor the different characteristics of living cells and dead cells. The spectral analysis was performed in two wavelength ranges and with 1,4, 10 mm pathlength. Different spectral data pretreatment and chemometrics methods were used. We applied SIMCA classificator on spectral data of living and dead cells and obtained good accuracy when identifying each class. Bigger variation in the spectra of living cells with different concentrations was observed when compared to the same concentrations of dead cells. PLS was used to measure the number of cells in PBS. The best model for measurement of dead cells, as well as living cells, was developed when raw spectra in the 600-1098 nm region and 4 mm pathlength were used. Smoothing and second derivative spectral data pretreatment gave worst results. The analysis of PLS loading explained this result with the scatter effect found in the raw spectra and increased with the number of cells. Calibration for cell count in the 1100-2500 nm region showed to be very inaccurate.

• Journal of Biomedical Engineering Research
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• v.38 no.1
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• pp.43-48
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• 2017

Chemotherapy, radiation therapy, and surgery are major methods to treat cancer. However, current cancer treatments report severe side effects and high recurrences. Recent studies about engineering nanoparticles as a drug carrier suggest possibilities in terms of specific targeting and spatiotemporal release of drugs. While many nanoparticles demonstrate lower toxicity and better targeting results than free drugs, they still need to improve their performance dramatically in terms of targeting accuracy, immune responses, and non-specific accumulation at organs. One possible way to overcome the challenges is to make precisely controlled nanoparticles with respect to size, shape, surface properties, and mechanical stiffness. Here, we demonstrate $500{\times}200nm$ discoidal polymeric nanoconstructs (DPNs) as a drug delivery carrier. DPNs were prepared by using a top-down fabrication method that we previously reported to control shape as well as size. Moreover, DPNs have multiple payloads, poly lactic-co-glycolic acid (PLGA), polyethylene glycol (PEG), lipid-Rhodamine B dye (RhB) and Salinomycin. In this study, we demonstrated a potential of DPNs as a drug carrier to treat cancer.

• Journal of Korean Dental Science
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• v.10 no.1
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• pp.29-34
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• 2017

Purpose: The purpose of this study was to evaluate the microleakage of various types of resin-bonded fixed partial dentures (RBFPDs) after artificial aging. Materials and Methods: Forty models with missing first molar were fabricated using artificial resin teeth and were divided into four groups: Group A, conventional RBFPDs design; Group B, modified RBFPDs design; Group C, assembled 3-piece RBFPDs design; and Group D, assembled 3-piece RBFPDs with different occlusal rest positions. Half of the specimens underwent chewing simulation process (240,000 cycles, 50 N load, 1.7 Hz) and thermocycling (temperatures $5^{\circ}C{\sim}55^{\circ}C$, dwelling time 30 seconds) and the remaining 20 specimens didn't receive any treatment. All the specimens were immersed in 2% methylene blue solution for 24 hours to evaluate microleakage, and were sectioned at the middle part of abutment teeth. To evaluate the microleaskage, a dye penetration was calculated. Result: With artificial aging, cyclic loading and thermocycling, a 3-piece RBPFD and a 2-piece RBPFD using original tooth undercuts have significantly lower microleakge (P<0.05) compared to the conventional design of RBPFD and modified RBPFD. Conclusion: Within the limit of this experiment, the assembled RBFPDs exhibited a smaller microleakage than the conventional RBFPDs, implying that the assembled RBFPDs can be more effective for reducing the dislodgement of the RBFPDs.

• Journal of Ginseng Research
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• v.24 no.4
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• pp.188-195
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• 2000

Using Ginseng saponins (crude saponin and total saponin) and ginsenoside Rbl Rb2, Rc, Rd, Re, Rhl, and Rh2 in this study, we have examined the effects of the compounds on the induction of differentiation in normal rat mammary epithelial cells and 7,12-dimethylbenz[a]anthracene (DMBA)-induced mammary tumor cells in culture. When normal rat mammary organoids were cultured in 100-mm culture plates in the presence or absence of ginseng saponins, there were four different cell colonies after two weeks in culture: cobble stone, spindle, honey comb, and senescence type colonies. Ginseng saponins showed different effects on the development of each colonies. Scrape-loading dye transfer tech-nique was performed to measure the effects of total saponin, Rhl, and Rh2 on intercellular junctional communication. Intercellular communication was not observed at short cultilral time, e.g., four or seven days, but when it cultured it up to two weeks, cell to cell communication was observed in saponin-treated cells. Reconstituted basement membrane, Matrigel, supported the growth and development several different multicellular structures from normal mammary organoids (e.g., ductal, webbed, stellate, and squamous colonies) or DMBA-induced mammary tumor (e.g., alveolar unit, foamy alveolar unit, squamous metaplasia, lobule-ductal, stellate, and webbed colony). In ginseng saponin-treated groups, webbed colonies were more and squamous colonies were less than control group. Moreover, the ductal colonies, marker tructure of well-differentiate mammary epithelial cells, were developed more in saponin-treated group than in control group. In conclusion, ginseng saponins affected on the differentiation of normal rat mammary epithelial cells and DMBA-induced mammary tumor cells in culture.

• Journal of Food Hygiene and Safety
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• v.16 no.3
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• pp.212-220
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• 2001

To investigate the modifying effect of Kwao Kreu, Pueraria mirifica (PM), we performed two kind of studies which are the non-surgical medium-term carcinogenicity study and the modulation of gap junctional intercellular communication study. The first study, a non-surgical medium-term carcinogenicity bioassay was done to investigate the modifying effect of Kwao Keru, Pueyaria mirifca (PH), a rejuvenating folk medicine from Thailand, on the male F344 rat liver. Specific pathogen free, male 6-week-old F3444 rats were divided into ten groups. To induce hepatocarcinogenesis, those in all groups were given a single i.p. injection of DEN (200 mg/kg) and were received two i.p. injection of DGA (300 mg/kg) at the ends of weeks 2 and 5. Rats of group 3-6 were given sodium phenobarbital (PB 0.05% in drink). A diet containing 10 mg/kg PM was given to group 2 during the post-initiation phase and to groups 4 and 5 during promotion and initiation phase, respectively. Group 6 was given the experimental diet alone throughout the experiment (8 weeks). Rats of group 7, 8, 9 and 10 were fed 1000 mg/kg PH in the same manner as group 2, 4, 5 and 6. All animals were sacrificed at 8 weeks after DEN administration. Result of the immunohistochemical staining of the glutathione S-transferase placental form (GST-p) indicated that the numbers and areas of the preneoplastic leisions were not significantly changed in all PM treatment group comparing to control group. Also the numbers and areas of GST-p positive foci among group 7, 8, 9 and 10 were not significantly changed in comparing to control group. To study the effect of PM on the modulation of gap junctional intercellular communication, the present study was performed scrape-loading dye transfer (SL/DT) assay in human keratinocytes. The results showed that PM could not modulate GJIC. These results indicate that Pueraria mirifica may have no carcinogenic effects on experimental hepatocarcinogenesis in rats and gap junctional intercellular communication in human keratinocyte.

• Journal of the korean academy of Pediatric Dentistry
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• v.34 no.3
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• pp.359-369
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• 2007

This in vitro study was performed to assess the effect of surface sealing on the microleakage of class V composite resin restorations that underwent several aging treatments. Class V cavities were prepared on the buccal surface of 100 sound extracted premolars and restored with a hybrid light-cured composite resin according to the manufacturer's instructions. They were randomly divided into two groups consisting of 50 samples: group I, without surface sealing, and group II, in which margins were etched and surface sealant was applied. After thermocycling, each group was divided into five subgroups, respectively, to represent the five aging treatments: group A = no further treatment (only thermocycling), B = toothbrushing, C = load cycling, D = toothbrushing followed by load cycling, and E = aging treatment in deionized water for six months. Microleakage was assessed by examining the penetration of 2% methylene blue dye. The following results were obtained: 1. At occlusal and cervical margins in groups without surface sealing, there was no significant difference in microleakage after the several aging treatments (p>0.05). 2. The occlusal margins of groups with surface sealing showed no significant differences after the several aging treatments (p>0.05). 3. In the cervical margins of groups with surface sealing, microleakage significantly increased after load cycling or aging in deionized water for six months (p<0.05). 4. The no-further-treatment group and the toothbrushing group with surface sealing showed less microleakage than the corresponding groups without surface sealing (p<0.05). 5. The surface-sealed groups with load cycling or aging in deionized water showed no significant difference in microleakage to the corresponding groups without surface sealing (p>0.05). In conclusion, the results of this study suggest that the surface sealant infiltrating through the gap of the cervical margin exerted a positive effect on microleakage at the initial stage, but the effect was not sufficient to overcome the stress generated by the cuspal flexure during occlusal loading and water absorption.