• Journal of Animal Science and Technology
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• 제56권8호
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• pp.33.1-33.8
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• 2014

Background: Sox 9 is a major marker of chondrocyte differentiation. When chondrocytes are cultured in vitro they progressively de-differentiate and this is associated with a decline in Sox 9 expression. The active form of vitamin D, 1, 25 $(OH)_2D_3$ has been shown to be protective of cartilage in both humans and animals. In this study equine articular chondrocytes were grown in culture and the effects of 1, 25 $(OH)_2D_3$ upon Sox 9 expression examined. The expression of the transient receptor potential vanilloid (TRPV) ion channels 5 and 6 in equine chondrocytes in vitro, we have previously shown, is inversely correlated with de-differentiation. The expression of these channels in response to 1, 25 $(OH)_2D_3$ administration was therefore also examined. Results: The active form of vitamin D (1, 25 $(OH)_2D_3$ when administered to cultured equine chondrocytes at two different concentrations significantly increased the expression of Sox 9 at both. In contrast 1, 25 $(OH)_2D_3$ had no significant effect upon the expression of either TRPV 5 or 6 at either the protein or the mRNA level. Conclusions: The increased expression of Sox 9, in equine articular chondrocytes in vitro, in response to the active form of vitamin D suggests that this compound could be utilized to inhibit the progressive de-differentiation that is normally observed in these cells. It is also supportive of previous studies indicating that $1{\alpha}$, 25-dihydroxyvitamin $D_3$ can have a protective effect upon cartilage in animals in vivo. The previously observed correlation between the degree of differentiation and the expression levels of TRPV 5/6 had suggested that these ion channels may have a direct involvement in, or be modulated by, the differentiation process in vitro. The data in the present study do not support this.

• 한국수정란이식학회지
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• 제19권3호
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• pp.245-255
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• 2004

포유동물 난자의 체외수정은 외래유전자 도입에 의한 형질전환동물 생산과 우수한 형질을 가진 개체의 보존, 인간의 불임연구 등과 같은 수정란이식 기술로서 널리 이용되고 있다. 돼지 난포란을 이용한 체외수정란의 생산은 초기단계인 체외성숙 기술의 미확립, 그로인한 체외수정 시 높은 다정자 침입율과 불완전한 웅성전핵 형성 몇 체외발달능 정지현상(cell blocking) 등 어려움 때문에 아직도 다른 가축보다 양질의 수정란을 생산하기가 어려운 것으로 알려져 있다. 이와 같은 것을 해결하기 위하여 많은 연구자들은 배양액내에 hormon, growth factor, antioxdants 등과 같은 외인성 인자들을 첨가하고 있다. 이들 인자 중 antioxdant는 free radical을 소거하고 과산화물 생성을 억제하여 난자를 산화적 스트레스로부터 보호한다. 따라서 본 연구는 돼지 난포란으로부터 체외수정란의 생산에 있어서 배양액내 cysteine, catalase 및 glutathione의 첨가가 체외성숙, 체외수정 및 체외배양에 어떤 영향을 미치는지를 검토하였다. 실험 1은 체외성숙용 배양액인 TCM-199 용액에 catalase(100, 200, 500U/$m\ell$)와 glutathione(0.5, 1.0, 1.5mM/$m\ell$)의 첨가, 실험 2는 성숙된 난자의 체외수정용 배양액인 mTBM 용액에 cysteine(0.1, 1.6, 1.0mM/$m\ell$), catalase(100, 200, 500U/$m\ell$)와 glutathione(0.5, 1.0, 1.5mM/$m\ell$)의 첨가, 실험 3은 체외성숙 및 체외수정된 난자의 체외배양용 배양액인 NCSU-23 용액에 cysteine(0.1, 1.6, 1.0mM/$m\ell$), catalase(100, 200, 500U/$m\ell$)와 glutathione(0.5, 1.0, 1.5mM/$m\ell$)을 첨가하여 배반포로의 배 발달율을 관찰하였던 결과는 다음과 같다. 1. 체외성숙시 catalase의 경우는 500U 첨가군의 27.2%로 무첨가군의 15.4%보다 유의하게 높았다(p<0.05). 한편 glutathione의 경우 배반 포로의 배 발달율은 무첨가군과의 차이는 없었다. 그러나 1.0mM 첨가군에서 상실배까지의 배 발달율인 72%는 무첨가군의 53.9%보다 유의하게 높았다(p<0.05). 2. 체외수정시 여러 종류의 항산화제 첨가는 첨가하는 농도와 관계없이 배반포로의 배 발달율은 무첨가군과 차이가 없었다. 3. 체외배양시 여러 종류의 항산화제 첨가는 첨가하는 농도와 관계없이 배반포로의 배 발달율은 무첨가군과 차이가 없었다. 이상의 결과를 종합적으로 하면 돼지 난포란을 이용한 배반포의 체외생산에 있어서 배양액내 항산화제의 첨가는 체외성숙단계에서만 효과적이었다. 이것은 아마도 항산화제가 체외성숙 시 난포란 내에서 일어나는 여러 가지 생화학 반응의 처리시간과 관련하여 활성화시킴으로써 난포란의 생존력을 높인 것이라고 사료되기 때문에 앞으로는 돼지 난포란의 효율적인 체외성숙에 대해서 배양액내 첨가물질은 물론 나아가서 방법론적인 측면에서 더욱 연구되어져야 할 것이다.

• 대한화장품학회지
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• 제40권1호
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• pp.1-10
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• 2014

현재, 기능성 화장품 성분의 피부흡수에 대한 연구 보고가 부족하며, 인체에서의 in vivo 피부 흡수는 거의 보고된 바 없다. 본 연구에서는 in vivo 라만 분광 피부 측정기를 이용하여 대표적 기능성 화장품 8종 성분의 인체 표피 투과 데이터를 수집 및 분석하고, 이를 in vitro 결과와 비교하여 주요 성분의 피부 흡수 자료를 확보하였다. 그리고 본 연구에 사용한 성분의 피부 투과도 측정한 결과를 보면, in vitro 평가에서 ascorbyl-2-glucoside, retinol, retinyl palmitate, 그리고 kojic acid가 우수한 피부 투과율을 보였으며, in vivo 평가에서는 retinol, vitamin C, 그리고 arbutin 이 우수한 것으로 나타났다. 반면, In vitro 평가와 in vivo 평가에서 가장 우수한 투과율을 보인 성분은 각각 ascorbyl-2-glucoside와 retinol이었다. 이러한 차이점은 in vitro 평가의 경우 무모생쥐 모델의 피부를 모두 관통하는 물질을 평가하였고, 라만 분광 피부 측정기의 경우는 표피의 각질층을 평가하였으므로 물질의 특성에 따라서 표피 및 진피층의 영향에 민감할 수 있기 때문인 것으로 판단된다. 그리고 대부분 물질의 투과 장벽은 각질층이므로 각질층에서 물질 이동을 살펴보는 것이 의미 있다고 생각된다. 결론적으로, 본 연구에서는 라만 분광 피부 측정기를 이용하여 대표적 기능성 화장품 성분의 피부 흡수에 대한 기초적인 자료를 확보하였으며, 따라서 이들 자료는 추후 많은 연구에 활용될 수 있을 것으로 생각된다.

• Nutrition Research and Practice
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• 제8권5호
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• pp.487-493
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• 2014

BACKGROUND/OBJECTIVES: D. candidum is a traditional Chinese food or medicine widely used in Asia. There has been little research into the anticancer effects of D. candidum, particularly the effects in colon cancer cells. The aim of this study was to investigate the anticancer effects of D. candidum in vitro and in vivo. MATERIALS/METHODS: The in vitro anti-cancer effects on HCT-116 colon cancer cells and in vivo anti-metastatic effects of DCME (Dendrobium canidum methanolic extract) were examined using the experimental methods of MTT assay, DAPI staining, flow cytometry analysis, RT-PCR, and Western blot analysis. RESULTS: At a concentration of 1.0 mg/mL, DCME inhibited the growth of HCT-116 cells by 84%, which was higher than at concentrations of 0.5 and 0.25 mg/mL. Chromatin condensation and formation of apoptotic bodies were observed in cancer cells cultured with DCME as well. In addition, DCME induced significant apoptosis in cancer cells by upregulation of Bax, caspase 9, and caspase 3, and downregulation of Bcl-2. Expression of genes commonly associated with inflammation, NF-${\kappa}B$, iNOS, and COX-2, was significantly downregulated by DCME. DCME also exerted an anti-metastasis effect on cancer cells as demonstrated by decreased expression of MMP genes and increased expression of TIMPs, which was confirmed by the inhibition of induced tumor metastasis in colon 26-M3.1 cells in BALB/c mice. CONCLUSIONS: Our results demonstrated that D. candidum had a potent in vitro anti-cancer effect, induced apoptosis, exhibited anti-inflammatory activities, and exerted in vivo anti-metastatic effects.

• 한국수정란이식학회지
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• 제12권2호
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• pp.161-169
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• 1997

This study was conducted to compare the insemination time of bovine oocytes and determine the effects of glucose(1.5 mM) on the development of bovine embryos at early cleavage stage. Oocytes were matured for 24 h, followed by exposure to sperm and cultured in modified Tyrode's media drops or with bovine oviduct epithelial cell monolayer prepared in TCM199(BOECM). Insemination time and culture system were varied in each experiment. In experiment 1, to investigate the developmental capacity of bovine embryos after different time of exposure to sperm, bovine ova and sperm were co-incubated for 18, 30 or 54 h, respectively. The development to blastocysts of 30 and 54 h insemination groups were significantly higher(P<0.05) than 18 h group, and in case of blastocysts of cleaved embryos, 30 h group were significantly higher(P<0.05) than other groups. In experiment 2, we investigated the effect of glucose on early bovine embryos. After 18 h insemination, in vitro fertilized oocytes were separated following 3 groups ; G+0, C+24 and C+48. Oocytes of G+0 group were cultured in glucose added Tyrode's medium after fertilization, oocytes in C+24 and C+48 groups were cultured in glucose free Tyrode's medium after fertilization. After 24 h culture, G+24 group was moved to glucose added medium. All oocytes of 3 groups were moved to BOECM after 48 h culture. The rates of cleavage and development to blastocysts in G+0 group were significantly lower than other groups. In experiment 3, we determined the effects of glucose exposure from 8 to 20 h after insemination on the cleavage and development of oocytes. The oocytes in glucose added group had high capacity of cleavage and further development. This study shows that in bovine oocytes, the optimal exposure to sperm is 30 h and glucose exposure to bovine one-cell embryos is detrimental to their first cleavage and further development in vitro but there has no evidence of detrimental effect of glucose(1.5 mM) exposure to bovine embryos over the two-cell stage in vitro.

• Nutrition Research and Practice
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• 제8권4호
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• pp.377-385
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• 2014

BACKGROUND/OBJECTIVES: Abundant consumption of seaweeds in the diet is epidemiologically linked to the reduction in risk of developing cancer. In larger cases, however, identification of particular seaweeds that are accountable for these effects is still lacking, hindering the recognition of competent dietary-based chemo preventive approaches. The aim of this research was to establish the antiproliferative potency and angiosuppressive mode of action of Stoechospermum marginatum seaweed methanolic extract using various experimental models. MATERIALS/METHODS: Among the 15 seaweeds screened for antiproliferative activity against Ehrlich ascites tumor (EAT) cell line, Stoechospermum marginatum extract (SME) was found to be the most promising. Therefore, it was further investigated for its anti-proliferative activity in-vitro against choriocarcinoma (BeWo) and non-transformed Human embryonic kidney (HEK 293) cells, and for its anti-migratory/tube formation activity against HUVEC cells in-vitro. Subsequently, the angiosuppressive activity of S. marginatum was established by inhibition of angiogenesis in in-vivo (peritoneal angiogenesis and chorioallantoic membrane assay) and ex-vivo (rat cornea assay) models. RESULTS: Most brown seaweed extracts inhibited the proliferation of EAT cells, while green and red seaweed extracts were much less effective. According to the results, SME selectively inhibited proliferation of BeWo cells in-vitro in a dose-dependent manner, but had a lesser effect on HEK 293 cells. SME also suppressed the migration and tube formation of HUVEC cells in-vitro. In addition, SME was able to suppress VEGF-induced angiogenesis in the chorio allantoic membrane, rat cornea, and tumor induced angiogenesis in the peritoneum of EAT bearing mice. A decrease in the microvessel density count and CD31 antigen staining of treated mice peritoneum provided further evidence of its angiosuppressive activity. CONCLUSIONS: Altogether, the data underline that VEGF mediated angiogenesis is the target for the angiosuppressive action of SME and could potentially be useful in cancer prevention or treatment involving stimulated angiogenesis.

• BMB Reports
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• 제36권4호
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• pp.387-389
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• 2003

The young developing leaves of willow (Salix safsaf, Salicaceae) trees have antileukemic activity. After a 24-h incubation in vitro, the crude water extracts of the leaves killed a majority of the blasts of acute myeloid leukemia (AML, 73.8%).

• 한국수정란이식학회:학술대회논문집
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• 한국수정란이식학회 2004년도 제4회 발생공학 국제심포지움 및 학술대회
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• pp.90-90
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• 2004

• 대한치과기공학회지
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• 제21권1호
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• pp.145-151
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• 1999

Titanium(Ti) alloys has been mostly concerned with biocompatibility, corrosion resistance, and biofunctionality. However, very little is known, about the biological effects of titanium on microorganism and in particular on the oral flora. So, the effect of titanium on the in vitro growth of oral microorganism forming dental caries was studied under either aerobic or anaerobic condition. In this study, the mostly bacterial species commonly found in dental plaque or gingival sulcus grew well in an aqueous medium containing $100{\mu}g/ml$ of titanium standard solution.

• 대한화장품학회:학술대회논문집
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• 대한화장품학회 1992년도 자외선 차단 화장품의 SPF에 관한 심포지움(대한화장품학회)
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• pp.46-68
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• 1992

To investigate the effect on the antimicrobial activity against S.aureus ATCC 6538, E.coli KCTC 1039 and cell toxic level against transformed mouse fibroblast L929 in formula added with various concentrations of UVB blockers commonly used in cosmetic products, these experiments were carried out by preservative efficacy testing methods and in vitro cytotoxicity methods. The results obtained were as follow ; 1) Octyl Dimethyl PABA had a broad antibacterial spectrum against the Gram (+) and the Gram(-) bacteria at 5.84 % concentration, but not Octyl Methoxycinnamate. 2) Antibacterial activity was decreased in a combined UVB blocker system of squalane base. Especially, Octyl Dimethyl PABA was inactivated by Octyl Methoxycinnamate at 5.84% concentration to a large extents , but not 4-Methylbenzylidene Camphor. 3) Within in vitro cytotoxicity by use of mouse fibroblast L929 on UV-B blockers, NR assay was more excellent than MTT assay on quantitative