• 치위생과학회지
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• 제19권2호
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• pp.133-140
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• 2019

Background: The light-emitting diode (LED) curing light used is presumed to be safe. However, the scientific basis for this is unclear, and the safety of LED curing light is still controversial. The purpose of this study was to investigate the effect of LED curing light irradiation according to the conditions applied for the polymerization of composite resins in dental clinic on the cell viability and inflammatory response in Raw264.7 macrophages and to confirm the stability of LED curing light. Methods: Cell viability and cell morphology of Raw264.7 macrophages treated with 100 ng/ml of lipopolysaccharide (LPS) or/and LED curing light with a wavelength of 440~490 nm for 20 seconds were confirmed by methylthiazolydiphenyl-tetrazolium bromide assay and microscopic observation. The production of nitric oxide (NO) and prostaglandin $E_2$ ($PGE_2$) was confirmed by NO assay and $PGE_2$ enzyme-linked immunosorbent assay kit. Expression of interleukin $(IL)-1{\beta}$ and tumor necrosis factor $(TNF)-{\alpha}$ in total RNA and protein was confirmed by reverse transcription polymerase chain reaction and Western blot analysis. Results: The LED curing light did not affect the viability and morphology of normal Raw264.7 cells but affected the cell viability and induced cytotoxicity in the inflammation-induced Raw264.7 cells by LPS. The irradiation of the LED curing light did not progress to the inflammatory state in the inflammation-induced Raw264.7 macrophage. However, LED curing light irradiation in normal Raw264.7 cells induced an increase in NO and $PGE_2$ production and mRNA and protein expression of $(IL)-1{\beta}$ and $(TNF)-{\alpha}$, indicating that it is possible to induce the inflammatory state. Conclusion: The irradiation of LED curing light in RAW264.7 macrophage may induce an excessive inflammatory reaction and damage oral tissues. Therefore, it is necessary to limit the long-term irradiation which is inappropriate when applying LED curing light in a dental clinic.

• 동의생리병리학회지
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• 제33권2호
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• pp.131-140
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• 2019

The objective of present study is to evaluate concentration-dependent in vitro anti-osteoarthritic (OA) effects of synergic mixed formula consisted of dried pomegranate juice concentrate powder, Eucommiae Cortex aqueous extract and Achyranthis Radix aqueous extract 5:4:1 (g/g) mixture on the primary cultured rat articular chondrocytes. First, any cytotoxic effect of mixture was observed using MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium Bromide) assay. Next, cyto-protective effect of test substances was evaluated by using the recombinant human interleukin $(rhIL)-1{\alpha}$ induced chondrocytes. In addition, anti-inflammatory effects were also observed on the lipopolysaccaride (LPS) treated chondrocytes through prostaglandin $E_2(PGE_2)$ productions and 5-lipoxygenase (LPO) activities, and inhibitory effects on matrix metalloproteinase (MMP)-2 and MMP-9 activities were observed on $rhIL-1{\alpha}$ treated chondrocytes with their extracellular matrix (ECM) related mRNA expressions. No obvious cytotoxic effects of mixture were demonstrated. Inflammatory damages of chondrocytes and related ECM degradations induced by treatment of LPS or $rhIL-1{\alpha}$ were significantly and concentration-dependently inhibited by pretreatment of mixture from a concentration level of 0.001 mg/ml to 1 mg/ml. In addition, mixture showed $IC_{50}$ for $rhIL-1{\alpha}-induced$ MMP-2 and MMP-9 activities as 44.01 and $162.47{\mu}g/ml$, and also showed $EC_{50}$ for $rhIL-1{\alpha}-induced$ inhibition of collagen type II, SOX9 and aggrecan mRNA expression as 8.61, 10.79 and $4.47{\mu}g/ml$, respectively. It is observed that mixture showed concentration-dependent anti-inflammatory and cytoprotective ECM preserved effects on the primary cultured rat articular chondrocytes without cytotoxicity.

• 한국환경과학회지
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• 제28권2호
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• pp.225-233
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• 2019

In this study, we investigated the in vitro anti-biofilm activities of plant extracts of chives (Allium tuberosum), garlic (Allium sativum), and radish (Raphanus sativus L.) against environment harmful bacteria (gram-positive Staphylococcus aureus and, gram-negative Salmonella typhimurium and Escherichia coli O157:H7). In the paper disc assay, garlic extracts exhibited the highest anti-biofilm activity. The Minimal Inhibitory Concentration (MIC) of all plant extracts was generally higher for gram-negative bacteria than it was for gram-positive bacteria. Gram-negative bacteria were more resistant to plant extracts. The tetrazolium dye (XTT) assay revealed that, each plant extract exhibited a different anti-biofilm activity at the MIC value depending on the pathogen involved. Among the plant extracts tested, garlic extracts (fresh juice and powder) effectively reduced the metabolic activity of the cells of food-poisoning bacteria in biofilms. These anti-biofilm activities were consistent with the results obtained through light microscopic observation. Though the garlic extract reduced biofilm formation for all pathogens tested, to elucidate whether this reduction was due to antimicrobial effects or anti-biofilm effects, we counted the colony forming units of pathogens in the presence of the garlic extract and a control antimicrobial drug. The garlic extract inhibited the E. coli O157:H7 biofilm effectively compared to the control antimicrobial drug ciprofloxacin; however, it did not inhibit S. aureus biofilm significantly compared to ciprofloxacin. In conclusion, garlic extracts could be used as natural food preservatives to prevent the growth of foodborne pathogens and elongater the shelf life of processed foods.

• 한국미생물·생명공학회지
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• 제47권3호
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• pp.465-472
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• 2019

Candida albicans는 기회감염을 유발하는 주요한 병원성 진균 중의 하나이다. 캔디다증 치료과정에서 항진균제에 대한 내성이 흔히 발견되는데, 그 이유는 Candida가 바이오필름을 형성할 수 있기 때문이다. 이전의 연구에서 우리는 단삼(Salvia miltiorriza)의 에탄올추출물이 세포막의 투과성을 변화시키고 세포벽 합성을 저해하여 항캔디다 활성을 나타냄을 밝혔다. 본 연구에서는 10개 C. albicans 임상균주가 형성한 초기단계의 바이오필름을 대상으로 XTT 환원분석법으로 대사활성을 측정하니, $78{\mu}g/ml$ 단삼 에탄올추출물에 의해 바이오필름의 대사활성이 평균 51.3% 감소되었다. C. albicans 세포들이 폴리스티렌 표면에 부착하거나 germ tube를 형성하는 과정에서의 단삼 에탄올추출물의 영향을 현미경으로 분석하니, $39{\mu}g/ml$ 단삼 에탄올추출물에 의해 부착된 세포의 밀도는 현저하게 감소하였으나 germ tube 형성은 거의 억제하지 못했다. 단삼 에탄올추출물이 C. albicans SC5314 세포의 균사에 특이적인 유전자 발현에 미치는 영향을 qPCR로 분석한 결과, EAP1은 34.7% (p < 0.001), ALS1은 45.0% (p < 0.001), ALS3는 48.1% (p < 0.001), ECE1은 21.3% (p = 0.006) 억제하였다. 결론적으로 단삼의 에탄올추출물은 초기단계의 C. albicans 바이오필름 발달을 효율적으로 저해하며, 이는 EAP1, ALS1, ALS3 유전자의 발현억제에 따른 세포부착 억제와 관련이 있다. 더불어 단삼 에탄올추출물의 C. albicans 세포막 기능저해와 세포벽 합성억제에 의한 구조변화 또한 세포부착단계에서의 바이오필름 발달억제에 기여할 것으로 추정된다.

• 대한소아치과학회지
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• 제46권1호
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• pp.85-92
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• 2019

이 연구의 목적은 유치 줄기세포에 대해 다양한 Calcium silicate-based material (CSM)의 세포독성을 비교하고 평가하는 것이다. Retro $MTA^{(R)}$ (RM), $EZ-Seal^{TM}$ (EZ), ENDOCEM $Zr^{(R)}$ (EN)의 powder를 세포 배지를 이용하여 용출시키고 filtering 하였다. 유치 줄기세포가 다양한 농도의 용출액 하 배양되었다. MTS assay를 통해 CSM 용출액이 세포 증식에 미치는 영향을 분석하였고, Flow cytometry analysis를 통해 세포 표현형의 변화 여부를 관찰하였다. 10% 농도의 용출액에서 배양된 유치 줄기세포의 흡광도 값은 RM > EN > EZ 순이었다(p = 0.0439). 그러나 유치 줄기세포는 재료에 관계없이 간엽줄기세포의 표현형을 유지했다. 세 종류의 CSM이 줄기세포 marker를 변형시키진 않았지만, EZ가 RM과 EN보다 세포적합성이 낮음을 알 수 있었다.

• Journal of Nutrition and Health
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• 제54권5호
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• pp.448-458
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• 2021

본 연구는 뽕나무 열매 착즙 분말 (MJ)이 지방세포 분화과정에서 염증에 관여하는 유전자 발현 및 miR-132/143 조절에 미치는 효과에 중점을 두고 조사하였다. MJ는 3T3-L1 지방세포 분화 동안 지질축적이 억제되었고 PPAR-γ, CEBP-α 및 aP2와 같은 지방형성 유전자 발현을 억제하였다. 또한 염증 조절 매개체인 TNF-α, IL-6, MCP-1 및 iNOS 유전자 발현이 MJ 처리에 의해 감소되었다. MJ의 지질 축적 억제 효과는 염증과 지방분화에 관여하는 miR-132/143의 발현 감소와 관련이 있음을 확인하였다. 따라서 MJ는 염증을 동반하는 비만 예방 및 치료에 도움이 될 수 있는 식품 소재로서 가능성이 있음을 시사하고 있다. 그러나 MJ의 주요 성분인 안토시아닌의 생체이용율은 1-2% 미만으로 추정되며 예상되는 표적 조직이나 혈류에서 미량 검출되는 것으로 알려져 있다 [33]. 이를 해결하기 위해서는 생체 내 동물실험을 통한 다각적인 분석이 향후 진행되어야 할 것으로 판단된다.

• Journal of Nutrition and Health
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• 제53권6호
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• pp.583-595
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• 2020

본 연구에서는 괭생이 모자반 추출물의 소포체 스트레스 억제 효능을 연구하기 위하여 HepG2 간세포에 PA를 처리하여 소포체 스트레스를 유발한 후 추출물을 처리하여 UPR 관련 인자 발현 정도를 측정하였다. PA 750 μM 처리 시 UPR 관련 인자 (p-IRE1α, p-eIF2α, CHOP)의 단백질 발현이 가장 높게 나타나 소포체 스트레스를 효과적으로 유도함을 확인하였고 PA 750 μM를 12시간 처리 시 UPR 관련 인자 (p-IRE1α, p-eIF2α, CHOP)의 단백질 발현이 가장 높음을 확인하였다. 괭생이 모자반 처리 시 PA에 의해 상향 조절된 UPR 관련 인자의 mRNA 및 단백질 발현이 감소하여 PA로 유도된 소포체 스트레스에 대한 억제 효능이 있음을 보여주었다. 또한, 괭생이 모자반은 SIRT2, SIRT6 및 SIRT7의 mRNA의 발현을 증가시킴으로써 괭생이 모자반의 소포체 스트레스 억제 효능이 SIRT에 의한 것으로 확인되었다. 이러한 결과는 괭생이 모자반이 다양한 소포체 스트레스 관련 질병의 예방과 치료에 활용가능성이 있음을 시사한다.

• Fisheries and Aquatic Sciences
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• 제24권3호
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• pp.109-120
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• 2021

Protein hydrolysates and krill meal (KM) are used as protein sources in aquafeeds. The study was conducted to examine the supplemental effects of shrimp protein hydrolysates (SH) or KM in a high-plant-protein diet for red seabream (Pagrus major). A fish meal (FM)-based diet (40%) was considered as the high-FM diet (HFM) and a diet containing 25% FM and soy protein concentrate, in the expense of FM protein from HFM diet, was considered as the low fish meal (LFM) diet. Two other experimental diets (SH and KM) were prepared by including SH and KM into LFM diet at 5% inclusion levels in exchange of 5% FM from the LFM diet. A feeding trial was conducted for fifteen weeks using triplicate group of fish (Initial mean body weight, 8.47 ± 0.05 g) for a diet. Growth performance and feed efficiency of fish were significantly enhanced by HFM, KM and SH supplemented diets over those of fish fed LFM diet. Interestingly, these parameters of fish fed SH diet showed better performance than KM and HFM groups. Liver IGF-I expression of fish fed SH diet was comparable to HFM group and higher than KM and LFM diets. Protein digestibility of SH diet was significantly higher than KM, HFM, and LFM diets. Dry matter digestibility of SH diet was comparable to HFM diet and significantly higher than KM and LFM diets. Nitro blue tetrazolium and superoxide dismutase activities of HFM, SH and KM groups were significantly elevated than the LFM group and SH diet increased catalase and glutathione peroxidase activities of fish compared to KM and LFM groups. Hemoglobin level and hematocrit of fish fed SH and KM diets were significantly higher than LFM group. A diet containing 20% FM with KM is comparable to a HFM diet which contains 40% FM for red seabream. SH can be used to replace FM from red seabream diet down to 20% and fish performance can be improved better than a diet containing 40% FM. Overall, it seems that SH is more effective ingredient in red seabream diet compared to KM.

• Journal of Ginseng Research
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• 제45권2호
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• pp.228-235
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• 2021

Backgroud: Ginsenoside compound K (GK) is a major metabolite of protopanaxadiol-type ginsenosides and has remarkable anticancer activities in vitro and in vivo. This work used an ionic cross-linking method to entrap GK within O-carboxymethyl chitosan (OCMC) nanoparticles (Nps) to form GK-loaded OCMC Nps (GK-OCMC Nps), which enhance the aqueous solubility and stability of GK. Methods: The GK-OCMC Nps were characterized using several physicochemical techniques, including x-ray diffraction, transmission electron microscopy, zeta potential analysis, and particle size analysis via dynamic light scattering. GK was released from GK-OCMC Nps and was conducted using the dialysis bag diffusion method. The effects of GK and GK-OCMC Nps on PC3 cell viability were measured by using the 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide assay. Fluorescent technology based on Cy5.5-labeled probes was used to explore the cellular uptake of GK-OCMC Nps. Results: The GK-OCMC NPs had a suitable particle size and zeta potential; they were spherical with good dispersion. In vitro drug release from GK-OCMC NPs was pH dependent. Moreover, the in vitro cytotoxicity study and cellular uptake assays indicated that the GK-OCMC Nps significantly enhanced the cytotoxicity and cellular uptake of GK toward the PC3 cells. GK-OCMC Nps also significantly promoted the activities of both caspase-3 and caspase-9. Conclusion: GK-OCMC Nps are potential nanocarriers for delivering hydrophobic drugs, thereby enhancing water solubility and permeability and improving the antiproliferative effects of GK.

• Journal of Korean Neurosurgical Society
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• 제65권6호
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• pp.790-800
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• 2022

Objective : EID3 (EP300-interacting inhibitor of differentiation) was identified as a novel member of EID family and plays a pivotal role in colorectal cancer development. However, its role in glioma remained elusive. In current study, we identified EID3 as a novel oncogenic molecule in human glioma and is critical for glioma cell survival, proliferation and invasion. Methods : A total of five patients with glioma were recruited in present study and fresh glioma samples were removed from patients. Four weeks old male non-obese diabetic severe combined immune deficiency (NOD/SCID) mice were used as transplant recipient models. The subcutaneous tumor size was calculated and recorded every week with vernier caliper. EID3 and AMP-activated protein kinase α1 (AMPKα1) expression levels were confirmed by real-time polymerase chain reaction and Western blot assays. Colony formation assays were performed to evaluate cell proliferation. Methyl thiazolyl tetrazolium (MTT) assays were performed for cell viability assessment. Trypan blue staining approach was applied for cell death assessment. Cell Apoptosis DNA ELISA Detection Kit was used for apoptosis assessment. Results : EID3 was preferentially expressed in glioma tissues/cells, while undetectable in astrocytes, neuronal cells, or normal brain tissues. EID3 knocking down significantly hindered glioma cell proliferation and invasion, as well as induced reduction of cell viability, apoptosis and cell death. EID3 knocking down also greatly inhibited tumor growth in SCID mice. Knocking down of AMPKα1 could effectively rescue glioma cells from apoptosis and cell death caused by EID3 absence, indicating that AMPKα1 acted as a key downstream regulator of EID3 and mediated suppression effects caused by EID3 knocking down inhibition. These findings were confirmed in glioma cells generated patient-derived xenograft models. AMPKα1 protein levels were affected by MG132 treatment in glioma, which suggested EID3 might down regulate AMPKα1 through protein degradation. Conclusion : Collectively, our study demonstrated that EID3 promoted glioma cell proliferation and survival by inhibiting AMPKα1 expression. Targeting EID3 might represent a promising strategy for treating glioma.