• 구강생물연구
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• 제42권4호
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• pp.187-197
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• 2018

Chemical agents such as minocycline (MC) and citric acid (CA) were suggested in the treatment of contaminated implant surface. In this study, MC-HCl treatment was performed to enhance surface characteristics of titanium alloy surface. The purpose of this study was to assess the characteristics and the biocompatibility of Ti-6Al-4V surface treated by MC. Alpha-beta titanium alloy (Ti-6Al-4V) samples were prepared and they were divided into 6 groups according to chemical concentration and treatment time. These groups include 1) group I, non-treated smooth titanium alloy; 2) group II, MC 1.5 mg/mL for 1 hour; 3) group III, MC 1.5 mg/mL for 24 hours; 4) group IV, MC 15 mg/mL for 10 minutes; 5) group V, MC 100 mg/mL for 5 minutes; 6) group VI, pH1 CA for 3 minutes. The analysis of the surface characteristics of MC-treated titanium alloy was executed using scanning electron microscopy, roughness test, and X-ray photoelectron spectroscopy (XPS). Cell adhesion and MTT assay was done using MC3T3 cell. Titanium surfaces treated with MC indicated a more smoothened surface microstructure. For group II and III, the new peaks of rutile TiO2 were found. Group II and V have more basic group of Ti-OH form in XPS. In MTT assay, all MC-treated groups showed significantly higher cell viability compared to control. The surface roughness, crystal structure, surface hydrophilicity, cell viability of smooth titanium surface was improved by MC treatment. Compared with the control experiment and CA-treated group, smooth titanium surface treated with MC showed improved surface characteristics and cell biocompatibility.

• Journal of Microbiology
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• 제44권5호
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• pp.548-555
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• 2006

FaeG is the key factor in the infection process of K88ad enterotoxigenic Escherichia coli (ETEC) fimbrial adhesin. In an attempt to determine the possibility of expressing recombinant FaeG with immunogenicity for a new safe and high-production vaccine in E. coli, we constructed the recombinant strain, BL21 (DE3+K88), which harbors an expression vector with a DNA fragment of faeG, without a signal peptide. Results of 15% SDS-polyacrylamide slab gel analysis showed that FaeG can be stably over-expressed in BL21 (DE3+K88) as inclusion bodies without FaeE. Immunoglobulin G (IgG) and M (IgM) responses in pregnant pigs, with boost injections of the purified recombinant FaeG, were detected 4 weeks later in the sera and colostrum. An in vitro villius-adhesion assay verified that the elicited antibodies in the sera of vaccinated pigs were capable of preventing the adhesion of K88ad ETEC to porcine intestinal receptors. The protective effect on the mortality rates of suckling piglets born to vaccinated mothers was also observed one week after oral challenge with the virulent ETEC strain, $C_{83907}$ (K88ad, $CT^+,\;ST^+$). The results of this study proved that the adhesin of proteinaceous bacterial fimbriae or pili could be overexpressed in engineered E. coli strains, with protective immune responses to the pathogen.

• Journal of Periodontal and Implant Science
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• 제41권6호
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• pp.293-301
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• 2011

Purpose: We have previously reported that tetra-cell adhesion molecule (T-CAM) markedly enhanced the differentiation of osteoblast-like cells grown on anorganic bone mineral (ABM). T-CAM comprises recombinant peptides containing the Arg- Gly-Asp (RGD) sequence in the tenth type III domain, Pro-His-Ser-Arg-Asn (PHSRN) sequence in the ninth type III domain of fibronectin (FN), and the Glu-Pro-Asp-Ilu-Met (EPDIM) and Tyr-His (YH) sequence in the fourth fas-1 domain of ${\beta}$ig-h3. Therefore, the purpose of this study was to evaluate the cellular activity of osteoblast-like cells and the new bone formation on ABM coated with T-CAM, while comparing the results with those of synthetic cell binding peptide (PepGen P-15). Methods: To analyze the cell viability, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay was performed, andto analyze gene expression, northernblot was performed. Mineral nodule formations were evaluated using alizarin red stain. The new bone formations of each group were evaluated using histologic observation and histomorphometrc analysis. Results: Expression of alkaline phosphatase mRNA was similar in all groups on days 10 and 20. The highest expression of osteopontin mRNA was observed in the group cultured with ABM/P-15, followed by those with ABM/T-CAM and ABM on days 20 and 30. Little difference was seen in the level of expression of collagen type I mRNA on the ABM, ABM/T-CAM, and ABM/P-15 cultured on day 20. There were similar growth and proliferation patterns for the ABM/T-CAM and ABM/P-15. The halo of red stain consistent with $Ca^{2+}$ deposition was wider and denser around ABM/T-CAM and ABM/P-15 particles than around the ABM particles. The ABM/T-CAM group seemed to have bone forming bioactivity similar to that of ABM/P-15. A complete bony bridge was seen in two thirds of the defects in the ABM/T-CAM and ABM/P-15 groups. Conclusions: ABM/T-CAM, which seemed to have bone forming bioactivity similar to ABM/P-15, was considered to serve as effective tissue-engineered bone graft material.

• 대한한방내과학회지
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• 제41권4호
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• pp.553-562
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• 2020

Objective: To investigate the vasodilatory effect of Sipjeondaebo-tang by inhibiting RhoA activity in vascular cells during cold exposure. Methods: Human vascular endothelial cells and pericytes were pretreated with Sipjeondaebo-tang for 30 min, followed by incubation at 37 ℃ (control) or 25 ℃ (cold exposure) for 30 min. Activation of endothelin-1-mediated RhoA in pericytes was assessed by pretreating the cells with Sipjeondaebo-tang for 30 min, followed by incubation with endothelin-1 at 37 ℃ for 30 min. Western blotting was performed to measure the expression of active RhoA. Endothelin-1 and nitric oxide release from endothelial cells was examined with enzyme-linked immunosorbent assay kits. The formation of stress fibers and focal adhesion complexes was analyzed by immunocytochemistry. Results: Cold treatment activated RhoA in both pericytes and vascular endothelial cells, whereas Sipjeondaebo-tang treatment inhibited this activation. Sipjeondaebo-tang treatment also reversed the cold-mediated production of endothelin-1 and nitric oxide. Cold exposure promoted the formation of stress fibers and focal adhesion complexes by increasing the expression of phospho-focal adhesion complex kinase, whereas Sipjeondaebo-tang treatment suppressed this response. Conclusions: These findings suggested that Sipjeondaebo-tang inhibits cold-induced RhoA activation and its related pathway components, including endothelin-1 and nitric oxide, in vascular cells. Therefore, Sipjeondaebo-tang could be beneficial for the treatment of Raynaud's phenomenon.

• 폴리머
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• 제27권5호
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• pp.397-404
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• 2003

조직공학적 지지체에서의 골수유래 간엽줄기세포의 점착과 성장에 있어서 세포 점착성 단백질과 폴리펩타이드와의 상호작용을 조사하였다. 세포 점착성 물질로 알려진 단백질이나 폴리펩타이드는 락타이드-글리콜라이드 공중합체인 PLGA 필름과 지지체에 흡착하여 코팅되었으며, 이에 골수유래 간엽줄기 세포의 점착과 성장 거동을 비교하였다. 이들 단백질과 폴리펩타이드는 콜라겐 IV형과 피브리노겐, 라미닌, 젤라틴, 피브로넥틴, 폴리(L-라이신)이 사용되었다. 이중 폴리(L-라이신)을 제외한 단백질과 폴리펩타이드는 PLGA 필름 표면에 거의 단층으로 덮어져 흡착되었으며, PLGA 필름과 지지체에서 골수유래 간엽줄기세포가 1일과 2일, 4일간 배양되었다. 세포의 점착과 성장 거동은 sulforhodamine B법으로 평가하였다. PLGA 필름과 지지체에 단백질이나 폴리펩타이드가 흡착되지 않은 표면보다는 흡착된 표면에서의 세포의 점착과 성장이 우수하였다.

• 한국식품영양과학회지
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• 제43권7호
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• pp.1009-1016
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• 2014

묵은지 스타터로 개발된 Lb. curvatus ML17의 생균제적 특성 및 기능성 조사를 통하여 기능성 소재로서의 개발 가능성을 검토하였다. Lb. curvatus ML17은 용혈현상을 일으키지 않았으며 ${\beta}$-glucosidase와 ${\beta}$-glucuronidase의 발암효소 활성이 없는 것으로 나타나 균주에 대한 안전성을 확인하였다. 장내 생존성 확인 실험에서는 인공위액에서 2시간 처리한 다음 인공담즙에서 24시간 처리한 결과 초기 균수 대비 12.30%의 생존율을 나타내었다. Caco-2 cell에 대한 부착율은 초기 접종균수가 많을수록 높게 나타났으며, $5.66{\times}10^9$ CFU/mL로 처리 시 17.31%의 부착율을 나타냈는데 20.78%의 부착율을 나타낸 양성 대조군 Lb. rhamnosus GG에 근접하게 높은 부착율을 나타내어 장내환경에서 바람직한 작용을 할 수 있음을 보여주었다. 유해균주에 대한 생육저해 활성 측정에서는 Micrococcus luteus, Bacillus cereus, Salmonella enterica subsp. enterica, Pseudomonas aeruginosa에 대하여 우수한 항균 활성을 나타내었다. DPPH radical 소거능을 이용한 항산화 활성 측정 결과 72.88%로 양성 대조군인 0.1 mg/mL ascorbic acid보다 높은 free radical 소거능을 나타내었다. AGS 위암세포 및 HT-29 결장암세포에 대한 성장 억제 효과에서는 Lb. curvatus ML17의 배양 상등액 $80{\mu}L/mL$ 처리 시 각각 82.91% 및 79.35%의 암세포 성장 억제 효과를 나타내었다. 이와 같은 결과로 Lb. curvatus ML17이 묵은지를 포함한 발효스타터로 사용되었을 경우 식품의 보존성 향상과 품질 유지 및 균주에 의한 건강 기능성을 부여할 수 있으며 식품, 사료, 의약 분야의 다양한 산업적 활용이 기대된다.

• 한국식품과학회지
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• 제42권2호
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• pp.210-216
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• 2010

혈당강하 효과가 있다고 알려진 해당근 추출물을 이용하여 AGEs 생성 저해에 따른 초기 동맥경화증에 대한 효과에 대하여 검토하였다. 먼저 해당근 추출물이 AGEs에 의하여 생성이 촉진된 ROS를 제거하는 효과가 있는지를 확인하기 위하여 항산화 활성을 측정하였다. 총 플라보노이드 함량을 측정한 결과 해당근의 메탄올 추출물은 $128.1{\pm}2.0\;{\mu}g$ quercetin equivalents(QE)/mg DM,총 폴리페놀 함량은 $345.2{\pm}5.7\;{\mu}g$ gallic acid equivalents(GAE)/mg DM으로 나타났다. 환원력 측정을 위한 FRAP assay는 $2.19{\pm}0.1\;mM$ $FeSO_4{\cdot}7H_2O$/mg DM으로 $25.5{\pm}0.3$ $FeSO_4{\cdot}7H_2O$/mg DM 인 ascorbic acid에 비해 약 9% 정도의 환원력을 가지고 있었으며, 전자공여능의 경우 $DPPHSC_{50}$값이 $34.2{\pm}0.1\;{\mu}g$ DM/mL로 대조군인 ascorbic acid의 $6.8{\pm}0.1\;{\mu}g$ DM/mL과 비교하여 약 20% 정도의 높은 항산화력을 갖고 있었다. 또한, HPLC 분석을 통해 해당근 생물 100 g을 기준으로 240 mg의 EGCG와 50 mg의 kaempferol을 함유하는 것으로 나타났다. AGEs에 의해 유도된 HUVEC의 ROS 생성 저해 효과는 해당근 추출물 100, $500\;{\mu}g/mL$ 처리 시 각각 63, 77%로 나타났다. 또한, monocyte adherent assay에서 해당근 추출물은 $100\;{\mu}g/mL$ 처리시 정상을 0%로 하였을 때 AGEs에 의해 유도된 단핵구 부착을 33%, $500\;{\mu}g/mL$ 처리시 약 75% 정도로 농도 의존적으로 부착을 저해하는 것을 관찰할 수 있었으며, 유도되지 않은 HUVEC의 저항은 $113{\Omega}{\cdot}cm^2$ 에서 glycer-AGEs 처리 시 $88{\Omega}{\cdot}cm^2$로 낮아진 것을 해당근 추출물을 $100\;{\mu}g/mL$ 처리 시 약 $102{\Omega}{\cdot}cm^2$, $500\;{\mu}g/mL$ 처리시 약 $106{\Omega}{\cdot}cm^2$로 저항의 감소를 억제하는 것을 관찰할 수 있었다.

• 동의생리병리학회지
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• 제21권1호
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• pp.199-203
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• 2007

Dental caries is a major worldwide oral disease problem. Streptococcus mutans (S. mutans) plays an important role in the information of dental plaque and it is being noticed as major causative bacteria of dental caries. Therefore, the development of more effective, substantial and safe preventive agents against dental caries is strongly required. In the present study, inhibitory effects of the ethanol extract of the husk of pomegranate (Punica granatum L.) on the growth, acid production, adhesion and water-insoluble glucan synthesis of Streptococcus mutans (S. mutans) were examined. The ethanol extract of pomegranate husk (250 - 4000 ${\mu}$g/ml) significantly lowered the growth of S. mutans in a dose dependent manner. The acid production of S. mutans were inhibited by the presence of ethanol extract of pomegranate husk (500 - 4000 ${\mu}$g/ml) significantly. The ethanol extract of pomegranate husk (5000 - 4000 ${\mu}$g/ml) also significantly lowered the adherence of S. mutans. In water-insoluble glucan synthesis assay, 1000 - 4000 ${\mu}$g/ml of the ethanol extract of pomegranate husk significantly inhibited the formation of water-insoluble glucan. These results suggest that pomegranate husk may inhibit the caries-inducing properties of S. mutans. Further studies are necessary to clarify the active constituents of pomegranate husk responsible for such biomolecular activities.

• 동의생리병리학회지
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• 제22권6호
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• pp.1470-1474
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• 2008

This study was performed to examine the anti-metastatic effect of ethanol extract of Samguikoeui-Tang (SGKE), a formula consisting of four oriental herbs, in highly-metastatic HT1080 human fibrosarcoma cells. SGKE significantly inhibited the adhesion of HT1080 cells to matrigel at nontoxic concentrations in a dose-dependent manner, while it did not exert cytotoxicity against HT1080 cells up to the concentration of 100 ${\mu}g$/ml. Also, SGKE depressed the activity of matrix metalloproteinase-2 (MMP-2) and matrix metalloproteinase-9 (MMP-9) by gelatin zymography. However, SGKE did not affect the mRNA expression of MMP-2 and TIMP-2, an inhibitor of MMP-2, by RT-PCR analysis. In addition, the effect of SGKE on HT1080 cell invasion was determined using Boyden chamber assay. SGKE suppressed the invasion of HT1080 cells in a dose-dependent manner. Taken together, these results suggest that SGKE has an anti-metastatic effect via inhibition of MMP-2 and -9 activities.

• 한국재료학회:학술대회논문집
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• 한국재료학회 2009년도 추계학술발표대회
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• pp.42.1-42.1
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• 2009

In recent years, it has been tried to develop the efficacy and bioactivity of Calcium Phosphate cements(CPC) as injectable bone substitute (IBS) by reinforcing them through varying the amount in its compositions and relative concentrations or adding other additives. In this study, the biocompatibility of are inforced Calcium Phosphate-Calcium Sulfate injectable bone substitute (IBS)containing poly ($\varepsilon$-caprolactone)PCL microspheres was evaluated which consisted of solution chitosan and Na-citrate as liquid phase and tetra calcium phosphate (TTCP), dicalciumphosphate anhydrous (DCPA) powder as the solid phase. The in vitrobiocompatibility of the IBS was done using MTT assay and Cellular adhesion and spreading studies. The in vitro experiments with simulated body fluid (SBF) confirmed the formation of apatite on sample surface after 7 and 14 days of incubation in SBF. SEM images for one cell morphologies showed that the cellular attachment was good. MG-63 cells were found to maintain their phenotype on samples and SEM micrograph confirmed that cellular attachment was well. In vitro cytotoxicity tests by an extract dilution method showed that the IBS was cytocompatible for fibroblast L-929.