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      • KCI등재

        Cellular Uptake and Cytotoxicity of β-Lactoglobulin Nanoparticles: The Effects of Particle Size and Surface Charge

        하호경,김진욱,이미령,전우진,이원재 아세아·태평양축산학회 2015 Animal Bioscience Vol.28 No.3

        It is necessary to understand the cellular uptake and cytotoxicity of food-grade delivery systems, such as β-lactoglobulin (β-lg) nanoparticles, for the application of bioactive compounds to functional foods. The objectives of this study were to investigate the relationships between the physicochemical properties of β-lg nanoparticles, such as particle size and zeta-potential value, and their cellular uptakes and cytotoxicity in Caco-2 cells. Physicochemical properties of β-lg nanoparticles were evaluated using particle size analyzer. Flow cytometry and confocal laser scanning microscopy were used to investigate cellular uptake and cytotoxicity of β-lg nanoparticles. The β-lg nanoparticles with various particle sizes (98 to 192 nm) and zeta-potential values (–14.8 to –17.6 mV) were successfully formed. A decrease in heating temperature from 70°C to 60°C resulted in a decrease in the particle size and an increase in the zeta-potential value of β-lg nanoparticles. Non-cytotoxicity was observed in Caco-2 cells treated with β-lg nanoparticles. There was an increase in cellular uptake of β-lg nanoparticles with a decrease in particle size and an increase in zeta-potential value. Cellular uptake β-lg nanoparticles was negatively correlated with particle size and positively correlated with zeta-potential value. Therefore, these results suggest that the particle size and zeta-potential value of β-lg nanoparticles play an important role in the cellular uptake. The β-lg nanoparticles can be used as a delivery system in foods due to its high cellular uptake and non-cytotoxicity.

      • SCIESCOPUSKCI등재

        Cellular Uptake and Cytotoxicity of β-Lactoglobulin Nanoparticles: The Effects of Particle Size and Surface Charge

        Ha, Ho-Kyung,Kim, Jin Wook,Lee, Mee-Ryung,Jun, Woojin,Lee, Won-Jae Asian Australasian Association of Animal Productio 2015 Animal Bioscience Vol.28 No.3

        It is necessary to understand the cellular uptake and cytotoxicity of food-grade delivery systems, such as ${\beta}$-lactoglobulin (${\beta}$-lg) nanoparticles, for the application of bioactive compounds to functional foods. The objectives of this study were to investigate the relationships between the physicochemical properties of ${\beta}$-lg nanoparticles, such as particle size and zeta-potential value, and their cellular uptakes and cytotoxicity in Caco-2 cells. Physicochemical properties of ${\beta}$-lg nanoparticles were evaluated using particle size analyzer. Flow cytometry and confocal laser scanning microscopy were used to investigate cellular uptake and cytotoxicity of ${\beta}$-lg nanoparticles. The ${\beta}$-lg nanoparticles with various particle sizes (98 to 192 nm) and zeta-potential values (-14.8 to -17.6 mV) were successfully formed. A decrease in heating temperature from $70^{\circ}C$ to $60^{\circ}C$ resulted in a decrease in the particle size and an increase in the zeta-potential value of ${\beta}$-lg nanoparticles. Non-cytotoxicity was observed in Caco-2 cells treated with ${\beta}$-lg nanoparticles. There was an increase in cellular uptake of ${\beta}$-lg nanoparticles with a decrease in particle size and an increase in zeta-potential value. Cellular uptake ${\beta}$-lg nanoparticles was negatively correlated with particle size and positively correlated with zeta-potential value. Therefore, these results suggest that the particle size and zeta-potential value of ${\beta}$-lg nanoparticles play an important role in the cellular uptake. The ${\beta}$-lg nanoparticles can be used as a delivery system in foods due to its high cellular uptake and non-cytotoxicity.

      • SCIESCOPUSKCI등재

        Preparation, characterization, and cellular uptake of resveratrol-loaded trimethyl chitosan nanoparticles

        Min, Jeong Bin,Kim, Eun Suh,Lee, Ji-Soo,Lee, Hyeon Gyu Korean Society of Food Science and Technology 2018 Food Science and Biotechnology Vol.27 No.2

        The aim of the study was to encapsulate resveratrol (RV) in trimethyl chitosan (TMC) nanoparticles cross-linked with tripolyphosphate (TPP) and/or alginate to achieve controlled release and improved cellular uptake. TMC (degree of quaternization of 78%) was prepared by reacting purified chitosan with iodomethane. Three types of RV-loaded TMC nanoparticles were prepared: TMC-TPP (TP-NPs), TMC-alginate (TA-NPs), and TMC-alginate-TPP (TAP-NPs). TA-NPs and TAP-NPs showed lower particle size and encapsulation efficiency (EE), better distribution, and more sustained release than TP-NPs due to the high molecular weight and viscous property of alginate. Caco-2 cellular uptake of RV was improved by TMC nanoencapsulation, and TP-NPs showed the highest uptake due to its significantly higher EE. Compared with TAP-NPs, TA-NPs with higher positive surface charge showed higher cellular uptake. Moreover, Caco-2 cell growth-inhibiting activity of RV was significantly increased by TMC nanoencapsulation and TP-NPs showed the significantly highest activity with a good agreement with the permeability results.

      • KCI등재

        Preparation, characterization, and cellular uptake of resveratrolloaded trimethyl chitosan nanoparticles

        민정빈,김은서,이지수,이현규 한국식품과학회 2018 Food Science and Biotechnology Vol.27 No.2

        The aim of the study was to encapsulate resveratrol (RV) in trimethyl chitosan (TMC) nanoparticles cross-linked with tripolyphosphate (TPP) and/or alginate to achieve controlled release and improved cellular uptake. TMC (degree of quaternization of 78%) was prepared by reacting purified chitosan with iodomethane. Three types of RV-loaded TMC nanoparticles were prepared: TMC–TPP (TP-NPs), TMC–alginate (TA-NPs), and TMC–alginate– TPP (TAP-NPs). TA-NPs and TAP-NPs showed lower particle size and encapsulation efficiency (EE), better distribution, and more sustained release than TP-NPs due to the high molecular weight and viscous property of alginate. Caco-2 cellular uptake of RV was improved by TMC nanoencapsulation, and TP-NPs showed the highest uptake due to its significantly higher EE. Compared with TAPNPs, TA-NPs with higher positive surface charge showed higher cellular uptake. Moreover, Caco-2 cell growth-inhibiting activity of RV was significantly increased by TMC nanoencapsulation and TP-NPs showed the significantly highest activity with a good agreement with the permeability results.

      • Preparation of HIFU-triggered tumor-targeted hyaluronic acid micelles for controlled drug release and enhanced cellular uptake

        Zheng, S.,Jin, Z.,Han, J.,Cho, S.,Nguyen, V.D.,Ko, S.Y.,Park, J.O.,Park, S. Elsevier 2016 Colloids and surfaces. B, Biointerfaces Vol.143 No.-

        <P>In this study, a novel type of high intensity focused ultrasound (HIFU)-triggered active tumor-targeting polymeric micelle was prepared and investigated for controlled drug release and enhanced cellular uptake. Amphiphilic hyaluronic acid (HA) conjugates were synthesized to form docetaxel loaded micelles in aqueous conditions with high encapsulation efficiencies of over 80%. The micelle sizes were limited to less than 150 nm, and they varied slightly according to the encapsulated drug amount. Modifying the micellar surface modification with polyethylene glycol diamine successfully inhibited premature drug leakage at a certain level, and it can be expected to prolong the circulation time of the particles in blood. In addition, high-intensity focused ultrasound was introduced to control the release of docetaxel from micelles, to which the release behavior of a drug can be tuned. The in-vitro cell cytotoxicity of docetaxel-loaded micelles was verified against CT-26 and MDA-MB-231 cells. The IC50 values of drug loaded micelles to CT-26 and MDA-MB-231 cells were 1230.2 and 870.9 ng/mL, respectively. However, when exposed to HIFU, the values decreased significantly, to 181.9 and 114.3 ng/mL, suggesting that HIFU can enhance cell cytotoxicity by triggering the release of a drug from the micelles. Furthermore, cellular uptake tests were conducted via the quantitative analysis of intracellular drug concentration within CT-26 (CD44 negative), MDA-MB-231 (CD44 positive), and MDA-MB-231 (CD44 blocked), and then imaged with coumarin-6 loaded micelles. The results verified that intracellular drug delivery can be enhanced efficiently via the CD44 receptor-mediated endocytosis of HA micelles. Moreover, HIFU enhanced the cellular uptake behavior by altering the permeability of the cell membrane. It was also able to aid with the extravasation of micelles into the interior of tumors, which will be explained in further research. Therefore, the present study demonstrates that the micelles prepared in this study can emerge as promising nanocarriers of chemotherapeutic agents for controlled drug release and tumor targeting in cancer treatment. (C) 2016 Elsevier B.V. All rights reserved.</P>

      • pH-sensitive PEGylation of RIPL peptide-conjugated nanostructured lipid carriers: design and in vitro evaluation

        Kim, Chang Hyun,Sa, Cheol-Ki,Goh, Min Su,Lee, Eun Seok,Kang, Tae Hoon,Yoon, Ho Yub,Battogtokh, Gantumur,Ko, Young Tag,Choi, Young Wook Dove Medical Press 2018 International journal of nanomedicine Vol.13 No.-

        <P><B>Background</B></P><P>RIPL peptide (IPLVVPLRRRRRRRRC)-conjugated nanostructured lipid carriers (RIPL-NLCs) can facilitate selective drug delivery to hepsin (Hpn)-expressing cancer cells, but they exhibit low stability in the blood. Generally, biocompatible and nontoxic poly(ethylene glycol) surface modification (PEGylation) can enhance NLC stability, although this may impair drug delivery and NLC clearance. To attain RIPL-NLC steric stabilization without impairing function, pH-sensitive cleavable PEG (cPEG) was grafted onto RIPL-NLCs (cPEG-RIPL-NLCs).</P><P><B>Methods</B></P><P>Various types of NLC formulations including RIPL-NLCs, PEG-RIPL-NLCs, and cPEG-RIPL-NLCs were prepared using the solvent emulsification–evaporation method and characterized for particle size, zeta potential (ZP), and cytotoxicity. The steric stabilization effect was evaluated by plasma protein adsorption and phagocytosis inhibition studies. pH-sensitive cleavage was investigated using the dialysis method under different pH conditions. Employing a fluorescent probe (1,1′-dioctadecyl-3,3,3′,3′-tetramethylindocarbocyanine perchlorate [DiI]), in vitro drug delivery capacity of the cPEG-RIPL-NLCs under different pH conditions was also performed on Hpn-expressing SKOV3 cells and 3D-tumor spheroids.</P><P><B>Results</B></P><P>All prepared NLCs showed homogenous dispersion (<220 nm in size) with a negative ZP (−18 to −22 mV), except for positively charged RIPL-NLCs (~10 mV), revealing no significant cytotoxicity in either SKOV3 or RAW 264.7 cell lines. cPEG-RIPL-NLC protein adsorption was 1.75-fold less than that of RIPL-NLCs, and PEGylation significantly reduced the macrophage uptake. PEG detachment from the cPEG-RIPL-NLCs was pH-sensitive and time dependent. At 2 hours incubation, cPEG-RIPL-NLCs and PEG-RIPL-NLCs exhibited comparable cellular uptake at pH 7.4, whereas cPEG-RIPL-NLC uptake was increased over 2-fold at pH 6.5. 3D-spheroid penetration also demonstrated pH-sensitivity: at pH 7.4, cPEG-RIPL-NLCs could not penetrate deep into the spheroid core region during 2 hours, whereas at pH 6.5, high fluorescence intensity in the core region was observed for both cPEG-RIPL-NLC-and RIPL-NLC-treated groups.</P><P><B>Conclusion</B></P><P>cPEG-RIPL-NLCs are good candidates for Hpn-selective drug targeting in conjunction with pH-responsive PEG cleavage.</P>

      • SCISCIESCOPUS

        Modulation of serum albumin protein corona for exploring cellular behaviors of fattigation-platform nanoparticles

        Nguyen, Van Hong,Meghani, Nilesh M.,Amin, Hardik H.,Tran, Thao T.D.,Tran, Phuong H.L.,Park, Chulhun,Lee, Beom-Jin Elsevier 2018 Colloids and Surfaces B Vol.170 No.-

        <P><B>Abstract</B></P> <P>Albumin is the most abundant protein in blood, and is the most frequently identified protein in the protein corona of nanoparticles (NPs). Thus, albumin plays an important role in modulating NPs’ physicochemical properties and bioavailability. In this study, the effect of bovine serum albumin (BSA) on gelatin-oleic nanoparticles’ (GONs) physicochemical properties and cellular uptake were evaluated. Coumarin-6 was used as indicator to track the cellular uptake of GONs. The binding of BSA onto the GON surface increased the size, slightly reduced the negative net charge of the GON, and improved GON stability. The presence of BSA in cell culture media reduced the cellular uptake of BSA-uncoated GONs on human embryonic kidney cells 293 (HEK 293) and human adenocarcinoma alveolar basal epithelial cells (A549) in the media without FBS addition. Pre-coated BSA corona decreased cellular uptake of GONs in A549 cells in the media, with and without supplemented with 10% fetal bovine serum (FBS) but drastically increased cellular uptake on HEK 293 cells. BSA could be used to modulate protein corona as an endogenous ligand in NP design simply by mixing or incubating BSA with NPs before in vivo administration to inhibit or induce cellular uptake in specific cell types.</P> <P><B>Highlights</B></P> <P> <UL> <LI> BSA adsorption increase GON size, reduced GON zeta potential and stabilized GONs. </LI> <LI> A lower cellular uptake of intact GONs was observed at a higher BSA concentration. </LI> <LI> BSA pre-coated GON modified cellular uptake of GONs in complete media. </LI> </UL> </P> <P><B>Graphical abstract</B></P> <P>[DISPLAY OMISSION]</P>

      • SCISCIESCOPUS

        Assessment of the effect of cooking on speciation and bioaccessibility/cellular uptake of arsenic in rice, using <i>in vitro</i> digestion and Caco-2 and PSI cells as model

        Lee, Seul Gi,Kim, Jinhye,Park, Hyunjoon,Holzapfel, Wilhelm,Lee, Kwang-Won Elsevier 2018 Food and chemical toxicology Vol.111 No.-

        <P><B>Abstract</B></P> <P> <I>In vitro</I> digestion/Caco-2 or pig small intestinal epithelium cell line (PSI) uptake models were used to study the bioaccessibility and cellular uptake of arsenic (As) in cooked white rice and brown rice. The arsenite(AsIII), was the predominant species in cooked rice and in its bioaccessible fractions. The percentage of total As bioaccessibility in white rice (75%) was slightly higher (p=0.061) than that in brown rice(66%). However, there was no difference in the inorganic As (iAs) bioaccessibility between white rice (95%) and brown rice (96%). In Caco-2 cell monolayer, total As retention was 7-31%, transport was 4-25%, and uptake (sum of retention and transport) was 16-38%. In PSI cell model, the retention, transport, and uptake of tAs were 10-28%, 14-31%, and 29-50%, respectively. In both cells, the cellular uptake of tAs in brown rice was 1.4-1.5 folds lower (p<0.05) than that of white rice. These results indicate that the cellular uptake of As can be affected by nutritional compositions. These in vitro screening methods can serve as preliminary screens to predict the relative impact in rice matrix having different As species and processing conditions, although more research efforts should be applied to validating the existing <I>in vitro</I> methods</P> <P><B>Highlights</B></P> <P> <UL> <LI> Cooking effect on speciation of arsenic (As) is assessed. </LI> <LI> Digestion/cell model are used for bioaccessibility and cellular uptake of As. </LI> <LI> Arsenic bioaccessibility in GI fractions are higher than that in the G fractions. </LI> <LI> Total As bioaccessibility is lower in the brown rice than white rice. </LI> <LI> Cellular uptake of brown rice is lower than that of white rice. </LI> </UL> </P> <P><B>Graphical abstract</B></P> <P>[DISPLAY OMISSION]</P>

      • Cellular uptake pathway and drug release characteristics of drug‐encapsulated glycol chitosan nanoparticles in live cells

        Park, Sangjin,Lee, So Jin,Chung, Hyunjin,Her, Songwook,Choi, Yongseok,Kim, Kwangmeyung,Choi, Kuiwon,Kwon, Ick Chan Wiley Subscription Services, Inc., A Wiley Company 2010 Microscopy research and technique Vol.73 No.9

        <P><B>Abstract</B></P><P>Herein, we evaluated the cellular uptake pathways of hydrophobically modified glycol chitosan (HGC) nanoparticles as nano‐sized drug carriers using cellular imaging technology. The endocytic pathway of nanocarriers for intracellular drug delivery is of great interest for the design of high efficacy delivery carriers for therapeutic agents. To evaluate the cellular uptake pathways of HGC nanoparticles, HGC was chemically labeled with near infrared (NIR) fluorescence dye, Cy5.5, to visualize the nanoparticle under confocal laser scanning microscopy. The internalization pathways of HGC nanoparticles were evaluated after treatment of specific endocytosis inhibitors. Importantly, HCG nanoparticles showed different cellular uptake efficiency and intracellular fate in cytoplasm according to the internalization pathways. Furthermore, drug distribution also evaluated according to the endocytic pathways after treatment of drug encapsulated HGC nanoparticles. As a model drug, fluorescent photosensitizer, Ce6, was encapsulated into HGC (Ce6‐HGC) nanoparticles and the distribution of Ce6 in cytoplasm was evaluated using confocal laser scanning microscopy. The intracellular drug distribution showed different manner through specific endocytic pathways. The cellular imaging technology is highly useful for evaluation of endocytosis pathways and intracellular fate of drug delivery carrier which are closely related to drug distribution and therapeutic efficacy. Microsc. Res. Tech. 73:857–865, 2010. © 2010 Wiley‐Liss, Inc.</P>

      • KCI등재

        Size-dependent cellular uptake of sodium alginate passivated tin dioxide nanoparticles in triple-negative breast cancer cells

        Chandrasekaran Karthikeyan,Kokkarachedu Varaprasad,Sungjun Kim,Ashok Kumar Jangid,Wonjeong Lee,Abdulrahman Syedahamed Haja Hameed,Kyobum Kim 한국공업화학회 2023 Journal of Industrial and Engineering Chemistry Vol.123 No.-

        The new development of inorganic (IO) nanoparticle (NPs)-based nanomedicines in anticancer therapy isan active area of research. The cellular uptake of IO NPs plays a crucial role in their efficacy as anticanceragents. In this case, IO NPs cellular uptake depends on physical and chemical parameters, including size,shape, and surface modification of the nanoparticles. From the cellular uptake, one of the essentialparameters for small size plays a critical role in the NPs’ due to their ability to passively diffuse acrossthe cell membrane or enter cells through endocytosis. In this study, the inorganic SnO2 (tin dioxide)and SA (sodium alginate) were made into SnO2 (SASnO2) using a simple one-pot green method. Biomedical studies have shown that SASnO2 NPs exhibit greater antibacterial, antioxidant, and anticancerproperties than SnO2 NPs. The prepared SnO2 and SASnO2 NPs were tested against breast cancer cells inanticancer studies. In cellular uptake studies, the smaller size of SASnO2 NPs (19 nm) resulted in highercellular uptake compared to SnO2 NPs (38 nm). The larger surface area of these SASnO2 NPs allows formore contact with biological membranes and internalization (cell uptake) by cancer cells, resulting inenhanced anticancer therapy when using SASnO2 NPs.

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