• Title/Summary/Keyword: Cell delivery

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Cellular Protective Effect and Liposome Formulation for Enhanced Transdermal Delivery of Persicaria hydropiper L. Extract (여뀌 추출물의 세포 보호 작용과 피부 흡수 증진을 위한 리포좀 제형 연구)

  • Kim, Jung-Eun;Lee, Hye-Jin;Lim, Myoung-Sun;Park, Min-A;Park, Soo-Nam
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.38 no.1
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    • pp.15-31
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    • 2012
  • In our previous studies, the antioxidant, anti-aging, and antibacterial activities of Persicaria hydropipier L. extract, and the moisturizing effect of cream containing P. hydropipier extract were investigated. In this study, the cellular protective effects of P. hydropipier extract and isoquercitrin, main component from P. hydropipier in $^1O_2$-induced photohemolysis of human erythrocytes and ultraviolet B (UVB)-exposed HaCaT cells were investigated. Liposomes such as ethosome and elastic liposome for enhanced transdermal delivery were prepared. Size, loading efficiency, stability, and cumulative permeated amounts of ethosomes and elastic liposomes were evaluated. P. hydropipier extract and isoquercitrin showed more prominent cellular protective effect than (+)-${\alpha}$-tocopherol, known as lipid antioxidant at $5{\mu}g/mL$. P. hydropipier extract didn't show any characteristics of cytotoxicity at $50{\mu}g/mL$. When HaCaT cells were exposed to a single large dose ($400mJ/cm^2$) of UVB, the extract protected the cells against UVB radiation in a concentration dependent manner ($12.5{\sim}50{\mu}g/mL$). Cell viability of HaCaT cells exposed to UVB $400mJ/cm^2$ was increased by treatment with P. hydropipier extract or isoquercitrin from 36 % (cell viability of positve control groups) to 90 % (cell viability of P. hydropipier extract or isoquercitrin- treated groups). The size of 0.04 % P. hydropiper extract loaded ethosomes was 173.0 nm and the loading efficiency was 55.58 %. 0.04 % P. hydropiper extract loaded ethosomes were stable with as monodisperse particles for 1 week. The ethosome exhibited more skin permeability than general liposome and ethanol solution. The optimal ratio of lipid to surfactant ($Tego^{(R)}$ care 450) of 0.1 % P. hydropiper extract loaded elastic liposomes was observed to be 95 : 5. Vesicle size of 0.1 % P. hydropiper extract loaded elastic liposome was 176.5 nm. The deformability index of the elastic liposome was 16.4. The loading efficiency was 68.8 %. The elastic liposome containing P. hydropiper extract showed more skin permeability than liposome without surfactant ($Tego^{(R)}$ care 450).

Effective Method for Extraction of Cell-Free DNA from Maternal Plasma for Non-Invasive First-Trimester Fetal Gender Determination: A Preliminary Study

  • Lim, Ji-Hyae;Park, So-Yeon;Kim, Shin-Young;Kim, Do-Jin;Kim, Mee-Jin;Yang, Jae-Hyug;Kim, Moon-Young;Kim, Min-Hyoung;Han, Ho-Won;Choi, Kyu-Hong;Ryu, Hyun-Mee
    • Journal of Genetic Medicine
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    • v.7 no.1
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    • pp.53-58
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    • 2010
  • Purpose: To find the most effective method for extraction of cell-free DNA (cf-DNA) from maternal plasma, we compared a blood DNA extraction system (blood kit) and a viral DNA extraction system (viral kit) for non-invasive first-trimester fetal gender determination. Materials and Methods: A prospective cohort study was conducted with maternal plasma collected from 44 women in the first-trimester of pregnancy. The cf-DNA was extracted from maternal plasma using a blood kit and a viral kit. Quantitative fluorescent-polymerase chain reaction (QF-PCR) was used to detect the SRY gene and AMEL gene. The diagnostic accuracy of the QF-PCR results was determined based on comparison with the final delivery records. Results: A total of 44 women were tested, but the final delivery record was only obtained in 36 cases which included 16 male-bearing and 20 female-bearing pregnancies. For the blood kit and viral kit, the diagnostic accuracies for fetal gender determination were 63.9% (23/36) and 97.2% (35/36), respectively. Conclusion: In non-invasive first-trimester fetal gender determination by QF-PCR, using a viral kit for extraction of cf-DNA may result in a higher diagnostic accuracy.

Targeting the epitope spreader Pep19 by naïve human CD45RA+ regulatory T cells dictates a distinct suppressive T cell fate in a novel form of immunotherapy

  • Kim, Hyun-Joo;Cha, Gil Sun;Joo, Ji-Young;Lee, Juyoun;Kim, Sung-Jo;Lee, Jeongae;Park, So Youn;Choi, Jeomil
    • Journal of Periodontal and Implant Science
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    • v.47 no.5
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    • pp.292-311
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    • 2017
  • Purpose: Beyond the limited scope of non-specific polyclonal regulatory T cell (Treg)-based immunotherapy, which depends largely on serendipity, the present study explored a target Treg subset appropriate for the delivery of a novel epitope spreader Pep19 antigen as part of a sophisticated form of immunotherapy with defined antigen specificity that induces immune tolerance. Methods: Human polyclonal $CD4^+CD25^+CD127^{lo-}$ Tregs (127-Tregs) and $na\ddot{i}ve$ $CD4^+CD25^+CD45RA^+$ Tregs (45RA-Tregs) were isolated and were stimulated with target peptide 19 (Pep19)-pulsed dendritic cells in a tolerogenic milieu followed by ex vivo expansion. Low-dose interleukin-2 (IL-2) and rapamycin were added to selectively exclude the outgrowth of contaminating effector T cells (Teffs). The following parameters were investigated in the expanded antigen-specific Tregs: the distinct expression of the immunosuppressive Treg marker Foxp3, epigenetic stability (demethylation in the Treg-specific demethylated region), the suppression of Teffs, expression of the homing receptors CD62L/CCR7, and CD95L-mediated apoptosis. The expanded Tregs were adoptively transferred into an $NOD/scid/IL-2R{\gamma}^{-/-}$ mouse model of collagen-induced arthritis. Results: Epitope-spreader Pep19 targeting by 45RA-Tregs led to an outstanding in vitro suppressive T cell fate characterized by robust ex vivo expansion, the salient expression of Foxp3, high epigenetic stability, enhanced T cell suppression, modest expression of CD62L/CCR7, and higher resistance to CD95L-mediated apoptosis. After adoptive transfer, the distinct fate of these T cells demonstrated a potent in vivo immunotherapeutic capability, as indicated by the complete elimination of footpad swelling, prolonged survival, minimal histopathological changes, and preferential localization of $CD4^+CD25^+$ Tregs at the articular joints in a mechanistic and orchestrated way. Conclusions: We propose human $na\ddot{i}ve$ $CD4^+CD25^+CD45RA^+$ Tregs and the epitope spreader Pep19 as cellular and molecular targets for a novel antigen-specific Treg-based vaccination against collagen-induced arthritis.

Site-Specific Recombination by the Integrase MJ1 on Mammalian Cell (동물 세포 내에서 MJ1 인티그라제에 의한 부위 특이적 재조합)

  • Kim, Hye-Young;Yoon, Bo-Hyun;Chang, Hyo-Ihl
    • Microbiology and Biotechnology Letters
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    • v.39 no.4
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    • pp.337-344
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    • 2011
  • Integrase MJ1 from the bacteriophage ${\Phi}FC1$ carries out recombination between two DNA sequences (the phage attachment site, attP and the bacterial attachment site, attB) in NIH3T3 mouse cells. In this study, the integration vector containing attP, attB and the integrase gene MJ, was constructed. The integration mediated by integrase MJ1 in Escherichia coli led to excision of LacZ. Therefore, the frequency of integration was measured by the counting of the white colony, which is detectable on X-Gal plates. The extrachromosomal integration in NIH3T3 mouse cells was monitored by the expression of the green fluorescent protein (GFP) as a reporter. To demonstrate integration mediated integrase MJ1 in NIH3T3 cells, vectors containing attP and attB were co-transfected into NIH3T3 cells. The integration was confirmed by fluorescent microscopy. The expression of GFP was induced in NIH3T3 cells expressing MJ1 without accessory factors. By contrast, the excision mediated by the MJ1 between attR and attL had no effect on the expression of GFP. These results suggest that integrase MJ1 may enable a variety of genomic modifications for research and therapeutic purposes in higher living cells.

Transduced Human Copper Chaperone for Cu,Zn-SOD (PEP-1-CCS) Protects Against Neuronal Cell Death

  • Choi, Soo Hyun;Kim, Dae Won;Kim, So Young;An, Jae Jin;Lee, Sun Hwa;Choi, Hee Soon;Sohn, Eun Jung;Hwang, Seok-Il;Won, Moo Ho;Kang, Tae-Cheon;Kwon, Hyung Joo;Kang, Jung Hoon;Cho, Sung-Woo;Park, Jinseu;Eum, Won Sik;Choi, Soo Young
    • Molecules and Cells
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    • v.20 no.3
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    • pp.401-408
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    • 2005
  • Reactive oxygen species (ROS) contribute to the development of various human diseases. Cu,Zn-superoxide dismutase (SOD) is one of the major means by which cells counteract the deleterious effects of ROS. SOD activity is dependent upon bound copper ions supplied by its partner metallochaperone protein, copper chaperone for SOD (CCS). In the present study, we investigated the protective effects of PEP-1-CCS against neuronal cell death and ischemic insults. When PEP-1-CCS was added to the culture medium of neuronal cells, it rapidly entered the cells and protected them against paraquat-induced cell death. Moreover, transduced PEP-1-CCS markedly increased endogenous SOD activity in the cells. Immunohistochemical analysis revealed that it prevented neuronal cell death in the hippocampus in response to transient forebrain ischemia. These results suggest that CCS is essential to activate SOD, and that transduction of PEP-1-CCS provides a potential strategy for therapeutic delivery in various human diseases including stroke related to SOD or ROS.

Periodontal healing using a collagen matrix with periodontal ligament progenitor cells in a dehiscence defect model in beagle dogs

  • Yoo, Seung-Yoon;Lee, Jung-Seok;Cha, Jae-Kook;Kim, Seul-Ki;Kim, Chang-Sung
    • Journal of Periodontal and Implant Science
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    • v.49 no.4
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    • pp.215-227
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    • 2019
  • Purpose: To histologically characterize periodontal healing at 8 weeks in surgically created dehiscence defects in beagle dogs that received a collagen matrix with periodontal ligament (PDL) progenitor cells. Methods: The bilateral maxillary premolars and first molars in 6 animals were used. Standardized experimental dehiscence defects were made on the buccal side of 3 premolars, and primary culturing of PDL progenitor cells was performed on the molars. Collagen matrix was used as a scaffold and a delivery system for PDL progenitor cells. The experimental sites were grafted with collagen matrix (COL), PDL progenitor cells with collagen matrix (COL/CELL), or left without any material (CTL). Histologic and histomorphometric analyses were performed after 8 weeks. Results: The defect height from the cementoenamel junction to the most apical point of cementum removal did not significantly differ across the CTL, COL, and COL/CELL groups, at $4.57{\pm}0.28$, $4.56{\pm}0.41$, and $4.64{\pm}0.27mm$ (mean ${\pm}$ standard deviation), respectively; the corresponding values for epithelial adhesion were $1.41{\pm}0.51$, $0.85{\pm}0.29$, and $0.30{\pm}0.41mm$ (P<0.05), the heights of new bone regeneration were $1.32{\pm}0.44$, $1.65{\pm}0.52$, and $1.93{\pm}0.61mm$ (P<0.05), and the cementum regeneration values were $1.15{\pm}0.42$, $1.81{\pm}0.46$, and $2.57{\pm}0.56mm$ (P<0.05). There was significantly more new bone formation in the COL/CELL group than in the CTL group, and new cementum length was also significantly higher in the COL/CELL group. However, there were no significant differences in the width of new cementum among the groups. Conclusions: PDL progenitor cells carried by a synthetic collagen matrix may enhance periodontal regeneration, including cementum and new bone formation.

In vitro Anticancer Activity of Paclitaxel Incorporated in Low-melting Solid Lipid Nanoparticles

  • Lee, Mi-Kyung;Yang, Jae-Heon
    • Journal of Pharmaceutical Investigation
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    • v.39 no.3
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    • pp.201-205
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    • 2009
  • Triglyceride solid lipid with medium chain fatty acid, tricaprin (TC), was used as a core matrix of lipid nanoparticles (LN) to solubilize water-insoluble paclitaxel and enhance the stability of nanoparticles by immobilization of incorporated drug in the solid core during storage at low temperature. In the present study, TC-LN containing paclitaxel was prepared by hot melt homogenization method using TC as a core lipid and phospholipids as stabilizers. The particle size of TC-LN containing paclitaxel was less than 200 nm and its zeta potential was around -40 mV. Calorimetric analysis showed TC core could be solidified by freezing and thawing in the manufacturing process in which the hot dispersion should be prepared at elevated temperature and subsequently cooled to obtain solid lipid nanoparticles. The melting transition of TC core was observed at $27.5^{\circ}C$, which was lower than melting point of TC bulk. The particle size of TC-LN remained unchanged when kept at $4^{\circ}C$. Paclitaxel containing TC-LN showed comparable anticancer activity to the Cremophore ELbased paclitaxel formulation against human ovarian (OVCAR-3) and breast (MCF-7) cancer cell lines. Thus, lipid nanoparticles with medium chain solid lipid may have a potential as alternative delivery system for parenteral administration of paclitaxel.

Lipofectamine-2000 Assisted Magnetofection to Fibroblast Cells Using Polyethyleneimine-Fe3O4@SiO2 Nanoparticles

  • Jang, Eue-Soon;Park, Kyeong-Soon
    • Bulletin of the Korean Chemical Society
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    • v.33 no.8
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    • pp.2567-2573
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    • 2012
  • We successfully synthesized $Fe_3O_4@SiO_2$ nanoparticles with ultrathin silica layer of $1.0{\pm}0.5$ nm that polyethyleneimine (PEI) with low molecular weight of 2.0-4.0 kDa was covalently conjugated with the resulting $Fe_3O_4@SiO_2$ nanoparticles by silane coupling reaction. The PEI-$Fe_3O_4@SiO_2$ nanoparticles were further used as gene delivery vector for a human fibroblast cell (IMR-90) line. Gene transfection efficiency of the PEI-$Fe_3O_4@SiO_2$ complexes did not increase remarkably after magnetofection; however, the addition of Lipofectamine 2000 significantly increased the transfection efficiency of the PEI-$Fe_3O_4@SiO_2$ complexes. We believe that the present approach could be utilized for magnetofection as alternative to $Fe_3O_4$ nanoparticles conjugated with the PEI of high molecular weight thanks to its relatively low cytotoxicity and high transfection efficiency.

Synthesis and Characterization of Upconversion Nanoparticles for Cancer Therapy

  • Choe, Seung-Yu;Kim, Bo-Bae;Kim, Eun-Bi;Lee, Seung-U;Jeon, Seon-A;Park, Tae-Jeong
    • Proceedings of the Korean Vacuum Society Conference
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    • 2016.02a
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    • pp.420.2-420.2
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    • 2016
  • Various fields have been paid attention to upconversion nanoparticles (UCNPs) because of its unique optical properties. Moreover, to use the UC luminescent techniques through cell images for identified apoptosis/necrosis of cancer cells have been performed. They have been studied for a versatile biomedical application such as a biosensing tool, or delivery of active forms of medicines inside living cells. UCNPs have distinctive characteristics such as photoluminescence, special emission, low background fluorescence signal and good colloidal stability, which have many advantages compared with the organic dyes and quantum dots. UCNPs have not only a great potential for imaging (UC luminescence) but also therapies (photo-thermal therapy, PTT and photo-dynamic therapy, PDT) in cancer diagnostics. Therefore, we report the enhancement of upconversion red emission in NaYF4:Yb3+,Er3+ nanoparticles, synthesized via solid-state method with the thermal decomposition of trifluoroacetate as precursors and organic solvent at a high boiling point. The UCNPs have an emission in the field of near infrared wavelength, cubic shape and nano-size in length. In this study, we will further investigate it for cancer therapy with NIR optical detection onto the solid substrate.

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Safety Evaluation of Topical Valproate Application

  • Choi, Sun Young;Seop, Song Yi;Hyun, Moo Yeol;Yoo, Kwang Ho;Kim, Beom Joon;Kim, Myeung Nam;Cho, Jae-We
    • Toxicological Research
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    • v.29 no.2
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    • pp.87-90
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    • 2013
  • The potential role of topical valproate (VPA) in hair regrowth has been recently suggested. However, safety reports of VPA as a topical formulation are lacking. Therefore, in the present study, we investigated whether VPA causes skin irritation in humans. We first performed a cell viability test and showed that VPA did not exhibit toxicity toward HaCaT keratinocytes, fibroblasts, and RBL-3H mast cells. We then performed clinical patch test and skin irritation test through transdermal drug delivery with the help of microneedle rollers. No significant findings were obtained in the clinical patch test. In the skin irritation test, only 1 patient showed erythema at 1 hr, but the irritation reaction faded away within a few hours. Erythema and edema were not observed at 24 hr. We concluded that VPA has minimal potential to elicit skin irritation. Therefore, we consider that VPA can safely be applied to human skin.