• Title/Summary/Keyword: Cell delivery

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Functional Expression of the Internal Rotenone-Insensitive NADH-Quinone Oxidoreductase (NDI1) Gene of Saccharomyces cerevisiae in Human HeLa Cells

  • Seo, Byoung-Boo
    • Journal of Embryo Transfer
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    • v.25 no.1
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    • pp.35-42
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    • 2010
  • Many studies propose that dysfunction of mitochondrial proton-translocating NADH-ubiquinone oxidoreductase (complex I) is associated with neurodegenerative disorders, such as Parkinson's disease and Huntington's disease. Mammalian mitochondrial proton-translocating NADH-quinone oxidoreductase (complex I) consists of at least 46 different subunits. In contrast, the NDI1 gene of Saccharomyces cerevisiae is a single subunit rotenone-insensitive NADH-quinone oxidoreductase that is located on the matrix side of the inner mitochondrial membrane. With a recombinant adeno-associated virus vector carrying the NDI1 gene (rAAV-NDI1) as the gene delivery method, we were able to attain high transduction efficiencies even in the human epithelial cervical cancer cells that are difficult to transfect by lipofection or calcium phosphate precipitation methods. Using a rAAV-NDI1, we demonstrated that the Ndi1 enzyme is successfully expressed in HeLa cells. The expressed Ndi1 enzyme was recognized to be localized in mitochondria by confocal immunofluorescence microscopic analyses and immunoblotting. Using digitonin-permeabilized cells, it was shown that the NADH oxidase activity of the NDI1-transduced HeLa cells were not affected by rotenone which is inhibitor of complex I, but was inhibited by flavone and antimycin A. The NDI1-transduced cells were able to grow in media containing rotenone. In contrast, control cells that did not receive the NDI1 gene failed to survive. In particular, in the NDI1-transduced cells, the yeast enzyme becomes integrated into the human respiratory chain. It is concluded that the NDI1 gene provides a potentially useful tool for gene therapy of mitochondrial diseases caused by complex I deficiency.

Effect of chicken egg yolk antibody on canine parvoviral enteritis in pups (개 파보바이러스성 장염에 대한 난황항체의 예방 및 치료 효과)

  • Oh, Kyung-Eun;Jeoung, Seok-Young;Kim, Bo-Mi;Jang, Sang-Ho;Lee, Nam-Hyung;Cho, Youngjae;Kim, Doo;Choi, Jung Hoon;Hahn, Tae-Wook
    • Korean Journal of Veterinary Research
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    • v.54 no.2
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    • pp.67-73
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    • 2014
  • Preventive and therapeutic effects of egg yolk antibody, immunoglobulin Y (IgY), against canine parvovirus (CPV) was evaluated in 25 pups orally challenged with CPV-2a. Oral administration of IgY using powder, paste and coated paste delivery systems was compared. Each type of IgY was administered orally for 17 days from 3 days before challenge. The group of pups administered coated IgY showed mild symptoms such as a moderate decrease in total white blood cell count, no depression, vomiting and diarrhea when compared with other groups. The overall clinical score of the group of pups administered coated IgY was significantly lower than that of the challenge control group. However, mortality did not differ among groups because not all pups received symptomatic treatment. These results implied that oral treatment of coated IgY could improve therapeutic effects against CPV challenge if pups received symptomatic treatment.

Enhancing Skin Delivery of 5-Aminolevulinic Acid with Transferosome Using Lyso-Phospholipid and Surfactant

  • Han, In-Sook;Kang, Min-Young;Kim, Moon-Kyu;Kim, Jung-Chul
    • Journal of Pharmaceutical Investigation
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    • v.41 no.1
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    • pp.19-24
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    • 2011
  • In order to enhance the clinical efficacy of 5-aminolevulinic acid-induced photodynamic therapy (ALA-PDT), liposomal formulations using bulk hydrogenated phospholipids from soybean were introduced. Three types of lipids, S75-3, S100-3, and SL80-3 were used for formulating ALA. The pH of all the liposomal ALA is 4.5~5.5 and the size is 50~200 nm. All the liposomal formulations gave better ex vivo ALA skin penetration using nude mice skin in Franz cell than free ALA did. Among them, SL80-3 including 22% of lyso-phosphocholine achieved excellent ALA penetration when compared with those of S75-3 and S100-3 which have only 1~2% of lyso-phospholipids. S100-3 showed a little better results than S75-3 did. Addition of humectants (glycerine, propylene glycol, butylene glycol, betaine) in liposomal ALA formulated with SL80-3 produced little enhancing effect in ALA penetration. On the other hand, addition of surfactants (Tween 20, 60, Brij 72, 76, 78) in same liposomal system produced significant increase in ALA penetration. Among them, transferosomal system of lyso-phospholipid, SL80-3 and the surfactant, Brij76 showed the highest ALA penetration. Furthermore, this system also established the highest in vivo PpIX biosynthesis in hairy mice skin of C57BL/6. These results concluded that the transferosome of SL80-3 and Brij76 produced the best results in both ALA penetration and PpIX biosynthesis, and proved good correlation between them.

Kinetics and Biological Function of Transforming Growth Factor-$\beta$ Isoforms in Bovine and Human Colostrum

  • CHUN, SUNG-KI;NAM, MYOUNG-SOO;GOH, JUHN-SU;KIM, WAN-SUP;HAN, YOUNG-HWAN;KIM, PYEUNG-HYEUN
    • Journal of Microbiology and Biotechnology
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    • v.14 no.6
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    • pp.1267-1274
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    • 2004
  • Colostrum contains various kinds of cytokines including TGF-$\beta$ that has potent regulatory effects on cells of the immune system. We compared the levels of TGF-$\beta$1 and TGF-$\beta$2 in bovine and human colostrum. Based on the isoform-specific ELISA, bovine colostrum collected on day 1 post-delivery retained $53.71{\pm}29.55\;ng/ml$ of TGF-$\beta$1 and $40.41{\pm}21.78\;{\mu}g/ml$ of TGF-$\beta$2 (n=4), while in human, $381.45{\pm}158.24\;ng/ml$ of TGF-$\beta$1 and $41.47{\pm}9.63\;ng/ml$ of TGF-$\beta$2 (n=5). Thus, dominant TGF-$\beta$ isoforms were completely opposite between human and bovine colostrum samples. The concentrations of both isoforms declined as lactation proceeded. Biological activities of the colostrum samples were determined using an MV1LU cell line. Consistent with the result from the immunoassay, TGF-$\beta$1 in human and TGF-$\beta$2 in bovine colostrum were responsible for the anti proliferative activity against MV1LU cells. Furthermore, bovine colostrum increased IgA secretion by LPS-stimulated mesenteric lymph node (MLN) cells, and this effect was abrogated by either anti­TGF-$\beta$2 antibody or combined anti-TGF-$\beta$1/$\beta$2 antibody, but not by anti- TGF-$\beta$1 antibody alone. Similarly, TGF-$\beta$2 in bovine colostrum enhanced the Ig germ line (GL) promoter activity, which is the earliest event toward IgA isotype switching. Taken together, these results suggest that TGF-$\beta$ isoforms, differentially expressed in human and bovine colostrum, may promote IgA isotype production in the neonatal intestine.

Intracellular delivery and anti-tumor activity of polyethyleneglycol liposomes containing cationic lipid (양이온성 지질이 포함된 PEG 리포솜의 세포내 이입 및 항암효력 평가)

  • Jung, Soon-Hwa;Kim, Sung-Kyu;Jung, Suk-Hyun;Seong, Ha-Soo;Cho, Sun-Hang;Shin, Byung-Cheol
    • Journal of Pharmaceutical Investigation
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    • v.38 no.3
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    • pp.163-169
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    • 2008
  • Liposomes are spherical vesicles composed of lipid bilayer membranes. However, the conventional liposomes have been found to be plagued by rapid opsonization and taken up by the reticuloendothelial system (RES), resulting in shortened circulation time and limited intracellular uptake to target cell. In this study, polyethyleneglycol-cationic liposomes (PCL) containing cationic lipid and DSPE-mPEG were prepared by thin film cast-hydration method. The PEG liposomes had approximately $97.0{\pm}1.3\;nm$ of mean particle diameter and $-21.7{\pm}1.2\;mV$ of zeta potential value. PCL had $96.4{\pm}1.8\;nm$ of mean particle diameter and $-8.7{\pm}1.1\;mV$ of zeta potential value with a decrease of about 10 mV compared to the PEG liposomes. Loading of model drug, doxorubicin (DOX), in liposomes were carried out by using remote loading method and the loading efficiency of DOX in liposomes was about $95.0{\pm}1.9%$. Intracellular uptake and cytotoxicity of PCL were higher than that of PEG liposomes to murine B16F10 melanoma cells. In addition, anti-tumor activity of PCL was similar to that of PEG liposomes on growth of A549 human lung carcinoma in BALB/c mice. Consequently, PCL modified with cationic lipid may be applicable as anticancer drug carriers that can increase intracellular uptake and therapeutic efficacy.

Transdermal Delivery of Triamcinolone acetonide Gel by Ultrasound (초음파 조사에 의한 트리암시놀론 아세토니드 겔의 피부투과)

  • Song, Kyung-Suk;Kim, Young-Il;Yang, Jae-Heon
    • Journal of Pharmaceutical Investigation
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    • v.32 no.2
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    • pp.87-93
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    • 2002
  • This study is to enhance drug penetration via skin and investigate anti-inflammation effect following adoption of ultrasound. For this goal gel containing triamcinolone was prepared and the skin penetration rate and the change effects of blood plasma ingredients and serum enzyme were investigated. Using Franz type diffusion cell and the skin of hairless mouse, the permeation enhancing effect of ultrasound was tested. After the injury by direct trauma, the blood test was performed by measuring WBC, lymphocyte, and neutrophyl, and by analyzing CPK and LDH. The ultrasound transducer whose technical specification is geometric area(GA) $1.4\;cm^2$, effective radiation area(ERA) $0.8\;cm^2$, and beam non-uniformity ratio(BNR) 6.0 max was used. The influence of frequency having an effect on skin permeation rate was higher in the case of using 1MHz and continuous treatment. The temperature of receptor phase was not influenced in skin permeation by phonophoresis. Skin permeation increase attended by intensity of ultrasound, the permeation of triamcinolone was accelerated at $2.5\;w/cm^2\;than\;1.0\;w/cm^2$. Following muscle injury phonophoretic group the number of WBC, neutrophil and lympholyte were decreased significantly as compared with both control group and ultrasound group. The result of variation of serum CPK and LDH activity conformed to the phonophoretic effect as same pattern with the variation of WBC, neutrophil and lymphocyte.

Combination Therapy for Gliomas Using Temozolomide and Interferon-Beta Secreting Human Bone Marrow Derived Mesenchymal Stem Cells

  • Park, Jae-Hyun;Ryu, Chung Heon;Kim, Mi Jin;Jeun, Sin-Soo
    • Journal of Korean Neurosurgical Society
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    • v.57 no.5
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    • pp.323-328
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    • 2015
  • Objective : Malignant gliomas are the most common primary tumors of the central nervous system and the prognosis of patients with gliomas is poor. The combination of interferon-bata (IFN-${\beta}$) and temozolomide (TMZ) has shown significant additive antitumor effects in human glioma xenograft models. Considering that the poor survival of patients with human malignant gliomas relates partly to the inability to deliver therapeutic agents to the tumor, the tropism of human bone marrow-derived mesenchymal stem cells (MSC) for malignant gliomas can be exploited to therapeutic advantages. We investigated the combination effects of TMZ and MSCs that secrete IFN-${\beta}$ on gliomas. Methods : We engineered human MSCs to secret mouse IFN-${\beta}$ (MSC-IFN-${\beta}$) via adenoviral transduction and confirmed their secretory capacity using enzyme-linked immunosorbent assays. In vitro and in vivo experiments were performed to determine the effects of the combined TMZ and MSC-IFN-${\beta}$ treatment. Results : In vitro, the combination of MSC-IFN-${\beta}$ and TMZ showed significantly enhanced antitumor effects in GL26 mouse glioma cells. In vivo, the combined MSC-IFN-${\beta}$ and TMZ therapy significantly reduced the tumor size and improved the survival rates compared to each treatment alone. Conclusion : These results suggest that MSCs can be used as an effective delivery vehicle so that the combination of MSC-IFN-${\beta}$ and TMZ could be considered as a new option for the treatment of malignant gliomas.

Preparation and Release Profile of N8f-loaded Polylactide Scaffolds for Tissue Engineered Nerve Regeneration (조직공학적 신경재생을 위한 NGF를 함유한 PLA 담체의 제조 및 방출)

  • 전은경;황혜진;강길선;이일우;이종문
    • Polymer(Korea)
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    • v.25 no.6
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    • pp.893-901
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    • 2001
  • We developed the nerve growth factor (NGF) loaded poly (L - lactide) (PLA) scaffolds by means of emulsion freeze drying method to the possibility for the application of the nerve regeneration of spinal cord disease and the degeneration in Alzheimer's disease. The release amount of NGF from NGF loaded PLA scaffold were analyzed over a 4 week period in vitro at phosphate buffered saline (PBS), pH 7.4, at $37^{\circ}C$. It can be observed the open cell pore structure of porous scaffolds and can be easily controlled the pore structure by the controlling of formulation factors resulting in the controlling of the release rate and the release period. The stability of NGF during the preparation of PLA scaffold was evaluated by comparing the released amounts of total NGF, assayed NGF enzyme - linked immunosorbent assay (ELISA). Released NGF has been found to enhance the neurite sprouting and outgrowth from pheochromocytoma (PC-12) cells. These results suggest that the released NGF from NGF loaded PLA scaffold such as conduit type can be very useful for the nerve regeneration in the neural tissue engineering area.

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Preparation, Characterization and Cytotoxicity of Silibinin-Containing Nanoniosomes in T47D Human Breast Carcinoma Cells

  • Amiri, Boshra;Ebrahimi-Far, Meysam;Saffari, Zahra;Akbarzadeh, Azim;Soleimani, Esmaeil;Chiani, Mohsen
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.8
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    • pp.3835-3838
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    • 2016
  • Background: Breast cancer is one of the most frequent cancer types within female populations. Silibinin is a chemotherapeutic agent ative against cancer. Niosomes are biodegradable, biocompatible, safe and effective carriers for drug delivery. Objective:To prepare nanoniosomal silibinin and evaluate its cytotoxicity inthe T-47D breast cancer cell line. Materials and Methods: Niosomes were prepared by reverse phase evaporation of a mixture of span 20, silibinin, PEG-2000 and cholesterol in chloroform and methanol solvent (1:2 v/v). The solvent phase was evaporated using a rotary evaporator and the remaining gel phase was hydrated in phosphate buffer saline. Mean size, size distribution and zeta potential of niosomes were measured with a Zetasizer instrument and then nanoparticles underwent scanning electron microscopy. The drug releasing pattern was evaluated by dialysis and the cytotoxicity of nanoniosomes in T-47D cells was assessed by MTT assay. Results: Particle size, size variation and zeta potential of the niosomal nanoparticles were measured as $178.4{\pm}5.4nm$, $0.38{\pm}0.09$ and $-15.3{\pm}1.3mV$, respectively. The amount of encapsulated drug and the level of drug loading were determined $98.6{\pm}2.7%$ and $22.3{\pm}1.8%$, respectively; released drug was estimated about $18.6{\pm}2.5%$ after 37 hours. The cytotoxic effects of nanoniosome were significantly increased when compared with the free drug. Conclusions: This study finding suggests that silibinin nanoniosomes could serve as a new drug formulation for breast cancer therapy.

An Enhanced Fast Handover Scheme for Real-Time Traffic in IPv6 Based WiBro Network (IPv6기반 와이브로 시스템에서 실시간 트래픽을 위한 개선된 빠른 핸드오버 방안)

  • Jeong, Seok-Jong;Lee, Sung-Kuen;Park, Jin-Woo
    • The Journal of Korean Institute of Communications and Information Sciences
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    • v.33 no.4A
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    • pp.377-386
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    • 2008
  • In this paper, a new handover management scheme has been proposed to reduce handover latency and to support fast handover without packet losses, so that it may be applicable to the wireless mobile Internet system such as IPv6-based WiBro system. To minimize the handover latency in processing of movement detection, we propose the handover management scheme which simplifies the handover message exchanging procedure between mobile subscriber station (MSS) and network by integrating layer 2 and layer 3 handovers efficiently based on the layer2 information. To reduce the processing delay from new care-of-address (NCoA) configuration during handover, we propose that NCoA is created, distributed and managed by new access control router (NACR). In addition, in order to minimize the packet transmission delay and eliminate the packet losses, the proposed scheme employs a crossover router (CR) which is upper network located over PACR and NACR and employs the packet buffering for MSS. The simulation study shows that the proposed scheme achieves loss-free packet delivery and low latency in the environment of narrow overlapped cell area or high velocity of the MSS, comparing the performance with the conventional schemes.