• Title/Summary/Keyword: Cell encapsulation

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Subcellular Responses in Nonhost Plant Infected with Pathogenic and Non-pathogenic Strains of Xanthomonas axonopodis pv. glycines

  • Jeong, Yong-Ho;Kim, Jung-Gun;Chang, Sung-Pae;Hwang, In-Gyu;Kim, Young-Ho
    • The Plant Pathology Journal
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    • v.18 no.3
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    • pp.115-120
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    • 2002
  • Xanthomonas axonopodis pv. glycines, the causal agent of bacterial pustule of soybean, induces hypersensitive response (HR) in a non-host plant, hot pepper (Capsicum annuum). A wild-type strain (8ra) and its non-patho-genic mutant (8-13) of X. axonopodis pv. glycines were inoculated into the pepper leaf tissues and their subcellular responses to the bacterial infections were examined by electron microscopy. Intrastructural changes related to HR were found in the leaf tissues infected with 8ra from 8 h after inoculation, characterized by separation of plasmalemma from the cell wall, formation of small vacuoles and vesicles, formation of cell wall apposition, and cellular necrosis. No such responses were observed in the tissues infected with the mutant. In 8ra, the bacterial cells were attached to the cell walls, with the cell wall material dissolved into and appearing to encapsulate the bacterial cells. The bacterial cells later became entirely embedded in the cell wall material. On the other hand, in 8-13, the bacterial cells were usually not attached tightly to the plant cell wall, and no or poor encapsulation of the bacteria by the wall material occurred, although these were encircled by rather loose wall materials at the later stages.

Enhancement of Photocurrent Generation by C60-encapsulated Single-walled Carbon Nanotubes in Ru-sensitized Photoelectrochemical Cell

  • Lee, Jung-Woo;Park, Tae-Hee;Lee, Jong-Taek;Jang, Mi-Ra;Lee, Seung-Jin;Kim, Hee-Su;Han, Sung-Hwan;Yi, Whi-Kun
    • Bulletin of the Korean Chemical Society
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    • v.33 no.8
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    • pp.2689-2693
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    • 2012
  • Single-walled carbon nanotubes (SWNTs) and $C_{60}$-encapsulated SWNTs ($C_{60}@SWNTs$) are introduced to Ru-sensitized photoelectrochemical cells (PECs), and photocurrents are compared between two cells, i.e., an $RuL_2(NCS)_2$/DAPV/SWNTs/ITO cell and an $RuL_2(NCS)_2$/DAPV/$C_{60}@SWNTs$/ITO cell. [L = 2,2'-bipyridine-4,4'-dicarboxylic acid, DAPV = di-(3-aminopropyl)-viologen, and ITO = indium-tin oxide] The photocurrents are increased by 70.6% in the presence of $C_{60}@SWNTs$. To explain the photocurrent increase, the reverse-field emission method is used, i.e., $RuL_2(NCS)_2$/DAPV/SWNTs/ITO cell (or $RuL_2(NCS)_2$/DAPV/$C_{60}@SWNTs$/ITO cell) as an anode and a counter electrode Pt as a cathode in the external electric field. The improved field emission properties, i.e., ${\beta}$ (field enhancement factor) and emission currents in the reverse-field emission with $C_{60}@SWNTs$ indicate the enhancement of the PEC electric field, which implies the improvement of the electron transfer rate along with the reduced charge recombination in the cell.

Improved Antitumor Efficacy of Hyaluronic Acid-Complexed Paclitaxel Nanoemulsions in Treating Non-Small Cell Lung Cancer

  • Kim, Joo-Eun;Park, Young-Joon
    • Biomolecules & Therapeutics
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    • v.25 no.4
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    • pp.411-416
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    • 2017
  • Paclitaxel (PTX) is a effectively chemotherapeutic agent which is extensively able to treat the non-small cell lung, pancreatic, breast and other cancers. But it is a practically insoluble drug with water solubility less than $1{\mu}g/mL$, which restricts its therapeutic application. To overcome the problem, hyaluronic acid-complexed paclitaxel nanoemulsions (HPNs) were prepared by ionic complexation of paclitaxel (PTX) nanoemulsions and hyaluronic acid (HA) to specifically target non-small cell lung cancer. HPNs were composed of ${\small{DL}}-{\alpha}$-tocopheryl acetate, soybean oil, polysorbate 80, ferric chloride, and HA and fabricated by high-pressure homogenization. The HPNs were $85.2{\pm}7.55nm$ in diameter and had a zeta potential of $-35.7{\pm}0.25mV$. The encapsulation efficiency was almost 100%, and the PTX content was 3.0 mg/mL. We assessed the in vivo antitumor efficacy of the HPNs by measuring changes in tumor volume and body weight in nude mice transplanted with CD44-overexpressing NCI-H460 xenografts and treated with a bolus dose of saline, $Taxol^{(R)}$, PTX nanoemulsions (PNs), or HPNs at a dose of 25 mg/kg. Suppression of cancer cell growth was higher in the PN- and HPN-treated groups than in the $Taxol^{(R)}$ group. In particular, HPN treatment dramatically inhibited tumor growth, likely because of the specific tumor-targeting affinity of HA for CD44-overexpressed cancer cells. The loss of body weight and organ weight did not vary significantly between the groups. It is suggest that HPNs should be used to effective nanocarrier system for targeting delivery of non-small cell lung cancer overexpressing CD44 and high solubilization of poorly soluble drug.

Inhibitory Effects of β-Cyclodextrin-Helenalin Complexes on H-TERT Gene Expression in the T47D Breast Cancer Cell Line - Results of Real Time Quantitative PCR

  • Ghasemali, Samaneh;Nejati-Koshki, Kazem;Akbarzadeh, Abolfazl;Tafsiri, Elham;Zarghami, Nosratollah;Rahmati-Yamchi, Mohamad;Alizadeh, Effat;Barkhordari, Amin;Tozihi, Majid;Kordi, Shirafkan
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.11
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    • pp.6949-6953
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    • 2013
  • Background: Nowadays, the encapsulation of cytotoxic chemotherapeutic agents is attracting interest as a method for drug delivery. We hypothesized that the efficiency of helenalin might be maximized by encapsulation in ${\beta}$-cyclodextrin nanoparticles. Helenalin, with a hydrophobic structure obtained from flowers of Arnica chamissonis and Arnica Montana, has anti-cancer and anti-inflammatory activity but low water solubility and bioavailability. ${\beta}$-Cyclodextrin (${\beta}$-CD) is a cyclic oligosaccharide comprising seven D-glucopyranoside units, linked through 1,4-glycosidic bonds. Materials and Methods: To test our hypothesis, we prepared ${\beta}$-cyclodextrin-helenalin complexes to determine their inhibitory effects on telomerase gene expression by real-time polymerase chain reaction (q-PCR) and cytotoxic effects by colorimetric cell viability (MTT) assay. Results: MTT assay showed that not only ${\beta}$-cyclodextrin has no cytotoxic effect on its own but also it demonstrated that ${\beta}$-cyclodextrin-helenalin complexes inhibited the growth of the T47D breast cancer cell line in a time and dose-dependent manner. Our q-PCR results showed that the expression of telomerase gene was effectively reduced as the concentration of ${\beta}$-cyclodextrin-helenalin complexes increased. Conclusions: ${\beta}$-Cyclodextrin-helenalin complexes exerted cytotoxic effects on T47D cells through down-regulation of telomerase expression and by enhancing Helenalin uptake by cells. Therefore, ${\beta}$-cyclodextrin could be superior carrier for this kind of hydrophobic agent.

Effect of Podophyllotoxin Conjugated Stearic Acid Grafted Chitosan Oligosaccharide Micelle on Human Glioma Cells

  • Wang, Geng Huan;Shen, He Ping;Huang, Xuan;Jiang, Xiao Hong;Jin, Cheng Sheng;Chu, Zheng Min
    • Journal of Korean Neurosurgical Society
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    • v.63 no.6
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    • pp.698-706
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    • 2020
  • Objective : To study the physiochemical characteristics of podophyllotoxin (PPT) conjugated stearic acid grafted chitosan oligosaccharide micelle (PPT-CSO-SA), and evaluate the ability of the potential antineoplastic effects against glioma cells. Methods : PPT-CSO-SA was prepared by a dialysis method. The quality of PPT-CSO-SA including micellar size, zeta potential, drug encapsulation efficiency and drug release profiles was evaluated. Glioma cells were cultured and treated with PPT and PPT-CSO-SA. The ability of glioma cells to uptake PPT-CSO-SA was observed. The proliferation of glioma cells was determined by 3-[4, 5-dimethyl-2-thiazolyl]-2, 5-diphenyl-2H-tetrazolium bromide (MTT) assay. The apoptosis and morphology of U251 cells were observed by 4',6-Diamidino-2-phenylindole dihydrochloride (DAPI) dye staining. Cell cycle analysis was performed by flow cytometry. The migration ability of U251 cells was determined by wound healing test. Results : PPT-CSO-SA had nano-level particle size and sustained release property. The encapsulation efficiency of drug reached a high level. The cellular uptake percentage of PPT in glioma cells was lower than that of PPT-CSO-SA (p<0.05). The inhibitory effect of PPT-CSO-SA on glioma cells proliferation was significantly stronger than that of PPT (p<0.05). The morphologic change of apoptosis cell such as shrinkage, karyorrhexis and karyopyknosis were observed. The percentage of U251 cells in G2/M phase increased significantly in the PPT-CSO-SA group compared with PPT group (p<0.05). Compared with the PPT group, the cell migration ability of the PPT-CSO-SA group was significantly inhibited after 12 and 24 hours (p<0.05). Conclusion : PPT-CSO-SA can effectively enhance the glioma cellular uptake of drugs, inhibit glioma cells proliferation and migration, induce G2/M phase arrest of them, and promote their apoptosis. It may be a promising anti-glioma nano-drug.

Preparation of Microspheres Encapsulating a Recombinant TIMP-1 Adenovirus and their Inhibition of Proliferation of Hepatocellular Carcinoma Cells

  • Xia, Dong;Yao, Hui;Liu, Qing;Xu, Liang
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.12
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    • pp.6363-6368
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    • 2012
  • Objective: The study aim was to prepare poly-DL-lactide-poly (PELA) microspheres encapsulating recombinant tissue inhibitors of metalloproteinase-1 (TIMP-1) in an adenovirus to investigate its inhibition on the proliferation of hepatocellular carcinoma cells HepG2. Methods: Microspheres were prepared by encapsulating the recombinant TIMP-1 adenovirus into biodegradable PELA. The particle size, viral load, encapsulation efficiency and in-vitro release were measured. Microspheres were used to infect HepG2 cells, then infection efficiency was examined under a fluorescent microscope and ultrastructural changes assessed by TEM. Expression of TIMP-1 mRNA in HepG2 cells was examined by semi-quantitative RT-PCR and proliferation by MTT and cell growth curve assays. Results: We successfully prepared microspheres encapsulating recombinant TIMP-1 adenovirus with a diameter of $1.965{\mu}m$, an encapsulation efficiency of 60.0%, a viral load of $10.5{\times}10^8/mg$ and approximate 60% of virus release within 120 h, the total releasing time of which was longer than 240 h. The microspheres were confirmed to be non-toxic with blank microspheres. Infected HepG2 cells could stably maintain in-vitro expression of TIMP-1, with significantly effects on biological behaviour Conclusion: PELA microspheres encapsulating a recombinant TIMP-1 adenovirus can markedly inhibit the proliferation of HepG2 cells, which provides an experimental basis for polymer/chemistry-based gene therapy of hepatocellular carcinomas.

Recombinant Human Epidermal Growth Factor (rhEGF)-loaded Solid Lipid Nanoparticles: Fabrication and Their Skin Accumulation Properties for Topical rhEGF Delivery

  • Hwang, Hee-Jin;Han, Sunhui;Jeon, Sangok;Seo, Joeun;Oh, Dongho;Cho, Seong-Wan;Choi, Young Wook;Lee, Sangkil
    • Bulletin of the Korean Chemical Society
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    • v.35 no.8
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    • pp.2290-2294
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    • 2014
  • For the present study, rhEGF was encapsulated into solid lipid nanoparticles (SLNs). The SLNs were prepared by the $W_1/O/W_2$ double emulsification method combined with the high pressure homogenization method and the physical properties such as particle size, zeta-potential and encapsulation efficiency were measured. The overall particle morphology of SLNs was investigated using a transmission electron microscopy (TEM). The percutaneous skin permeation and accumulation property of rhEGF was evaluated using Franz diffusion cell system along with confocal laser scanning microscopy (CLSM). The mean particle size of rhEGF-loaded SLNs was $104.00{\pm}3.99nm$ and the zeta-potential value was in the range of -$36.99{\pm}0.54mV$, providing a good colloidal stability. The TEM image revealed a spherical shape of SLNs about 100 nm and the encapsulation efficiency was $18.47{\pm}0.22%$. The skin accumulation of rhEGF was enhanced by SLNs. CLSM image analysis provided that the rhEGF rat skin accumulation is facilitated by an entry of SLNs through the pores of skin.

Production of Methanol from Methane by Encapsulated Methylosinus sporium

  • Patel, Sanjay K.S.;Jeong, Jae-Hoon;Mehariya, Sanjeet;Otari, Sachin V.;Madan, Bharat;Haw, Jung Rim;Lee, Jung-Kul;Zhang, Liaoyuan;Kim, In-Won
    • Journal of Microbiology and Biotechnology
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    • v.26 no.12
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    • pp.2098-2105
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    • 2016
  • Massive reserves of methane ($CH_4$) remain unexplored as a feedstock for the production of liquid fuels and chemicals, mainly because of the lack of economically suitable and sustainable strategies for selective oxidation of $CH_4$ to methanol. The present study demonstrates the bioconversion of $CH_4$ to methanol mediated by Type I methanotrophs, such as Methylomicrobium album and Methylomicrobium alcaliphilum. Furthermore, immobilization of a Type II methanotroph, Methylosinus sporium, was carried out using different encapsulation methods, employing sodium-alginate (Na-alginate) and silica gel. The encapsulated cells demonstrated higher stability for methanol production. The optimal pH, temperature, and agitation rate were determined to be pH 7.0, $30^{\circ}C$, and 175 rpm, respectively, using inoculum (1.5 mg of dry cell mass/ml) and 20% of $CH_4$ as a feed. Under these conditions, maximum methanol production (3.43 and 3.73 mM) by the encapsulated cells was recorded. Even after six cycles of reuse, the Na-alginate and silica gel encapsulated cells retained 61.8% and 51.6% of their initial efficiency for methanol production, respectively, in comparison with the efficiency of 11.5% observed in the case of free cells. These results suggest that encapsulation of methanotrophs is a promising approach to improve the stability of methanol production.

Microencapsulation of Probiotic Lactobacillus acidophilus KBL409 by Extrusion Technology to Enhance Survival under Simulated Intestinal and Freeze-Drying Conditions

  • Lee, YunJung;Ji, Yu Ra;Lee, Sumi;Choi, Mi-Jung;Cho, Youngjae
    • Journal of Microbiology and Biotechnology
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    • v.29 no.5
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    • pp.721-730
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    • 2019
  • The probiotic Lactobacillus acidophilus KBL409 was encapsulated with alginate (Al) and alginate-chitosan (Al/Chi) through extrusion method. The sizes and zeta potentials of microspheres were measured to confirm encapsulation. To evaluate the protective effect of microspheres against gastrointestinal fluids, all the samples were exposed to simulated gastric fluids (SGFs, pH 1.5) at $37^{\circ}C$ for 1 or 2 h, followed by incubation with simulated intestinal fluids (SIFs, pH 6.5) for 2 h. The mucoadhesive ability of microspheres was evaluated using the intestinal epithelial cell line HT29-MTX. To extend the shelf-life of probiotics, lyoprotectants such as disaccharide and polysaccharide were mixed with free or encapsulated cells during the freeze-drying process. The size of the microspheres demonstrated a narrow distribution, while the zeta potentials of Al and Al/Chi-microspheres were $-17.9{\pm}2.3$ and $20.4{\pm}2.6mV$, respectively. Among all the samples, Al/Chi-encapsulated cells showed the highest survival rate even after exposure to SGF and SIF. The mucoadhesive abilities of Al and Al/Chi-microspheres were higher than 94%, whereas the free L. acidophilus showed 88.1% mucoadhesion. Ten percent of sucrose showed over 80% survival rate in free or encapsulated cells. Therefore, L. acidophilus encapsulated with Al and Al/Chi-microspheres showed higher survival rates after exposure to the gastrointestinal tract and better mucoadhesive abilities than the free cells. Also, sucrose showed the highest protective effect of L. acidophilus during the freeze-drying process.

Development of a Commercial Process for Micro-Encapsulation of Lactic Acid Bacteria Using Sodium Alginate (알긴산 나트륨을 이용한 유산균 캡슐화의 상업화 공정 개발)

  • Kim, Jiyeon;You, Seong-sik
    • Korean Chemical Engineering Research
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    • v.55 no.3
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    • pp.313-321
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    • 2017
  • We aimed to develop commercialization process of encapsulation which is superior in productivity compared to existing methods by using sodium alginate. Also, in the same process, sodium alginate with chitosan was used to encapsulate lactic acid bacteria with the same process and then the viable cell counts of the two encapsulated lactic acid bacteria were compared. As a test result of the fluidized drying process developed by the present researchers, it was found that the drying time was shortened by 15 to 20 hours compared to the freeze drying method, but the number of viable lactic acid bacteria was about 75% as compared with freeze drying. However, considering the cost and time of drying, it can be confirmed that the commercialization process is possible by the fluidized bed drying method. When the number of viable cells of Ca-alginate capsule and Chitosan-alginate capsule were compared, it was confirmed that there were about $1{\times}10^9$ or more bacteria in the former and about $1{\times}10^3$ in the latter. The lactic acid bacterium capsules prepared by the present technique were stable for 96 hours or more at pH 4.65 and 6.01, but disappeared within 1 hour at pH 7.07 and 8.35. This suggests that the disintegration of lactic acid bacteria can be easily occurred in small and large intestine.