• Title/Summary/Keyword: Cell delivery

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Preparation and Characterization of Chitosan-coated PLGA Nanoparticle (키토산이 코팅된 PLGA 나노입자의 제조 및 특성)

  • Yu, Su-Gyeong;Nah, Jae-Woon;Jeong, Gyeong-Won
    • Applied Chemistry for Engineering
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    • v.32 no.5
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    • pp.509-515
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    • 2021
  • In this study, poly lactic-co-glycolic acid (PLGA) nanoparticles (PNP) were prepared through double (w/o/w) emlusion and emulsifying solvent-evaporation technique using PLGA, which has biocompatibility and biodegradability. To maximize stability and bioavailability of the particles, chitosan-coated PLGA nanoparticles (CPNP) were prepared by charge interaction between PNP and chitosan. We demonstrated that CPNP can be utilized as a drug carrier of oral administration. The chemical structure of CPNP was analyzed by 1H-NMR and FT-IR, and all characteristic peaks appeared, confirming that it was successfully prepared. In addition, particle size and zeta potential of CPNP were analyzed using dynamic light scattering (DLS) while morphological images were obtained using transmission electron microscope (TEM). Thermal decomposition behavior of CPNP was observed through thermogravimetric analysis (TGA). In addition, the cytotoxicity of CPNP was confirmed by MTT assay at HEK293 and L929 cell lines, and it was proved that there is no toxicity confirmed by the cell viability of above 70% at all concentrations. These results suggest that the CPNP developed in this study may be used as an oral drug delivery carrier.

Transdermal Permeation of Xanthan Gum Bases on the Water-soluble and Lipophilic Antihyperlipoproteinemic Drugs (수용성과 지용성 항고지단백혈증제에 대한 Xanthan Gum 기재에서의 경피투과)

  • 이석우;임윤택;공승대;황성규;이우윤
    • KSBB Journal
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    • v.16 no.3
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    • pp.253-258
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    • 2001
  • Recently, there were many studies not only to enhance drug delivery effect but to reduce side effect. Drug delivery system(DDS) is able to improve efficiency with decreasing side effect of drug dosage. Among these application fields, DDS is often used as the method of drug dosage into the epidermic skin. We investigated characters of transdermal therapeutic system(TTS) and the skin permeability of that with applying DDS. We investigated the permeation of xanthan gum containing drug in rat skin using borizontal membrane cell model. Permeation properties of materials were investigated for water-soluble drug with oxiniacic acid and also for lipophilic drug with clofibrate. The permeation rate of lipophilic drug was found to be faster than that of water-soluble drug in vitro. The rate differences of both water-soluble drug and lipophilic drug according to drug content were negligible. We used glycerin, PEG 600 and oleic acid as enhancers. These results showed that skin permeation rate of each drug across the composite was mainly dependent on the property of base and chemical property of drug etc.. Proper selection of the polymeric materials which resemble and enhance properties of the delivering drug was found to be important in controlling the skin permeation rate. This result suggests a possible use of natural polymer base as a transdermal delivery system of antihyperlipoproteinemic agent.

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Selection of Early Cleaved Embryos and Optimal Recipients to Improve Efficiency of Pig Cloning

  • Koo, Ok-Jae;Lee, Dong-Won;Kang, Jung-Taek;Kwon, Dae-Kee;Park, Hee-Jung;Park, Sol-Ji;Kim, Su-Jin;Jang, Goo;Lee, Byeong-Chun
    • Journal of Embryo Transfer
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    • v.25 no.4
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    • pp.221-227
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    • 2010
  • Early cleavage is a reliable prognostic tool for successful embryo transfer in assisted reproduction because early cleaved embryo show better pregnancy rate after transfer. There for, preparation of good embryo recipient is important factor to optimize efficiency of pig cloning. The present study was performed to evaluate the effect of early cleavage on the in vivo development of cloned embryos and to analyze breed, parity and estrous synchrony to optimize recipient for pig cloning. In vitro matured porcine oocytes derived from local slaughterhouse and fibroblasts derived from miniature pig fetuses were used for somatic cell nuclear transfer (SCNT). Reconstructed embryos were transferred to recipient pigs on the same day of SCNT or after 1~2 days of in vitro culture for selecting early cleaved embryos. Breed, parity and date of standing estrous of recipients were recorded for analysis. After 25~35 days after embryo transfer pregnancy was diagnosed using ultrasonography, and pregnant recipients were monitored till delivery. Between purebred and crossbred, no significant difference was founded in both pregnancy and delivery rates. However, early cleaved embryos showed significantly higher pregnancy (46.2%) and delivery (12.8%) rates compared to non-selectively transferred group (24.8% and 4.5%, respectively). The results also showed that the recipients showing standing estrous on the same day of SCNT and less than 4 parities were most suitable for pig cloning.

Milling Cutter Selection in Machining Center Using AHP (AHP를 활용한 머시닝센터의 밀링커터 선정)

  • Lee, Kyo-Sun;Park, Soo-Yong;Lee, Dong-Hyung
    • Journal of Korean Society of Industrial and Systems Engineering
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    • v.40 no.4
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    • pp.164-170
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    • 2017
  • The CNC machine tool field is showing a growing trend with the recent rapid development of manufacturing industries such as semiconductors, automobiles, medical devices, various inspection and test equipment, mechanical metal processing equipment, aircraft, shipbuilding and electronic equipment. However, small and medium-sized machining companies that use CNC machine tools are experiencing difficulties in increasingly intense competition. Especially, small companies which are receiving orders from 3rd or 4th venders are very difficult in business management. In recent years, company S experienced difficulty to make product quality and delivery time due to the ignorance of the processing method when manufacturing cooling plate jig made of SUS304 material used for cell phone liquid crystal glass processing. In order to solve these problems, we redesigned the process according to the size of our company and tried to manage all processes with quantified data. In the meantime, we have found that there is a need to improve the cutter process, which accounts for most of the machining process. Therefore, we have investigated the correlation between RPM and FEED of three cutters that have been used in the past. As a result, we found that it is the most urgent problem to solve the roughing process during the cutter operation which occupies more than 70% of the total machining. In order to shorten the machining time and improve the quality in machining of SUS304 cooling plate jig, we select the main factors such as price, tool life, maintenance cost, productivity, quality, RPM, and FEED and use AHP to find the most suitable milling cutter. We also tried to solve the problem of delivery, quality and production capacity which was a big problem of S company through experiment operation with selected cutter tool. As a result, the following conclusions were drawn. First, the most efficient of the three cutters currently available in the machining center has proven to be an M-cutter. Second, although one additional facility was required, it was possible to produce the existing facilities without additional investment by supplementing the lack of production capacity due to productivity improvement. Third, the Company's difficulties in delivery and capacity shortfalls have been resolved. Fourth, annual sales increased by KRW 109 million and profits increased by KRW 32 million annually. Fifth, it can confirm the usefulness of AHP method in corporate decision making and it can be utilized in various facility investment and process improvement in the future.

Effect of Cosmetics Contained Isotonic Water Mimicked Body Fluid on Cell Activities and Skin (생체 모사수 화장품이 세포 활성과 피부에 미치는 효과)

  • Park, Sun Young;Lee, Sung Hoon;Kim, Eun Joo;Choi, So Woong;Kim, Ji Young;Cho, Seong A;Cho, Jun Cheol;Lee, Hae Kwang
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.40 no.2
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    • pp.195-201
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    • 2014
  • Body fluid has been studied for diverse fields like Ringer's solutions, artificial joint fluids, cell growth culture media because it plays a crucial role in controlling body temperature and acts as a solvent for diverse metabolite processes in the body and delivery media of mineral, energy source, hormone, signal and drug from and to cell via blood or lymphatic vessel by osmotic pressure or active uptake. Stratum corneum containing extracellular lipids and NMF (natural moisturizing factor) absorbs atmospheric water residing outside of cells and utilize it to hydrate inside of their own. This process is related to skin barrier function. In this study, we conducted the cell viability test with Cell Bio Fluid $Sync^{TM}$, which mimicks body fluids including amino acids, peptides, and monosaccharides to strengthen skin barrier, and the clinical skin improvement test with cosmetics containing Cell Bio Fluid $Sync^{TM}$. In the cell viability test, HaCaT cell was treated with PBS for 3 hours, followed by the treatment of a cell culture medium (DMEM) and isotonic solution (PBS) and Cell Bio Fluid $Sync^{TM}$ for 3 hours each. Then, MTT assay and image analysis were conducted. In the clinical skin improvement test, twenty-one healthy women participated. Participants applied cosmetics containing Cell Bio Fluid $Sync^{TM}$ on their face for a week and evaluated the skin hydration, skin roughness, brightness and evenness. All measurements were conducted after they washed off their face and took a rest under the constant temperature ($22{\pm}2^{\circ}C$) and constant humidity conditions ($50{\pm}5%$) for 20 minutes. All the data were analyzed by SPSS (version 21) software program. Results showed that Cell Bio Fluid $Sync^{TM}$ improved both the cell viability and in vivo skin conditions such as skin hydration, roughness, brightness and evenness.

CONSTRUCTION OF RECOMBINANT HSCC-1 ADENOVIRUS VECTOR FOR ORAL CANCER GENE THERAPY (구강암 유전자 치료를 위한 재조합 HSCC-1 아데노바이러스의 개발)

  • Kim, Chang-Hyen;Kim, Jin-Woo;Kim, Myung-Jin;Pyo, Sung-Woon
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.27 no.2
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    • pp.103-109
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    • 2005
  • In spite of the ongoing advances, standard therapies for oral cancer still has some limitations in efficacy and in ability to prolong survival rate of advanced disease and result in significant functional defect and severe cosmetic deformity. Currently gene therapy using tumor suppressor gene is considered as a potent candidate for new therapeutic approaches that can improve efficacy and reduce complications. The purpose of this research is to identify the role of adenoviral vector to transfer HCCS-1 tumor suppressor gene in oral cancer cells and to find out whether there is a possibility for it to serve in the field of gene therapy. The human SCC-25 cell line was used for transfection. To determine the efficiency of the adenovirus as a gene delivery vector cell line was transduced with LacZ gene and analysed with X-gal staining. Northern blot was performed to confirm the tranfection with HSCC-1 gene and cell viability was assessed by cell cytotoxicity assay. We had successfully construct the recombinant HSCC-1 adenovirus(Ad5CMV-HCCS-1). DNA extracted from Ad5CMV-HCCS-1 revealed HCCS-1 gene is incorporated. The transduction efficiencies were over than 50% of SCC-25 cells with a MOI of 2 and over 95% with a MOI of 50. Northern blot analysis showed that a single 0.6kb mRNA transcript was expressed in Ad5CMV-HCCS-1 transduced SCC-25 cells. There was no or very low transcription HCCS-1 mRNA in wild and Ad5CMV-LacZ transduced SCC-25 cells. Cells transduced with Ad5CMV-HCCS-1 showed significant growth inhibition. By day 6, Ad5CMV-HCCS-1 treated cell count was decreased to 30% of mock-infected cells, while that of Ad5CMV-LacZ treated cells was 90% of mock-infected cells (p<0.05). Finally, these result suggest that the Ad5CMV-HCCS-1 has potential as a gene therapy tool for oral cancer.

Human Organic Anion Transporting Polypeptide 1B3 Applied as an MRI-Based Reporter Gene

  • Song-Ee Baek;Asad Ul-Haq;Dae Hee Kim;Hyoung Wook Choi;Myeong-Jin Kim;Hye Jin Choi;Honsoul Kim
    • Korean Journal of Radiology
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    • v.21 no.6
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    • pp.726-735
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    • 2020
  • Objective: Recent innovations in biology are boosting gene and cell therapy, but monitoring the response to these treatments is difficult. The purpose of this study was to find an MRI-reporter gene that can be used to monitor gene or cell therapy and that can be delivered without a viral vector, as viral vector delivery methods can result in long-term complications. Materials and Methods: CMV promoter-human organic anion transporting polypeptide 1B3 (CMV-hOATP1B3) cDNA or CMV-blank DNA (control) was transfected into HEK293 cells using Lipofectamine. OATP1B3 expression was confirmed by western blotting and confocal microscopy. In vitro cell phantoms were made using transfected HEK293 cells cultured in various concentrations of gadoxetic acid for 24 hours, and images of the phantoms were made with a 9.4T micro-MRI. In vivo xenograft tumors were made by implanting HEK293 cells transfected with CMV-hOATP1B3 (n = 4) or CMV-blank (n = 4) in 8-week-old male nude mice, and MRI was performed before and after intravenous injection of gadoxetic acid (1.2 µL/g). Results: Western blot and confocal microscopy after immunofluorescence staining revealed that only CMV-hOATP1B3-transfected HEK293 cells produced abundant OATP1B3, which localized at the cell membrane. OATP1B3 expression levels remained high through the 25th subculture cycle, but decreased substantially by the 50th subculture cycle. MRI of cell phantoms showed that only the CMV-hOATP1B3-transfected cells produced a significant contrast enhancement effect. In vivo MRI of xenograft tumors revealed that only CMV-hOATP1B3-transfected HEK293 tumors demonstrated a T1 contrast effect, which lasted for at least 5 hours. Conclusion: The human endogenous OATP1B3 gene can be non-virally delivered into cells to induce transient OATP1B3 expression, leading to gadoxetic acid-mediated enhancement on MRI. These results indicate that hOATP1B3 can serve as an MRI-reporter gene while minimizing the risk of long-term complications.

The Effects of OJaJiHwangEumJa(OJJHEJ) Hot water extract & Ultra-fine Powder on Proinflammatory Cytokine of Microglia and Memory Deficit Model (오자지황음자(五子地黃飮子) 열수추출물과 초미세분말이 싸이토카인과 건망증 생쥐모델 기억력감퇴에 미치는 영향)

  • Kim, Seok-Hwan;Lee, Sang-Ryong
    • Journal of Oriental Neuropsychiatry
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    • v.19 no.3
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    • pp.55-68
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    • 2008
  • Background: Microglia produces a barrage of factors (IL-l, TNF-$\alpha$, NO, superoxide) that are toxic to neurons and playa major role in the cellular immune response associated with the pathology of Alzheimer's disease(AD). OJaJiHwangEumJa(OJJHEJ) has been usually used for the treatment of senile disorders. For enhancing efficacy and convenience, the change of the drug delivery device of oriental herbal medicine is required. Objective: This experiment was designed to investigate the effect of the OJJHEJ hot water extract & ultra-fine powder on proinflammatory cytokine of microglia and memory deficit model. Method: The effects of the OJJHEJ hot water extract on production of IL-1$\beta$, IL-6, TNF-$\alpha$, in BV2 microglial cell line treated by lipopolysacchaide(LPS) were investigated. The effects of the OJJHEJ hot water extract & ultra-fine powder on the behavior of the memory deficit mice induced by scopolamine and AChE in serum of the memory deficit mice induced by scopolamine were investigated. Results: 1. The OJJHEJ hot water extract suppressed the production of IL-1$\beta$, IL-6, TNF-$\alpha$ in BV2 microglial cell line and the production of IL-6 was suppressed significantly. 2. The OJJHEJ hot water extract & ultra-fine powder decreased AChE significantly in the serum of the memory deficit mice induced by scopolamine. 3. The OJJHEJ hot water extract & ultra-fine powder groups showed significantly inhibitory effect on the scopolamine-induced impairment of memory in the experiment of Morris water maze. Conclusions: This experiment shows that the OJJHEJ hot water extract & ultra-fine powder might be effective for the prevention and treatment of memory impairment diseases. Investigation into the clinical use of the OJJHEJ hot water extract & ultra-fine powder for Alzheimer's disease is suggested for future research.

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Osteogenic Differentiation of Bone Marrow Stem Cell using Bi-phase Alginate Scaffold Including BMP-2 (BMP-2를 함유한 2상 알지네이트 담체를 이용한 골수줄기세포의 골분화)

  • Lim, Hyun-Ju;Kim, Hak-Tae;Oh, Eun-Jung;Kim, Tae-Jung;Ghim, Han-Do;Choi, Jin-Hyun;Chung, Ho-Yun
    • Archives of Plastic Surgery
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    • v.37 no.3
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    • pp.207-212
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    • 2010
  • Purpose: The object of this study is to develop a novel BMP-2 delivery system for continuous osteogenic differentiation and to induce osteogenesis of stem cells using a bi-phase alginate carrier in vitro. Methods: Alginate nanoparticle loaded BMP-2 was prepared by the reverse emulsification-diffusion technique. Physical properties and release profiles of alginate carriers were measured by Instron and ELISA kit, respectively. Cell viability and alkaline phosphate activity of hBMSCs differentiation was also evaluated by MTS and Metra BAP assays, respectively. Results: Optimal concentration for bi-phase alginate carrier was determined as 2 wt% by evaluating mechanical and biological properties, and differentiation of BMSCs for bone regeneration. The 2% bi-phase alginate carrier had the lowest initial and final release ratio. In addition, the 2% bi-phase alginate carrier had a little higher ALP activity than the homogeneous carrier. An improved controlled release profile was obtained by combining alginate hydrogel with lyophilized particles. Conclusion: Bi-phase alginate carrier has many advantages such as biocompatibility and controlled release capability. It is expected to be effective as a scaffold and carrier in bone tissue engineering.

Liposome/Tat Complex for Facilitating Genistein Uptake into B16 Melanoma Cells

  • Park, Young-Mi;Kang, Myung-Joo;Moon, Ki-Young;Park, Sang-Han;Kang, Mean-Hyung;Choi, Young-Wook
    • Journal of Pharmaceutical Investigation
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    • v.41 no.4
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    • pp.205-210
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    • 2011
  • Genistein (GT), a major isoflavone found in soybeans, has a potent antioxidant effect that protects the skin from UV-induced damages and malignant melanoma. In order to enhance the cellular uptake of GT, liposome/Tat complexes were prepared by an electrostatic interaction of anionic liposome (DMPC/DCP, 9:1 in molar ratio) with Tat peptide (0.02 to 0.08 mole), one of the well-known cell penetrating peptide (CPP). As the amount of Tat increased, the size increased but the zeta potential decreased. In vitro release study with dialysis membrane elicited GT release from liposomal preparations in a controlled manner. The addition of Tat increased GT release, especially for the initial period. In the cellular uptake study by incubating B16 melanoma cells with various liposomal preparations containing GT, B16 melanoma cells demonstrated a time-dependent increase of drug accumulation. Compared to the aqueous GT suspension, intracellular uptake was substantially enhanced by anionic liposomal formulation and further increased by the complex formulation. Therefore, liposome/ Tat complex might be a good candidate for facilitating intracellular drug delivery.