• Title/Summary/Keyword: Cell delivery

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Development of Transgenic Carrot Oral Vaccine to Protect against Diarrhea of Piglets (자돈 설사병 방지를 위한 경구백신용 형질전환 당근 개발)

  • 이영선;황철호
    • Journal of Plant Biotechnology
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    • v.29 no.4
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    • pp.287-293
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    • 2002
  • We are trying to develop a transgenic carrot with aims of production and delivery of oral vaccine against microbial enteropathogen using a K88ac pilin gene. A K88ac antigen (pilin) gene was isolated by PCR from the K88ac genomic DNA. The pilin gene was constructed in pGA748 and introduced via Agrobacterium tumefaciens to the explants of carrot hypocotyl and then 494 transgenic lines were established. The amounts of the K88ac antigen produced in each of the cell lines were determined by western and two elite cell lines (M1-17, Y14-1) were selected based on higher levels of expression of the antigens as well as rate of cell growth and efficiency of embryogenesis. In order to test an immunization induced by oral administration of the transgenic carrot, serum of the mice fed with the carrot vaccine were tested in ELISA. It tumed out that the mice fed with 3 g of transgenic carrot showed a similar level of antibody compared to those applied with 10 $\mu\textrm{g}$ of the purified recombinant pilin protein. Besides, various clinical responses were measured after challenging with ETEC K88ac strain to the piglets experiencing an oral immunization with the transgenic carrot. The piglets fed with carrot vaccine showed a lower level of diarrhea in fecal score compared to those fed with non-transgenic carrot. A higher level of increase in weight of the piglets fed with the transgenic carrot vaccine was observed comparing to those fed with non-transgenic carrot as control.

Effects of PLGA/Fibrin Scaffolds on Attachment and Proliferation of Costal Cartilage Cells (PLGA/피브린 지지체가 늑연골 세포의 부착과 성장에 미치는 영향)

  • Song, Jeong Eun;Lee, Yujung;Lee, Yun Me;Cho, Sun Ah;Jang, Ji Eun;Lee, Dongwon;Khang, Gilson
    • Polymer(Korea)
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    • v.37 no.2
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    • pp.141-147
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    • 2013
  • Poly(lactide-co-glycolic acid) (PLGA) has been widely used in the drug delivery and tissue engineering applications because of its good mechanical strength and biodegradation profile. However, cell attachment to the scaffold is low compared with that on fibrin although cells can be attached to the polymer surface. In this study, PLGA scaffolds were soaked in cells-fibrin suspension and polymerized with dropping fibrinogen-thrombin solution. Cellular proliferation activity was observed in PLGA/fibrin-seeded costal cartilage cells (CC) on 1, 3, and 7 days using the MTT assay and SEM. The effects of fibrin on the extracellular matrix (ECM) formation were evaluated using CC cell-seeded PLGA/fibrin scaffolds. The PLGA/fibrin scaffolds elicited more production of glycosaminoglycan (GAG) and collagen than the PLGA scaffold. In this study, fibrin incorporated PLGA scaffolds were prepared to evaluate the effects of fibrin on the cell attachment and proliferation in vitro and in vivo. In this result, we confirmed that proliferation of cells in PLGA/fibrin scaffolds were better than in PLGA scaffolds. The PLGA/fibrin scaffolds provide suitable environment for growth and proliferation of costal cartilage cells.

Tissue Engineered Catilage Reconstruction with Alginate Sponge Containing Demineralized Bone Particles (탈미네랄골분이 첨가된 알지네이트 스펀지에서 조직공학적 연골 재건)

  • Kim, Hye Min;Park, Jin Young;Kim, Eun Young;Song, Jeong Eun;Kwon, Soon Yong;Chung, Jin Wha;Khang, Gilson
    • Polymer(Korea)
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    • v.38 no.3
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    • pp.278-285
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    • 2014
  • Demineralized bone particles (DBP) and alginate hybrid sponges were fabricated at 10, 20, 40 and 80% DBP/alginate hybrid ratios for seeding chondrocyte. Cell proliferation was measured via MTT assay. Morphological observation, histology, biological assay and RT-PCR were performed at each time point 1, 2 and 3 weeks. The cell viability was better in 20% DBP/alginate sponges than in other sponges. SEM results showed that more attached and more proliferated cells in the 20% DBP/alginate sponges with the lapse of time. Finally, histochemical assay results showed that the phenotype of chondrocyte was well maintained and both acidic mucopolysaccharide and type II collagen was well formed at 20% sponges. This study suggested that DBP/alginate sponge may serve as a potential cell delivery vehicle and a structural basis for tissue engineered articular cartilage.

Effect of PLGA Scaffold Containing Demineralized Bone Solution for Articular Cartilage Tissue Engineering: In Vitro Test (조직공학적 연골재생을 위한 In Vitro 환경에서의 탈미네랄화 골분용액을 함유한 PLGA 지지체의 효과)

  • Ahn, Woo-Young;Kim, Hye-Lin;Song, Jeong-Eun;Lee, Dong-Won;Khang, Gil-Son
    • Polymer(Korea)
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    • v.35 no.6
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    • pp.499-504
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    • 2011
  • Articular cartilage has an intrinsic difficulty in recovering damages, which requires its tissue engineering treatment. Demineralized bone particle (DBP) contains various bioactive molecules. It is widely used biomaterials in the field of tissue engineering. We developed the synthetic/natural hybrid scaffolds with poly(lactide-co-glycolide) (PLGA) and solution of DBP. The chondrocytes were seeded on the PLGA-DBP scaffolds and MTT assay, morphological observation, biological assay for collagen, sGAG, and RT-PCR were performed to analyze the effect of the DBP on cell viability and extracellular matrix secretion. In SEM observation, we observed that PLGA-DBP scaffolds had uniform porosity. As MTT assay showed scaffolds containing DB solution had higher cell viability then only PLGA scaffolds. The PLGA-DBP scaffolds had better ECM production than PLGA scaffold. It was proven by the higher specific mRNA expression in the PLGA-DBP scaffold than that in PLGA scaffold. These results indicated that PLGA-DBP scaffolds might serve as potential cell delivery vehicles and structural bases for in vitro tissue engineered articular cartilage.

Effectiveness Evaluation of Displacement Accommodatable Pressure Measuring Jig for Quality Assessment of Pressure Application Device (압력 인가 장치의 품질관리를 위한 변위 수용이 가능한 압력 측정용 지그의 유효성 평가)

  • Mun, Chang-Su;Jun, Sung-Chul;Noh, Si-Cheol
    • Journal of the Institute of Convergence Signal Processing
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    • v.21 no.2
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    • pp.61-66
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    • 2020
  • Recently, a variety of electric anesthetics devices have been developed and used in clinical practice to reduce the fatigue of the operator during local anesthesia for dental procedures and to compensate for the disadvantages of manual anesthesia device. In this electric anesthesia injection device, the accurate and constant delivery of pressure for drug infusion is a very important performance factor. In order to evaluate the accuracy of the transfer pressure, a small pressure gauge using a load cell is often used, but since the elastic body inside the load cell may not be able to accommodate a sufficient displacement, an error may occur when evaluating pressure performance. For these reasons, in this study, we proposed and evaluated a silicon-chrome steel (Si-Cr steel) spring jig that can accommodate relatively large displacements that can be used when evaluating the performance of a pressure-controlled pressure application device using a load cell type pressure gauge. As a result of the pressure transmissibility test and repeated measurement results using a commercial dental anesthesia injection device, a more stable result was obtained when using a spring jig, and it was confirmed that the frequency of abnormally high measurement was reduced.

Clinical Study on the Successful Pregnancy and Delivery after Transfer of Human Blastocysts Cryopreserved by Vitrification (인간 포배란의 유리화동결 융해 후 임신 및 분만에 관한 연구)

  • Choi, Dong-Hee;Chung, Hyung-Min;Chung, Mi-Kyung;Lee, Sook-Hwan;Nam, Yoon-Seung;Park, Chan;Kwak, In-Pyung;Yoon, Tae-Ki
    • Clinical and Experimental Reproductive Medicine
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    • v.27 no.4
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    • pp.367-372
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    • 2000
  • Objective: This study was performed to evaluate whether vitrification method could be used for the cryopreservation of human blastocysts derived from IVF program. Methods: Surplus embryos were obtained from consented IVF patients. Controlled ovarian hyperstimulation was done with midluteal GnRH agonist, gonadotropin and hCG. After oocyte retrieval and insemination, fresh embryo transfer was done at $4{\sim}8$ cell stage. The surplus embryos after ET were cultured in blastocyst medium up to 6 days after oocyte retrieval. Obtained blastocysts were cryopreserved with our vitrification method. Blastocysts were exposed to 1.5 Methylene glycol (EG) in phosphate buffered saline (PBS) for 2.5 minutes, followed by 5.5 M EG plus 1 M sucrose for 20 seconds. Then 1 to 3 blastocysts were mounted on electron microscope (EM) grid and the grid was plunged into liquid nitrogen for storage. For thawing, blastocyst-containing EM grids were sequentially transferred in 1.0 M, 0.5 M, 0.25 M, 0.125 M and 0 M sucrose solution at the intervals of2.5 minutes. And blastocysts were cultured for about 6 hours and only re-expanded blastocysts were transferred to uterus of the patients on 4 to 5 days after ovulation in natural cycle or on 18 to 19 day of artificial cycle. Results: From Oct. 1998 to Jul. 1999, 34 patients were agreed to participate in this study. The mean age and duration of infertility of the patients were 31.6 years and 4.1 years, respectively. Among 34 cycles. replacements could be done in 20 cycles (58.8%). A total 93 blastocysts were thawed and 48 (51.6%) of them survived. Thirty-eight blastocysts, mean 1.9 embryos per patient, were transferred, resulting in 5 clinical pregnancies which consisted of 1 triplet, 2 sets of twins and 2 singleton pregnancies. The pregnancy rate per transfer was 25% and implantation rate was 23.6%. Five patients delivered 7 healthy babies including 2 sets of twins at term. Conclusion: Successful pregnancies and deliveries were established after transfer of vitrified human blastocysts. Vitrification using ethylene glycol as cryoprotectant and electron microscope grid is a rapid and simple method that can be effectively applied for the cryopreservation of human blastocysts.

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Zanamivir Oral Delivery: Enhanced Plasma and Lung Bioavailability in Rats

  • Shanmugam, Srinivasan;Im, Ho Taek;Sohn, Young Taek;Kim, Kyung Soo;Kim, Yong-Il;Yong, Chul Soon;Kim, Jong Oh;Choi, Han-Gon;Woo, Jong Soo
    • Biomolecules & Therapeutics
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    • v.21 no.2
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    • pp.161-169
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    • 2013
  • The objective of this study was to enhance the oral bioavailability (BA) of zanamivir (ZMR) by increasing its intestinal permeability using permeation enhancers (PE). Four different classes of PEs (Labrasol$^{(R)}$, sodium cholate, sodium caprate, hydroxypropyl ${\beta}$-cyclodextrin) were investigated for their ability to enhance the permeation of ZMR across Caco-2 cell monolayers. The flux and $P_{app}$ of ZMR in the presence of sodium caprate (SC) was significantly higher than other PEs in comparison to control, and was selected for further investigation. All concentrations of SC (10-200 mM) demonstrated enhanced flux of ZMR in comparison to control. The highest flux (13 folds higher than control) was achieved for the formulation with highest SC concentration (200 mM). The relative BA of ZMR formulation containing SC (PO-SC) in plasma at a dose of 10 mg/kg following oral administration in rats was 317.65% in comparison to control formulation (PO-C). Besides, the $AUC_{0-24\;h}$ of ZMR in the lungs following oral administration of PO-SC was $125.22{\pm}27.25$ ng hr $ml^{-1}$ with a $C_{max}$ of $156.00{\pm}24.00$ ng/ml reached at $0.50{\pm}0.00$ h. But, there was no ZMR detected in the lungs following administration of control formulation (PO-C). The findings of this study indicated that the oral formulation PO-SC containing ZMR and SC was able to enhance the BA of ZMR in plasma to an appropriate amount that would make ZMR available in lungs at a concentration higher (>10 ng/ml) than the $IC_{50}$ concentration of influenza virus (0.64-7.9 ng/ml) to exert its therapeutic effect.

Oral Delivery of Probiotics in Poultry Using pH-Sensitive Tablets

  • Jiang, Tao;Li, Hui-Shan;Han, Geon Goo;Singh, Bijay;Kang, Sang-Kee;Bok, Jin-Duck;Kim, Dae-Duk;Hong, Zhong-Shan;Choi, Yun-Jaie;Cho, Chong-Su
    • Journal of Microbiology and Biotechnology
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    • v.27 no.4
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    • pp.739-746
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    • 2017
  • As alternatives to antibiotics in livestocks, probiotics have been used, although most of them in the form of liquid or semisolid formulations, which show low cell viability after oral administration. Therefore, suitable dry dosage forms should be developed for livestocks to protect probiotics against the low pH in the stomach such that the products have higher probiotics survivability. Here, in order to develop a dry dosage forms of probiotics for poultry, we used hydroxypropyl methylcellulose phthalate 55 (HPMCP 55) as a tablet-forming matrix to develop probiotics in a tablet form for poultry. Here, we made three different kinds of probiotics-loaded tablet under different compression forces and investigated their characteristics based on their survivability, morphology, disintegration time, and kinetics in simulated gastrointestinal fluid. The results indicated that the probiotics formulated in the tablets displayed higher survival rates in acidic gastric conditions than probiotics in solution. Rapid release of the probiotics from the tablets occurred in simulated intestinal fluid because of fast swelling of the tablets in neutral pH. As a matrix of tablet, HPMCP 55 provided good viability of probiotics after 6 months under refrigeration. Moreover, after oral administration of probiotics-loaded tablets to chicken, more viable probiotics were observed, than with solution type, through several digestive areas of chicken by the tablets.

Assessment of Discoidal Polymeric Nanoconstructs as a Drug Carrier (약물 운반체로서의 폴리머 디스크 나노 입자에 대한 평가)

  • BAE, J.Y.;OH, E.S.;AHN, H.J.;KEY, Jaehong
    • Journal of Biomedical Engineering Research
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    • v.38 no.1
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    • pp.43-48
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    • 2017
  • Chemotherapy, radiation therapy, and surgery are major methods to treat cancer. However, current cancer treatments report severe side effects and high recurrences. Recent studies about engineering nanoparticles as a drug carrier suggest possibilities in terms of specific targeting and spatiotemporal release of drugs. While many nanoparticles demonstrate lower toxicity and better targeting results than free drugs, they still need to improve their performance dramatically in terms of targeting accuracy, immune responses, and non-specific accumulation at organs. One possible way to overcome the challenges is to make precisely controlled nanoparticles with respect to size, shape, surface properties, and mechanical stiffness. Here, we demonstrate $500{\times}200nm$ discoidal polymeric nanoconstructs (DPNs) as a drug delivery carrier. DPNs were prepared by using a top-down fabrication method that we previously reported to control shape as well as size. Moreover, DPNs have multiple payloads, poly lactic-co-glycolic acid (PLGA), polyethylene glycol (PEG), lipid-Rhodamine B dye (RhB) and Salinomycin. In this study, we demonstrated a potential of DPNs as a drug carrier to treat cancer.

Cellular Protective Effect and Liposome Formulation for Enhanced Transdermal Delivery of Isoquercitrin (Isoquercitrin의 세포 보호 작용과 피부 흡수 증진을 위한 리포좀 제형 연구)

  • Jo, Na-Rae;Gu, Hyun-A;Park, Su-Ah;Han, Seat-Byeol;Park, Soo-Nam
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.38 no.2
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    • pp.103-118
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    • 2012
  • In this study, the cellular protective effect of isoquercitrin against $H_2O_2$ and rose bengal-indued HaCaT cell damage was investigated. The ethosome and elastic liposome for enhanced transdermal delivery were prepared. Particle size, loading efficiency and cumulative permeated amounts of them were evaluated. Isoquercitrin didn't show any characteristic cytotoxicity at 50 ${\mu}M$. When HaCaT cells were treated with 5 mM $H_2O_2$ and 25 ${\mu}M$ rose bengal, isoquercitrin protected the cells against the oxidative damage in a concentration dependent manner (6.25 ~ 50 ${\mu}M$). The size of 0.03 % isoquercitrin loaded ethosome was 222.85 nm and the loading efficiency was 82.26 %. The ethosome loaded with 0.03 % isoquercitrin was stable and maintained the constant particle size for 2 weeks after being prepared. The ethosome exhibited more enhanced skin permeability than general liposome and ethanol solution. The optimal ratio of lipid to surfactant of 0.1 % isoquercitrin loaded elastic liposomes was observed to be 89 : 5 through evaluating particle size (341.2 nm), deformability index (59.89), loading efficiency (54.3 %), and skin permeability (54.4 %).