• Title/Summary/Keyword: tissue-engineering

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Drug Delivery Study on Chitosan Nanoparticles Using Iron Oxide (II, III) and Valine (Iron Oxide(II, III)와 Valine을 이용한 키토산 나노입자의 약물전달 연구)

  • Jang, So-Hyeon;Kang, Ik-Joong
    • Korean Chemical Engineering Research
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    • v.59 no.4
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    • pp.514-520
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    • 2021
  • A drug delivery system (DDS) based on nanoparticles has been used as a mediator to improve the efficacy of a drug by controlling the amount of drug released and delivering it to a target place. Chitosan, which is non-toxic and biodegradable, has good biocompatibility and excellent adsorption, so it can be used as a drug delivery vehicle. Valine, the essential amino acids, helps muscle growth and tissue recovery, and along with other amino acids. It lowers blood sugar levels and increases growth hormone production. In this study, Valine was adsorbed on magnetic chitosan which is capable of drug absorption, and Fe3O4-Valine CNPs was prepared through cross-linking with TPP (Tripolyphosphate). And then absorption and release trends of valine were investigated with the Fe3O4-Valine CNPs. Fe3O4, which has relatively high stability, is used to make the drug carrier magnetic so that the drug can be delivered to a target place. At optimal conditions, the absorption and release tendency of Fe3O4-Valine CNP was confirmed by analyzing by UV-Vis through the Ninhydrin test which is the color reaction of amino acids and by measuring the size of the particles, it was confirmed that it is suitable as a drug carrier.

Experimental Study of Fire Characteristics by Isocyanate Functional Parameter (이소시아네이트 관능기 매개인자에 의한 화재 특성의 실험적 연구)

  • Lee, Jae-Geol;Han, Kyoung-Ho;Jo, Hyung-Won;Yoon, Do-Young
    • Journal of the Korean Institute of Gas
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    • v.26 no.3
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    • pp.27-37
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    • 2022
  • With the recent increase in the spread of ESS (Electric Storage System), the damage to human life and property is also rapidly increasing due to continuous fires caused by ESS. In the manufacture of urethane sandwich panels used in ESS, it is necessary to improve the flame retardant performance. In this study, in order to realize the flame retardant properties of flexible polyurethane foam, the effect of the tissue density of the product due to the change of the isocyanate functional group parameter that changes the physical properties of the product on the fire performance was studied. The product was manufactured by changing the density of the urethane structure, and combustion performance tests, gas toxicity tests, and smoke density tests were performed. As a result, it was confirmed that the total amount of heat released had excellent performance when the isocyanate functional group was high, and had no correlation with the maximum heat release rate. When the value of the isocyanate functional group was 2.7 or more, the collapse of the shape could be prevented. In the gas hazard test, the performance was increased when the isocyanate functional group was relatively high, so a flame retardant for the Char system, which had a dense structure and easy to form a carbonized film, was added. confirmed to be. Therefore, as a result of this study, it is thought that it will be possible to lay the foundation for the development of a flame retardant to replace the cheap urethane sandwich panel used in the past.

Development and Validation of a Deep Learning System for Segmentation of Abdominal Muscle and Fat on Computed Tomography

  • Hyo Jung Park;Yongbin Shin;Jisuk Park;Hyosang Kim;In Seob Lee;Dong-Woo Seo;Jimi Huh;Tae Young Lee;TaeYong Park;Jeongjin Lee;Kyung Won Kim
    • Korean Journal of Radiology
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    • v.21 no.1
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    • pp.88-100
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    • 2020
  • Objective: We aimed to develop and validate a deep learning system for fully automated segmentation of abdominal muscle and fat areas on computed tomography (CT) images. Materials and Methods: A fully convolutional network-based segmentation system was developed using a training dataset of 883 CT scans from 467 subjects. Axial CT images obtained at the inferior endplate level of the 3rd lumbar vertebra were used for the analysis. Manually drawn segmentation maps of the skeletal muscle, visceral fat, and subcutaneous fat were created to serve as ground truth data. The performance of the fully convolutional network-based segmentation system was evaluated using the Dice similarity coefficient and cross-sectional area error, for both a separate internal validation dataset (426 CT scans from 308 subjects) and an external validation dataset (171 CT scans from 171 subjects from two outside hospitals). Results: The mean Dice similarity coefficients for muscle, subcutaneous fat, and visceral fat were high for both the internal (0.96, 0.97, and 0.97, respectively) and external (0.97, 0.97, and 0.97, respectively) validation datasets, while the mean cross-sectional area errors for muscle, subcutaneous fat, and visceral fat were low for both internal (2.1%, 3.8%, and 1.8%, respectively) and external (2.7%, 4.6%, and 2.3%, respectively) validation datasets. Conclusion: The fully convolutional network-based segmentation system exhibited high performance and accuracy in the automatic segmentation of abdominal muscle and fat on CT images.

Effect of 2-D DBP/PLGA Hybrid Films on Attachment and Proliferation of Intervertebral Disc Cells (2차원적 DBP/PLGA 하이브리드 필름이 디스크 세포의 부착과 증식에 미치는 영향)

  • Ko, Youn-Kyung;Jeong, Jae-Soo;Kim, Soon-Hee;Lim, Ji-Ye;Rhee, John-M.;Kim, Moon-Suk;Lee, Hai-Bang;Khang, Gil-Son
    • Polymer(Korea)
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    • v.32 no.2
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    • pp.109-115
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    • 2008
  • Because demineralized bone particle (DBP) contains various bioactive molecules such as cytokines, it is widely used biomaterials in the field of tissue engineering. In this study, we investigated the effect of 2-dimensional DBP/PLGA hybrid films on adhesion, proliferation and phenotype maintenance of intervertebral disc cells. PLGA films incorporated with different amount (0, 10, 20, 40 and 80 wt%) of DBP were prepared by the solvent evaporation method and characterized by scanning election microscopy (SEM). PLGA film has a flat and smooth surface. According to the increase of content of DBP, the surface of DBP/PLGA film exhibited few agglomerates and increased the roughness of the surface. Annulus fibrosus (AF) and nucleus pulposus (NP) cells were cultured on PLGA and DBP/PLGA film surface, and then examined the cell adhesion and proliferation by the cell count and SEM observation. The result of cell count and SEM observation revealed that 10 and 20% DBP in DBP/PLGA films were superior to adhesion and proliferation of both AF and NP cells. We confirmed that specific gene expression of disc cells on DBP/PLGA film based on the cell count result. Disc cells seeded on 20% DBP/PLGA film expressed the gene of type I and II collagen continuously. Therefore, pertinent content of biomaterials could provide more appropriate condition on adhesion and proliferation of cell. And this results may be used as a basic data for the intervertebral disc regeneration using tissue engineering.

Anti-calcification Effects in Decellularized and Variously Fixed Bovine Pericardium (소심낭 절편의 무세포화와 알코올 전처치를 포함한 여러 고정 처리법 시행 후 석회화 경감 효과 관찰 연구)

  • Hwang, Seong-Wook;Kim, Yong-Jin;Kim, Soo-Hwan;Choi, Seung-Hwa
    • Journal of Chest Surgery
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    • v.43 no.3
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    • pp.235-245
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    • 2010
  • Background: Our goal was to evaluate anti-calcification effects of decellularization and diverse fixing methods including preincubation of the bovine pericardium with ethanol. We also assessed changes in mechanical properties. Material and Method: Harvested bovine pericardium was decellularized with 0.25% sodim dodecysulfate and then treated with 5 methods of fixation: (1) 0.5% glutaraldehyde (GA) for 14 days, (2) 0.5% GA for 5 days, 2% GA for 2 days and 0.25% GA for 7 days, (3) 0.5% GA for 5 days, 2% GA for 2 days, 0.25% GA for 7 days, and then 70% ethanol for 2 days, (4) 0.5% GA for 5 days, a mixture of 2% GA and 70% ethanol for 2 days, and 0.25% GA for 7 days, (5) 0.5% GA for 5 days, a mixture of 2% GA, 65% ethanol, and 5% octanediol for 2 days and then 0.25% GA for 7 days. All treated bovine pericardia were tested for histological variables, lipid content, and mechanical properties including tensile strength and thermal stability. A total 10 kinds of differently treated bovine pericardia were implanted into rat subdermis and harvested 8 weeks later. Harvested pericardia were evaluated for calcium content. Result: No protein denaturation was observed microscopically after decellularization. There was a 32% mean decrease in tensile strength index after decellularization in the bovine pericardium group fixed. Octanediol preincubation attenuated the decrease in tensile strength and maintained thermal stability. TG and cholesterol were not affected by decellularization but were decreased by organic solvent. Calcium content was decreased after decellularization, and organic solvent preincubation decreased calcification in the non-decellularized bovine pericardium group. Conclusion: Decellularization and organic solvent preincubation have anti-calcification effects but decellularization may cause mechanical instability. A method of decellularization and fixation that does not cause damage to matrices will be needed for evaluation of the next step in using tissue-engineering for replacement of cardiac valves.

Biological Activities of Calcium Polyphosphate (Calcium polyphosphate의 생물학적 활성도에 관한 연구)

  • Seol, Yang-Jo;Lee, Jae-Il;Lee, Yong-Moo;Lim, Yoon-Tak;Kim, Seok-Young;Ku, Young;Rhyu, In-Chul;Hahm, Byung-Do;Han, Soo-Boo;Choi, Sang-Mook;Chung, Chong-Pyoung
    • Journal of Periodontal and Implant Science
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    • v.30 no.2
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    • pp.213-231
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    • 2000
  • 이 연구의 목적은 다공성의 CPP 내부에 쥐의 장골의 골수에서 유래된 세포를 접종하고 3차 원적으로 배양하여 CPP가 골 형성을 위한 조직공학의 지지체로 적용가능한가를 연구하는 것과 Calcium PolyPhosphate(CPP)의 돌연변이 유발성을 검사하는 것이다. 무수 ($Ca(H_2PO_4)$)를 condensation하여 무결정의 ($Ca(PO_3)$)를 얻고 이를 용융하고 냉각시킨 후 분쇄하여 Calcium polyphosphate(CPP) powder를 얻었다. 다공성의 CPP는 5% $SiO_2$를 첨가하여 sponge 형태로 $450-550{\mu}m$ 소공의 크기를 가지는 것과(CPP-45ppi) $200-300{\mu}m$의 소공의 크기를 가지는 것(CCP-60ppi) 2가지로 제작하였다. 각각의 CPP matrices는 $5mm{\times}5mm{\times}1mm$의 블록 형태로 만들었다. 체중 100g 내외의 백서에서 장골(femur, tibia)을 채취하여 백서의 장골 골수 세포를 분리하여 배양한 후 24well에 CPP block을 넣고 CPP block 당 $10^5$개의 배양한 세포를 접종하였다. 배양 1, 7, 14, 및 21 일째에 각 well에서 trypsin EDTA를 이용하여 2회 반복하여 cell을 분리하였고, 원심분리한 후 hemacytometer로 측정하였다. 또, 45ppi와 60ppi, 그 리 고 Tissue Culture Polystyrene(control group)에 접종, 배양된 세포들의 염기성 인산분해효소활성도를 배양 7, 14, 및 21 일째에 각각 측정하였다. 각 기간별로 배양된 세포-CPP 혼합체내에서 세포의 부착 및 증식과 형성된 조직의 3차원적 형태를 관찰하기 위하여 주사전자현미경하에서의 관찰하였다. CPP의 돌연변이 유발성 검사 (mutagenicity test)를 위해 hypoxanthine-guanine phosphoribosyl transferase(HPRT) assay를 하였다. NIH3T3 cell line과 CHO-K1 cell line으로 각각 $1000{\mu}g/m{\ell}$, $100{\mu}g/m{\ell}$, $10{\mu}g/m{\ell}$ 그리고 $1{\mu}g/m{\ell}$의 CPP 농도에서 측정하였다. 통계적 분석을 위해서 모든 측정은 각군당 4개체 이상 시험하였고, 각 측정값은 평균값${\pm}$표준편차로 나타내었다. 각 군간의 통계적 유의성 검정을 위해서 Analysis of variance(ANOVA)를 이용하였고 Tukey의 방법으로 사후분석을 실시하였다. 제작된 CPP matrices 소공들이 서로간에 연결이 잘 되어있는 형태였다. 두 가지로 제조된 CPP(45ppi와 60ppi) 모두에서 세포의 부착이 잘 일어났고, 부착된 세포의 분열도 잘 일어났다. 2 가지의 CPP 모두에서 7, 14, 21일째의 세포 수는 1일째에 비해 유의성 있게 증가하였다(P<0.01). 3차원적 구조인 Calcium PolyPhosphate에서 배양한 세포는 24well dish(tissue culture polystyrene)에서 평면적으로 배양한 대조군의 세포에서 보다 염기성 인산분해효소 (Alkaline Phosphatase)를 유의성 있게 높게 나타냈다. 주사전자현미경에서 세포-CPP 혼합체를 관찰한 결과, CPP block에 세포들이 잘부착되어 있었고, 시간이 지남에 따라 세포가 여러 층을 형성하면서 뭉치는 현상을 보였다. 또, HPRT assay 결과 , Calcium PolyPhosphate는 돌연변이 유발성을 보이지 않았다. 이상의 결과로 볼 때 CPP에는 세포부착이 잘 일어나고, 지지체 상에서 세포의 분열도 활발하게 일어나므로 골조직을 위한 조직공학의 우수한 지지체가 될 수 있을 것으로 사료된다.

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Tissue Engineered Cartilage Formation on Various PLGA Scaffolds (PLGA 종류와 담체의 형성 방법에 따른 인간의 조직공학적 연골형성)

  • 김유미;임종옥;정호윤;박태인;백운이
    • Journal of Biomedical Engineering Research
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    • v.23 no.2
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    • pp.147-153
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    • 2002
  • The purpose of this study was to evacuate the effect of different types of Poly(lactic-co-glycolic acid) (PLGA) scaffolds on the formation of human auricular and septal cartilages. All of the scaffolds were formed in a tubular shape for potential application for artificial trachea or esophagus with either 110,000 g/mol PLGA. 220,000 g/mol PLGA. or a combination of both. In order to maintain the tubular shape in vivo, two methods were used. One method was inserting polyethylene tube at the center of scaffolds made of 110,000 g/mol PLGA. The other method involved combination of the two different molecular weight PLGA's. The inner surface of tubular shaped scaffold made with 110,000 g/mol PLGA was coated with 220,000 9/mol PLGA to give more mechanical rigidity. Elastic cartilage was taken from the ear of a patient aged under 20 nears old and hyaline cartilage was taken from the nasal septum. The chondrocytes were then isolated. After second passage, the chondrocytes were seeded on the PLGA scaffolds followed by in vitro culture for one week. The cells-PLGA scaffold complex were implanted subcutaneously on the back of nude mice for 8 weeks. The tissue engineered cartilages were separated from nude mice and examined histologically after staining with the Hematoxylin Eosin. The morphology of the scaffolds were examined by scanning electron microscopy. The pores were well formed and uniformly distributed in the various PLGA scaffolds. After 8 weeks in vivo culture, cartilage was well formed with 110,000 g/mol PLGA. however lumen had collapsed. In contrast. a minimal amount of neocartilage was formed with 220,000 g/mol PLGA, while the architecture of scaffold and lumen were well preserved. Elastic cartilage formed more neocartilage than hyaline. Hyaline and elastic neocartilage were well formed on 110,000 g/mol PLGA with the polyethylene tube, exhibiting mature chondrocytes and preservation of the tubular shape. It was found that 110,000 g/mol PLGA was more appropriate for cartilage formation but higher molecular weight polymer was necessary to maintain the three dimensional shape of the scaffold.

Real-Time RT-PCR for Validation of Reovirus Type 3 Safety During the Manufacture of Mammalian Cell Culture-Derived Biopharmaceuticals (세포배양 유래 생물의약품 생산 공정에서 Reovirus Type 3 안전성 검증을 위한 Real-Time RT-PCR)

  • Lee, Dong-Hyuck;Jeong, Hyo-Sun;Kim, Tae-Eun;Oh, Seon-Hwan;Lee, Jung-Suk;Kim, In-Seop
    • Korean Journal of Microbiology
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    • v.44 no.3
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    • pp.228-236
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    • 2008
  • Validation of viral safety is essential in ensuring the safety of mammalian cell culture-derived biopharmaceuticals, because numerous adventitious viruses have been contaminated during the manufacture of the products. Mammalian cells are highly susceptible to Reovirus type 3 (Reo-3), and there are several reports of Reo-3 contamination during the manufacture of biopharmaceuticals. In order to establish the validation system for the Reo-3 safety, a real-time RT-PCR method was developed for quantitative detection of Reo-3 in cell lines, raw materials, manufacturing processes, and final products as well as Reo-3 clearance validation. Specific primers for amplification of Reo-3 RNA was selected, and Reo-3 RNA was quantified by use of SYBR Green I. The sensitivity of the assay was calculated to be $3.2{\times}10^0\;TCID_{50}/ml$. The real-time RT-PCR method was proven to be reproducible and very specific to Reo-3. The established real-time RT-PCR assay was successfully applied to the validation of Chinese hamster ovary (CHO) cell artificially infected with Reo-3. Reo-3 RNA could be quantified in CHO cell as well as culture supernatant. When the real-time RT-PCR assay was applied to the validation of virus removal during a virus filtration process, the result was similar to that of virus infectivity assay. Therefore, it was concluded that this rapid, specific, sensitive, and robust assay could replace infectivity assay for detection and clearance validation of Reo-3.

Effect of Endothelial Cell Growth Factor and Cyclic AMP Increasers on the Proliferation of Human Omental Microvascular Endothelial Cells (사람의 대망미세혈관내피세포 증식에 대한 내피세포성장인자 및 CYCLIC AMP 증가물질의 효과)

  • Kim, Won-Gon;Kim, Jong-Man;Yu, Se-Yeong
    • Journal of Biomedical Engineering Research
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    • v.16 no.4
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    • pp.463-470
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    • 1995
  • Complete prelining of artificial vascular grafts with autologous endothelial cells may be one of the ideal solutions to obtain a nonthrombogenlc blood-contacting surface. To establish an intact endothelial cell monolayer on a prosthetic surface at the time of implantation,a sufficient number of endothelial cells and adequate propagation condition In cell culture are prerequisites. In this experimental study, endothelial cells from microvessels of adult human oriental adipose tissue were enzymatically harvested, and optimal culture conditions for proliferation of the endothelial cells in cell culture were examined. Human oriental adipose tissue was digested with collagenase and endothelial cells were separated from other stromal elements by mesh filtration method. Cultured cells were identified as endothelial cells by immunofluorescent staining for factor VIII-related antigen. Proliferation in usual 20% fetal bovine serum (FBS) medium or medium containing endothelial cell growth factor (ECGF)(5 ng/ml) and heparin (HEP)(1,000 units/ml) were compared,and the effects of adding compounds that increase intracellular cyclic adenosine monophosphate levels, that is,cholera toxin (CT)(1 $\mu\textrm{g}$/ml) and isobutylmethylxanthine (IBMX)(0.2 ml),were also analyzed. In total,following eight media groups were examined. 1) FBS medium + ECGF + HEP, 2) FBS medium + ECGF + HEP+CT, 3) FBS medium+ECGF+HEP+lBMX, 4) FBS medium+ECGF+HEP+CT+ IBMX, 5) FBSmedium, 6) FBS medium +CT, 7) FBS medium + IBMX, 8) FBS medium + CT + IBMX. It was shown that the medium containing ECGF + HEP with or without cholera toxin was most efficient in Stimulating cell proliferation. IBMX was considered to have antagonistic effect to ECGF. Among experimental groups without ECGF and HEP, the addition of cholera toxin and IBMX was shown to significantly potentiate cell proliferation. This results could provide a practical method for use of cultured human endothelial cells for endothelial cell seeding of cardiovascular prosthetic device, particularly in small-diameter vascular grafts.

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Estimating the Carrying Capacity of a Coastal Bay for Oyster Culture -II. The Carrying Capacity of Geoie-Hansan Bay- (굴 양식수역의 환경용량 산정 -II. 거제 · 한산만의 환경용량-)

  • Park Jong Soo;Kim Hyung Chul;Choi Woo Jeung;Lee Won Chan;Kim Dong Myung;Koo Jun Ho;Park Chung Kil
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.35 no.4
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    • pp.408-416
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    • 2002
  • A 3D hydrodynamic-ecological coupled model was applied to estimate carrying capacity in Geoje-Hansan Bay where is one of the most important oyster culturing grounds in Korea. We considered the carrying capacity as the difference between food supply to the oysters and food demand, considering monthly difference of the actual growth. The food supply to the system was determined from the results of the model simulation (tidal exchange and chlorophyll $\alpha$) over the culturing period from September to May of the following year. The food demand was estimated from the food concentration (chlorophyll $\alpha$) multiple the filtration rate of oysters that is considered monthly different growth rate of oysters and food concentration. The values of carrying capacity for the system varied from 6.1 ton/ha (minimum carrying capacity) in february to 14.91 ton/ha (maximum carrying capacity) in April of marketable size oysters (>4 g wet-tissue weight) depending on temporal variations in the food supply. The oyster production calculated from present facilities was 9 ton/ha in wet-tissue weight in Geoje-Hansan Bay. This value corresponded to $60\%$ of maximum carrying capacity of the system. The optimal carrying capacity without negatively affecting on oyster production was 5.5 ton/ha when calculated from annual statistic data and 6.1 ton/ha when determined by this study. These results suggest that it must be reduced $32\%$~$39\%$ of oyster facilities in the system.