• Title/Summary/Keyword: lyophilization

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Fabrication and Characterization of 3-D Porous Collagen Scaffold (3차원 다공성 콜라겐지지체의 제조 및 특성 분석)

  • Kim, Jin-Tae;Lim, Sumin;Kim, Byoung Soo;Lee, Deuk Yong;Choi, Jae Ha
    • Journal of Biomedical Engineering Research
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    • v.35 no.6
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    • pp.192-196
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    • 2014
  • Collagen scaffolds were synthesized by cross linking into a solution mixture of 1-ethyl-3-[3-dimethylaminopropyl] carbodiimide hydrochlorid(EDC) in ethanol, followed by pressing, cleaning and lyophilization process after the type I atelo-collagen solutions in D.I water(pH3). The experimental conditions are collagen concentration of 1.0 wt%, 3.0 wt%, 5.0 wt% and differential concentration of cross-linker. Then, parametric studies were performed by varying the parameters to investigate the morphology, the porosity, the swelling ratio and the thickness and genotoxicity of the scaffolds. The scaffolds thickness pattern was regular to concentration of the degree of cross-linker and collagen. It was observed that the swelling ratio, the degree of crosslink, and the pore size(thickness of scaffold) can be controlled by adjusting the collagen, crosslinker. Among the parameters investigated, the smallest thickness can be achieved by collagen, crosslinker concentrate condition. The collagen scaffold is induced no genotoxicity. The lowest swelling ratio, as an indication of the highest degree of crosslink, can be obtained by adding crosslink agent.

Preparation and Evaluation of Freeze-dried Solid Lipid Nanoparticles with Various Cryoprotectants

  • Li, Ri Hua;Seo, Seung-Yong;Eun, Jae-Soon;Lee, Mi-Kyung
    • Journal of Pharmaceutical Investigation
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    • v.40 no.1
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    • pp.39-43
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    • 2010
  • Solid lipid nanoparticles (SLNs) were freeze-dried to obtain a stable solid dosage form with the aid of various cryoprotectants such as trehalose, sucrose, glucose, fructose, and glycerol. Tricaprin(TC) and trilaurin(TL) were used as lipid matrices for SLNs and stabilizers were egg phosphatidylcholine and pegylated phospholipid. All cryoprotectants tested did not cause changes in mean particle size of SLNs when mixed with SLNs before freeze-drying. However, the mean particle sizes of reconstituted SLNs after freeze-drying were significantly different from those of the un-lyophilized original SLN dispersions depending on the types and concentration of cryoprotectants. Although the freeze-dried SLNs without any cryoprotectants were easily reconstituted by hand-shaking, the mean particle size drastically increased (> $8\;{\mu}m$ for TC SLNs and around $1\;{\mu}m$ for TL SLNs) compared to that of un-lyophilized original dispersion (97 nm for TC SLNs and 164 nm for TL SLNs). Trehalose and sucrose were the most effective additives to protect the SLNs during lyophilization. The reconstituted SLNs were physically stable for 24 hours when lyophilized with 12.5% trehalose, sucrose, glucose, fructose or glycerol.

Three-Dimensional Porous Collagen/Chitosan Complex Sponge for Tissue Engineering

  • Kim, Sung Eun;Cho, Yong Woo;Kang, Eun Jung;Kwon, Ick Chan;Lee, Eunhee Bae;Kim, Jung Hyun;Chung, Hesson;Jeong, Seo Young
    • Fibers and Polymers
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    • v.2 no.2
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    • pp.64-70
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    • 2001
  • A three-dimensional, porous collagen/chitosan complex sponge was prepared to closely simulate basic extracellular matrix (ECM) constitutes, collagen and glycosaminoglycan. The complex sponge was prepared by a lyophilization method and had the regular network with highly porous structure, suitable for cell adhesion and growth. The pores were well interconnected, and their distribution was fairly homogeneous. The complex sponge was crosslinked using 1-ethyl-3-(3-dimethyl aminopropyl) carbodiimide (EDC) and N-hydroxysuccinimide (NHS) to increase its boilogical stability and enhance its mechanical properties. The crosslinking medium has a great effect on the inner structure of the sponge. The homogeneous, porous structure of the sponge was remarkably collapsed in an aqueous crosslinking medium. However, the morphology of the sponge remained almost intact in a water/ethanol mixture crosslinking milieu. Mechanical properties of the collagen/chitosan sponge were significantly enhanced by EDC-mediated crosslinking. The potential of the sponge as a scaffold for tissue engineering was investigated using a Chinese hamster ovary cell (CHO-K1) line.

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Application of TMAH-based Pyrolysis Mass Spectrometry to a Biological Detection System (TMAH에 기반한 열분해 질량분석법의 생물 탐지체계 적용)

  • Kim, Ju-Hyun
    • Journal of the Korea Institute of Military Science and Technology
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    • v.14 no.2
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    • pp.289-298
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    • 2011
  • TMAH-based Py-MS has been investigated to apply for a real-time classification of biological agents in the field. Acquiring reproducible data from mass spectrometry is a key to biological detection in the field. Nevertheless, it has been little studied on what factors could affect to the reproducibility of the TMAH-based Py-MS spectrum patterns. Given the TMAH-based Py-MS applied to the field system, several factors which could affect to the reproducible pattern of TMAH-based Py-MS spectra are needed to be examined, including changes in TMAH injection volume, growth temperature for microorganism, and number of cells collected in pyrolyzer, and implication of stabilizer used for lyophilization. This study showed that the reproducibility of the spectrum patterns was significantly hindered by changes in TMAH concentration and cell number, and stabilizer implication but not by growth temperature. Among those at low TMAH concentration(0.015m) was not observed the significant alterations of the spectrum pattern even when its injection volume was changed, yet was in different cell numbers and stabilizer implication.

Isolation and Characterization of Acidic Polysaccharides Activating Complement System from the Hot Water Extracts of Pteridium aquilinum var. latiusculum (고사리 열수 추출물로부터 보체계 활성화 산성 다당의 분리 및 특성)

  • O, Byeong-Mi;Gwon, Mi-Hyang;Na, Gyeong-Su
    • The Korean Journal of Food And Nutrition
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    • v.7 no.3
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    • pp.159-168
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    • 1994
  • From the hot water extract of bracken(Pteridium aquilinum var. latiusculum), a Korean win edible plant, anti-complementary acidic polysaccharides were Isolated. Crude polysaccharide fraction(HPA-1) was obtain ed by methanol reflux, ethanol precipitation, dialysis, and lyophilization. HPA-1 contained 81.80% of total sugar, 30.40% of uronic acid, and 15.60cA of protein. HPA 1 was purified consecutively by cetavlon fractionation and chromatography including ion exchange nth DEAE-Sepharose CL 6B and gel permeation with Sephadex G-100 and Sepharose CL-6B. HPA-2- IVa and HPA-Va-2 were nearly homogeneous on HPLC and had 500,000 and 560,000 daltons of molecular weights, respectively. HPA-2-Wa consisted of fucose, galacturonic acid, and glucuronic acid at the molar ratio of 1.40 : 0.97 : 1.88. HPA-2-Va 2 was composed of rhamnose, galactose, and galacturonic acid at the molar ratio of 1.00 : 1.38 : 1.39. The polysaccharides were found to activate the C3 component both In the presence and In the absence of Ca2+ through the crossed-immunoelectrophoresis suggesting that those Involved in both classical and alternative complement pathway.

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Studies on Enzymes of the Higher Fungi of Korea(I) -Identification of Protease in Sarcodon aspratus (한국산(韓國産) 고등(高等) 균류(菌類)의 효소(酵素)에 관한 연구(硏究)(I) -능이의 단백질(蛋白質) 분해효소(分解酵素)의 확인(確認))

  • Park, Wan-Hee
    • The Korean Journal of Mycology
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    • v.14 no.1
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    • pp.25-30
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    • 1986
  • The purposes of this study were to investigate enzyme components and its physiological activities of Sarcodon aspratus (Berk.) S. Ito which grows wildly in Korea, belonging to the family Thelephoraceae. The carpophores of the fungus was extracted with cooling distilled water and salted out by ammonium sulfate. The precipitate was purified by dialysing through visking tube against distilled water and then dissolved with pH 7.8 ammonia aqua, and the extract was filterated. The fraction of filtrate was obtained as light brown powder after lyophilization and determined proteolytic activity. Protease activity of Sarcodon aspratus (Berk.) S. Ito was about two-third of that of pepsin on casein by cup method. The proteolytic potency of this enzyme was found to be 500 unit/mg. This proved the efficacy of the mushroom when it was used as a folk medicine for treating indigestion of beef.

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Effect of Lyoprotectant on the Solubility and Structure of Silk Sericin

  • Kim, Moo-Kon;Kwak, Hyo-Won;Lee, Jeong-Yun;Yun, Hae-Sung;Kim, Min-Hwa;Lee, Ki-Hoon
    • International Journal of Industrial Entomology and Biomaterials
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    • v.25 no.1
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    • pp.133-137
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    • 2012
  • To increase the solubility of lyophilized sericin, we added three types of lyoprotectant: sucrose, trehalose and dextran. The addition of lyoprotectant increased the solubility of lyophilized sericin especially when 1.0% of sucrose, 1.0% and 1.5% trehalose are added. The secondary structure of lyophilized sericin showed that the content of ${\beta}$-sheet or aggregated structure reduced in the presence of lyoprotectant. The morphology of lyophilized was also affected by the addition of lyoprotectant. Whereas flake structure was obtained in the case of pure sericin, a scattered and relatively small flake structure was formed in the presence of lyoprotectant. These finding shows that the presence of lyoprotectant prevents aggregation of sericin molecules and increases the solubility of lyophilized sericin.

Effect of Phaleria nisidai Extracts on Cell Viability of Pancreatic Beta Cell Lines

  • Kim, Seong-Sun;Jeon, Yong-Deon;Kitalong, Christopher;Jin, Jong-Sik
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2019.10a
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    • pp.106-106
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    • 2019
  • The leaves of Phaleria nisidai Kaneh. have been used as a beverage for tonic and immune-enhancing effect in Palau. Recently, extract of the plants showed anti-diabetic effect in clinical trial and possibility could be used as a medicine for diabetes. However, in vitro and in vivo experiments which elucidate cytotoxicity and biological mechanism have not been studied yet. To begin the series of in vitro and in vivo anti-diabetic studies, we evaluated cell viability of pancreatic beta cell lines to figure it out toxicity of the extract. The leaves of Phaleria nisidai were extracted with distilled water and 70% EtOH, and followed by evaporation and lyophilization. And, MIN6 and RIN-m5F cell lines were used for assessment of cell viability. MTT assay was adopted to evaluated a toxicity. Any of extract and concentration used in this study did not show cytotoxicity in the cell lines. Furthermore, morphological change was not observed. These results means Phaleria nisidai might be a medicinal resource when further biological and safety studies would be performed.

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The biomechanical and biological effect of supercooling on cortical bone allograft

  • MuYoung Kim ;Hun-Young Yoon
    • Journal of Veterinary Science
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    • v.24 no.6
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    • pp.79.1-79.16
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    • 2023
  • Background: The need for a storage method capable of preserving the intrinsic properties of bones without using toxic substances has always been raised. Supercooling is a relatively recently introduced preservation method that meets this need. Supercooling refers to the phenomenon of liquid in which the temperature drops below its freezing point without solidifying or crystallizing. Objectives: The purpose of this study was to identify the preservation efficiency and applicability of the supercooling technique as a cortical bone allograft storage modality. Methods: The biomechanical effects of various storage methods, including deep freezing, cryopreservation, lyophilization, glycerol preservation, and supercooling, were evaluated with the three-point banding test, axial compression test, and electron microscopy. Additionally, cortical bone allografts were applied to the radial bone defect in New Zealand White rabbits to determine the biological effects. The degree of bone union was assessed with postoperative clinical signs, radiography, micro-computed tomography, and biomechanical analysis. Results: The biomechanical properties of cortical bone grafts preserved using glycerol and supercooling method were found to be comparable to those of normal bone while also significantly stronger than deep-frozen, cryopreserved, and lyophilized bone grafts. Preclinical research performed in rabbit radial defect models revealed that supercooled and glycerol-preserved bone allografts exhibited significantly better bone union than other groups. Conclusions: Considering the biomechanical and biological superiority, the supercooling technique could be one of the optimal preservation methods for cortical bone allografts. This study will form the basis for a novel application of supercooling as a bone material preservation technique.

The Effects of the Anti-inflammation and Auto-immune System in Mouse by Danchisoyo-san (단치소요산이 염증상태의 면역세포에 미치는 영향)

  • Han Jhee Wan;Han Seong Nim;Ko Soong Gyu
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.16 no.5
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    • pp.914-920
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    • 2002
  • This experiment is carried out for test whether the addition temperament drugs of Danchisoyo-san have an anti-inflammatory effect and have suppression effect on immunocyte in the state of inflammation which induced by carageenan and zymosan. The freezing lyophilization powder which were extracted from Danchisoyo-san divided low dose group(200mg/kg-DSL) and high dose group(500mg/kg-DSH) and after melting in water, it was orally administered to the mouse. Compared with inflammation induced group which were induced by triggering-inflammation reagent carageenan and zymosan and normal contrast group, we measured the edema decrement effect, macrophage and spleen cell activation. 1. Danchisoyo-san has suppress inflammatory reaction induced by carrageenan. 2. Danchisoyo-san has suppress increasing activation of abdominal cavity macrophage cell in the carrageenan induced inflammation. 3. Danchisoyo-san has suppress increasing activation of spleen cell in the carrageenan induced inflammation. 4. Danchisoyo-san has suppress increasing activation of abdominal cavity macrophage in the zymosan induced inflammation. 5. Danchisoyo-san has suppress increasing activation of spleen cell in the zymosan induced inflammation. Based on the above result, Danchisoyo-san was improved its suppression effect to the inflammatory reaction through the suppression of spleen cell and macrophage activation. So we concluded that Danchisoyo-san is prospected as a anti-inflammatory agent to cure inflammation.