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합성 세포외기질로서의 갈락토실화 키토산에 의한 간실질세포의 형태학적 조절
박인규 ( Park In Gyu ),정환정 ( Jeong Hwan Jeong ),김은미 ( Kim Eun Mi ),범희승 ( Beom Hui Seung ),김수일 ( Kim Su Il ),조종수 ( Jo Jong Su ) 한국키틴키토산학회 2003 한국키틴키토산학회지 Vol.8 No.4
합성 세포외기질을 제조하기 위하여 간실질세포 부착을 위한 갈락토스 잔기를 키토산에 공유결합하였다. 갈락토실화 키토산으로 코팅된 폴리스티렌 디쉬에 대한 간실질세포의 부착은 2시간 경과후에는 94.7%까지 증가한 반면에 대조구로 사용한 키토산으로 코팅한 폴리스티렌 디쉬는 69.1%의 간실질세포 접착율을 보였다. 이는 갈락토실화 키토산 분자와 간실질세포의 아시알로글리코단백철 수용체의 갈락토스 특이적 결합에 의한 것으로 생각된다. 또한 간실질세포의 형태는 갈락토실화 키토산의 농도, 배양시간, 상피성장인자(EGF)에 의해 조절되는 것으로 나타났다. Galactose moieties as the hepatocyte anchorage was covalently coupled with chitosan for the development of synthetic extracellular matrix. Hepatocytes adhesion to galactosylated chitosan (GC)-coated polystyrene (PS) dish became as high as 94.7% after 2h incubation whereas the hepatocytes adhesion to chitosan-coated PS dish was 69.1%, indication of galactosespecific recognition between GC molentration of GC, incubation time, and epidermal growth factor.
Galactosylated chitosan-graft-Poly(ethylene glycol) as hepatocyte-targeting DNA carrier
Cho, Chong-Su 서울대학교 농업개발연구소 2001 농업생명과학연구 Vol.5 No.-
Lactobionic acid bearing galactose group was coupled with chitosan for liver specificity, and poly(ethylene glycol) (PEG) was grafted to galactosylated dchitosan (GC) for stability in water and enhanced cell permeability. Complex formation of galactosylated chitosan-graft-PEG (GCP)/DNA complexes was confirmed by agarose gel electrophoresis. Compared to GC/DNA complex, the stability of GCP/DNA complex could be enhanced. Particle sizes of GCP/DNA complexes decreased as the charge ratio of GCP to DNA increased and had a minimum value around 27 ㎚ at the charge ratio of 5. Conformational change of DNA did not occur after complex formation with GCP compared to conformation of DNA itself. GCP/DNA complexes were only transfected into Hep G2 having asialoglycoprotein receptors (ASGR), indicative of specific interaction of ASGR on cells and galactose ligands on GCP.
강호림,Seog-Jin Seo,Takashi Hoshiba,Toshihiro Akaike,조종수 한국생체재료학회 2008 생체재료학회지 Vol.12 No.3
Galactose moiety was covalently coupled with chitosan to improve hepatocyte adhesion and heparin was immobilized to the galactosylated chitosan (GC) to enhancing hepatocyte function. Hepatocyte adhesion to heparin immobilized GC (GCH)-casted polystyrene (PS) dish was higher than that to chitosan-casted PS dish although it was slightly lower than GC-casted PS dish owing to the hydrated GCH-casted PS dish. Spreading of hepatocytes attached onto GCH-casted PS dish was increased with an increase of heparin for the GCH in the presence of hepatocyte growth factor (HGF). Also, DNA synthesis of the hepatocyte adhered to GCH-casted PS dish was increased with an increase of heparin in the GCH. Higher levels of albumin secretion were maintained on the GCH-casted PS dish than chitosan- or GC-casted PS dishes.
핵의학 영상을 이용한 chitosan의 galactosylation 효과에 대한 평가
정환정 ( Hwan Jeong Jeong ),김은미 ( Eun Mi Kim ),박인규 ( In Kyu Park ),조종수 ( Chong Su Cho ),김창근 ( Chang Guhn Kim ),범희승 ( Hee Seung Bom ) 대한핵의학회 2004 핵의학 분자영상 Vol.38 No.3
목적: 약물이나 유전자 전달에 이용되는 생체적합성이 높은 키토산의 간세포 지향성을 위해서 갈락토스를 수식하는 방법이 널리 이용되어지고 있다. 이번 연구에서는 갈락토스 수식 키토산의 간세포 지향성 획득을 평가하는데 있어서 핵의학 영상법의 유용성에 대해 알아보고자 하였다. 대상 및 방법: 키토산에 NHS와 EDC를 이용하여 30㏖%의 lactobionic acid를 결합시켜 갈락토스 수식 키토산을 제조하였다. 얻어진 갈락토스 수식 키토산을 투석 시킨 후 동 Purpose: Chitosan has been studied as a non-viral gene delivery vector, drug delivery carrier, metal chelater, food additive, and radiopharmaceutical, among other things. Recently, galactose-graft chitosan was studied as a non-viral gene and drug delivery
Jiang, Hu-Lin,Kim, You-Kyoung,Lee, Sun-Mi,Park, Mi-Ran,Kim, Eun-Mi,Jin, Yong-Mei,Arote, Rohidas,Jeong, Hwan-Jeong,Song, Soo-Chang,Cho, Myung-Haing,Cho, Chong-Su 대한약학회 2010 Archives of Pharmacal Research Vol.33 No.4
Hydrogels are widely used in drug delivery systems because they can control the release and thereby enhance the efficiency of locally delivered bioactive molecules such as therapeutic drugs, proteins, or genes. For gene delivery, localized release of plasmid DNA or polymer/DNA complexes can transfect cells and produce sustained protein production. We tested the galactosylated chitosan-graft-polyethylenimine (GC-g-PEI)/DNA complexes-loaded poly(organophosphazene) thermosensitive biodegradable hydrogel as a hepatocyte targeting gene delivery system. The poly(organophosphazene) hydrogel loaded with GC-g-PEI/DNA complexes showed low cytotoxicity and higher transfection efficiency than PEI/DNA complexes, as well as good hepatocyte specificity in vitro and in vivo. Our results indicate that poly(organophosphazene) hydrogels loaded with GC-g-PEI/DNA complexes may be a safe and efficient hepatocyte targeting gene delivery system.
In-Kyu Park,Hu-Lin Jiang,Seung-Eun Cook,조명행,Su-Il Kim,정환정,Toshihiro Akaike,조종수 대한약학회 2004 Archives of Pharmacal Research Vol.27 No.12
Galactosylated chitosan-graft-poly(vinyl pyrrolidone) (GCPVP) was synthesized and characterized for hepatocyte-targeting gene carrier. GCPVP itself as well as GCPVP/DNA complex had negligible cytotoxicity regardless of the concentration of GCPVP and the charge ratio, but GCPVP/DNA complex had slightly cytotoxic effect on HepG2 cells only in the case of the higher charge ratio and 20 mM of Ca2+ concentration used. Through the confocal laser scanning microscopy, it is shown that the endocytosis by interaction between galactose ligands of GCPVP and ASGPR of the hepatocytes was the major route of transfection of GCPVP/F-plasmid complexes.
Park, In-Kyu,Jiang, Hu-Lin,Cook, Seung-Eun,Cho, Myung-Haing,Kim, Su-Il,Jeong, Hwan-Jeong,Akaike, Toshihiro,Cho , Chong-Su The Pharmaceutical Society of Korea 2004 Archives of Pharmacal Research Vol.27 No.12
Galactosylated chitosan-graft-poly(vinyl pyrrolidone) (GCPVP) was synthesized and characterized for hepatocyte-targeting gene carrier. GCPVP itself as well as GCPVP/DNA complex had negligible cytotoxicity regardless of the concentration of GCPVP and the charge ratio, but GCPVP/DNA complex had slightly cytotoxic effect on HepG2 cells only in the case of the higher charge ratio and 20 mM of $Ca^{2+}$ concentration used. Through the confocal laser scanning microscopy, it is shown that the endocytosis by interaction between galactose ligands of GCPVP and ASGPR of the hepatocytes was the major route of transfection of GCPVP/F-plasmid complexes.
Polycaprolactone Scaffold Modified with Galactosylated Chitosan for Hepatocyte Culture
Yuan Qiu,Changyou Gao,Zhengwei Mao,Yimu Zhao,Jichuan Zhang,Qi Guo,Zhongru Gou 한국고분자학회 2012 Macromolecular Research Vol.20 No.3
A suitable extracellular matrix that can maintain long-term proliferation and liver-specific functions of hepatocytes is essential for both extracorporeal bioartificial liver (BAL) assist systems and implantable cell-scaffold constructs. In this work, a polycaprolactone (PCL) scaffold was modified with galactosylated chitosan (GC) to achieve better bioactivity and mechanical stability. The PCL scaffold, prepared by a method of gelatin particle leaching,was hydrolyzed to produce carboxylic groups that were utilized to react with the amine groups of the GC, which was synthesized by grafting the galactose to chitosan. Results showed that the content of carboxylic groups was increased initially with the hydrolysis time until 10 min, at which a COOH density of 2.0×10-5 mol/mg scaffold was reached. The incorporated GC amount showed a positive relationship with the COOH density and finally reached a 100 μg/mg scaffold with a hydrolysis time of 15 min. The incorporated GC was rather stable against incubation in the medium, and could significantly enhance the compression strength of the PCL scaffold in a wet state. With the galactose ligands on the surface, the PCL scaffold could be better recognized by hepatocytes, and show better cell viability, spheroid formation and long-term maintenance of liver-specific functions such as albumin secretion.