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재생의학을 위한 세포친화적 생분해성 지지체의 개발

Title
재생의학을 위한 세포친화적 생분해성 지지체의 개발
Other Titles
DEVELOPMENT OF CELL-INTERACTIVE BIODEGRADABLE SCAFFOLDS FOR REGENERATIVE MEDICINE
Author
신영민
Alternative Author(s)
Shin, Young Min
Advisor(s)
신흥수
Issue Date
2011-02
Publisher
한양대학교
Degree
Doctor
Abstract
많은 국가들이 고령화 사회에 접어듦에 따라 사고나 외상으로 인해 손상된 장기/조직 등의 재생 소요가 증가하고, 이를 위해 적절한 지지체를 기반으로 손상된 장기에 세포를 전달하는 재생의학 (또는 조직공학) 이 개발되어 다양한 연구가 진행되고 있다. 흡착, 증식, 분화 등의 일련의 세포반응을 조절할 수 있는 재생의학 지지체의 개발을 위해 생체친화성을 갖는 PLGA, PLLA, PCL 등의 생분해성 합성고분자들이 활발하게 이용되고 있으나, 현재의 지지체 형태 및 기능은 세포외기질 (ECM)과는 큰 차이가 있어 새로운 지지체 개발의 소요가 발생되었으며, 최근 생체모사공학과 접목되어 세포외기질과 구조적/기능적으로 더욱 가까운 지지체의 개발이 가속화되고 있다. 그래서 본 논문에서는 다양한 표면 개질 기술을 이용하여 세포친화적인 생분해성 지지체를 개발하였다. 먼저 표면에 기능기 또는 세포반응기가 없는 합성고분자 지지체는 감마선을 이용하여 카르복실기가 도입되었으며, 이는 생체활성물질인 젤라틴 또는 RGD 펩타이드의 표면 결합에 사용되었다. 젤라틴이 도입된 지지체는 성체줄기세포의 흡착 및 성장을 촉진시켰을 뿐만 아니라 골분화까지 긍정적인 영향을 주었으며, RGD 펩타이드가 고정된 지지체는 골아세포의 흡착, 증식, 분화를 촉진하여 기존에 사용되었던 지지체에 비해 높은 세포친화성을 보여주었다. 또, 새로운 표면 개질 기술로써, 폴리도파민 코팅기술을 이용하여 세포친화적인 지지체를 개발하였다. 폴리도파민은 다양한 코팅조건에서 코팅의 정도가 조절되었으며, 코팅수준에 따른 근아세포 반응성의 차이를 보여주었다. 생체활성물질로써 젤라틴, RGD 펩타이드, 그리고 성장인자가 추가적으로 도입된 지지체를 개발하였는데, 젤라틴이 도입된 지지체는 근아세포의 흡착, 증식을 2배 가량 높여주었다. 또한 RGD 펩타이드가 도입된 지지체는 각각 근아세포와 골아세포의 세포반응성을 매우 높여주어 기능성 심근패치와 골조직 재생용 지지체로써의 기능을 할 수 있을 것으로 생각된다. 조직의 원활한 재생 및 분화에 영향을 주는 요인인 성장인자로써 혈관내피세포 성장인자가 도입된 지지체를 개발하였고, 이는 혈관내피세포의 흡착, 이동, 증식에 매우 긍정적인 영향을 주어 인공혈관에 매우 유용하게 사용될 수 있을 것으로 보인다. 종합적으로 감마선과 폴리도파민 코팅 기술을 이용하여 다양한 조직 재생을 위한 지지체를 개발하였는데, 이들 지지체는 생체활성물질의 추가적 도입을 통해 높은 세포 친화성을 유지하였으며, 세포 특이적인 분화에도 영향을 줌에 따라 다양한 조직 재생을 위해 본 기술이 유용하게 사용될 수 있을 것이다.|As most of countries have been rapidly advancing to the aging society, regenerative medicine (or tissue engineering) has been designed to replace damaged tissues caused by accident and trauma with appropriately delivering cells and/or growth factors using biocompatible materials as carriers. To provide structural platform to cells regulating their survival events such as adhesion, spreading, proliferation, and differentiation, the scaffolds have been developed with biocompatible synthetic materials such as PLGA, PLLA, and PCL. However, the shape or function of scaffolds is far from the properties of native extracellular matrix (ECM), therefore biomimetic approach has been combined to narrow the gap between engineered scaffolds and natural ECM in tissue regenerative application. In this thesis, I developed cell-interactive scaffolds based on synthetic biomaterials without surface functional groups and cell-interactive cues using gamma-ray irradiation, carboxylic acid groups were firstly introduced and subsequently used to conjugate gelatin or RGD peptide as cell-interactive molecules. The gelatin or RGD peptide immobilized scaffolds enhanced adhesion, spreading and proliferation of hMSC and preosteoblastic cells. In addition, gelatin immobilized scaffolds enhanced osteogenic differentiation approximately 2-times greater of than that of non-modified scaffolds, the scaffolds conjugated with RGD peptide also enhanced osteogenic gene expression directing that surface immobilized bioactive molecules can regulate behaviors of cells on the scaffolds. As another surface modification strategy, I used polydopamine coating method to develop cell-interactive scaffolds. I found that generation of polydopamine layer on the scaffolds was regulated with a variety of coating condition, which modulated a series of cellular responses on the scaffolds. Adhesion and proliferation of cardiac myoblasts was approximately 2-times greater than those on the non-modified scaffolds, and even very low amounts of coated polydopamine can successfully regulate cell behaviors. Gelatin, RGD peptide, and growth factors as bioactive molecules, were further immobilized on the polydopamine coated scaffolds, and the immobilized bioactive molecules maintained their bioactivity. Gelatin immobilized scaffolds successfully regulated adhesion, spreading, and proliferation of cardiac myoblasts (approximately 2-time greater than non-modified scaffolds), and RGD peptide immobilized scaffolds also modulated the survival events of cardiac myoblasts and preosteoblastic cells generating functional cardiac patch and bone tissue engineering scaffolds. In addition, I proved that VEGF immobilized on the polydopamine coated scaffolds play a role in mediating HUVEC survival such as adhesion, migration, and proliferation, and another growth factor also regulated the behaviors of the cells. Therefore, the cell-interactive scaffolds may be novel candidates for regeneration of the defected myocardium, bone, and vessels.
As most of countries have been rapidly advancing to the aging society, regenerative medicine (or tissue engineering) has been designed to replace damaged tissues caused by accident and trauma with appropriately delivering cells and/or growth factors using biocompatible materials as carriers. To provide structural platform to cells regulating their survival events such as adhesion, spreading, proliferation, and differentiation, the scaffolds have been developed with biocompatible synthetic materials such as PLGA, PLLA, and PCL. However, the shape or function of scaffolds is far from the properties of native extracellular matrix (ECM), therefore biomimetic approach has been combined to narrow the gap between engineered scaffolds and natural ECM in tissue regenerative application. In this thesis, I developed cell-interactive scaffolds based on synthetic biomaterials without surface functional groups and cell-interactive cues using gamma-ray irradiation, carboxylic acid groups were firstly introduced and subsequently used to conjugate gelatin or RGD peptide as cell-interactive molecules. The gelatin or RGD peptide immobilized scaffolds enhanced adhesion, spreading and proliferation of hMSC and preosteoblastic cells. In addition, gelatin immobilized scaffolds enhanced osteogenic differentiation approximately 2-times greater of than that of non-modified scaffolds, the scaffolds conjugated with RGD peptide also enhanced osteogenic gene expression directing that surface immobilized bioactive molecules can regulate behaviors of cells on the scaffolds. As another surface modification strategy, I used polydopamine coating method to develop cell-interactive scaffolds. I found that generation of polydopamine layer on the scaffolds was regulated with a variety of coating condition, which modulated a series of cellular responses on the scaffolds. Adhesion and proliferation of cardiac myoblasts was approximately 2-times greater than those on the non-modified scaffolds, and even very low amounts of coated polydopamine can successfully regulate cell behaviors. Gelatin, RGD peptide, and growth factors as bioactive molecules, were further immobilized on the polydopamine coated scaffolds, and the immobilized bioactive molecules maintained their bioactivity. Gelatin immobilized scaffolds successfully regulated adhesion, spreading, and proliferation of cardiac myoblasts (approximately 2-time greater than non-modified scaffolds), and RGD peptide immobilized scaffolds also modulated the survival events of cardiac myoblasts and preosteoblastic cells generating functional cardiac patch and bone tissue engineering scaffolds. In addition, I proved that VEGF immobilized on the polydopamine coated scaffolds play a role in mediating HUVEC survival such as adhesion, migration, and proliferation, and another growth factor also regulated the behaviors of the cells. Therefore, the cell-interactive scaffolds may be novel candidates for regeneration of the defected myocardium, bone, and vessels.
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http://dcollection.hanyang.ac.kr/jsp/common/DcLoOrgPer.jsp?sItemId=000000059315https://repository.hanyang.ac.kr/handle/20.500.11754/140019
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GRADUATE SCHOOL[S](대학원) > BIOENGINEERING(생명공학과) > Theses (Ph.D.)
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