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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 楊台鴻 | |
dc.contributor.author | Hung-Jen Shao | en |
dc.contributor.author | 邵宏仁 | zh_TW |
dc.date.accessioned | 2021-06-15T05:12:07Z | - |
dc.date.available | 2011-07-30 | |
dc.date.copyright | 2010-07-30 | |
dc.date.issued | 2010 | |
dc.date.submitted | 2010-07-23 | |
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dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/46496 | - |
dc.description.abstract | The purpose of this study was to search for a suitable material for the tissue engineering of anterior cruciate ligaments (ACLs) through studying the cell behaviors of ACL cells on materials. Two biodegradable material, chitosan and polycaprolactone (PCL), were included in the first part. We not only evaluated the biocompatibility of ACL cells on these materials but also the potentiality of ACL cells synthesized extracellular matrix (ECM) on these materials. The results indicated ACL cells on chitosan expressed higher level of transforming growth factor β (TGF β) and collagen type III gene than that of ACL cells did on PCL. However, ACL cells presented round cells shape and did not proliferate well on chitosan. In order to overcome this drawback, in the second part, we introduced PCL into chitosan to create chitosan/PCL blended materials and we expected PCL could promote cells to adhere on the materials. The results showed the number of cells adhering on the blended material increased as the PCL content increased in the blends and the behaviors of gene expression of ACL cells were closely related to the cell shape. Therefore, we found using blended materials could make cells be a beneficial shape for proliferating and synthesizing ECM. In the third part, the chitosan/PCL blends were applied to the different cell types. In the study of human bone marrow mesenchymal stem cells (hBMSCs), the results revealed chitosan/PCL blends could induce hBMSCs into chondrogenesis through the alteration of cell shape. In the study of human keratinocyte cell line (HaCaT) and human fibroblast cell line (Hs68) co-culture system, the results illustrated chitosan/PCL blends affected the cell distribution by controlling cells to present different adherent behaviors on the material.
According to our studies, we propose a new concept for the biomaterial selection of tissue engineering. Using blended material could simultaneously control cells toachieve proliferation and phenotypic function. | en |
dc.description.provenance | Made available in DSpace on 2021-06-15T05:12:07Z (GMT). No. of bitstreams: 1 ntu-99-D94548007-1.pdf: 3953854 bytes, checksum: f83c073ff937256af013e214479c6d47 (MD5) Previous issue date: 2010 | en |
dc.description.tableofcontents | 中文摘要 I
Abstract III Content V List of Figures VIII Chapter 1 Introduction 1 1.1 Background 1 1.2 Tissue engineering 2 1.3 Anterior cruciate ligament 2 1.3.1 Histology 3 1.3.2 Ligament cell morphology 4 1.4 Biomaterials 4 1.4.1 Polycaprolactone and chitosan 5 1.4.2 Polycaprolactone and chitosan blends 5 1.5 Interaction of materials and Cell 6 1.6 Growth factors 6 1.7 Experiment design 8 1.8 Flowchart of experiment 9 Chapter 2 Materials and Methods 10 2.1 Materials 10 2.2 Experimental apparatus 13 2.3 Solution preparation 13 2.3.1 Phosphate buffered saline (PBS) 13 2.3.2 MTT reagents 14 2.3.3 2% paraformaldehyde/0.1% Triton X-100 14 2.3.4 Trypsin 14 2.3.5 Western Blot 15 2.4 Preparation of membranes 16 2.5 Isolation and identification of cells 17 2.5.1 Human anterior cruciate ligament cells 17 2.5.2 Human bone marrow mesenchymal stem cells 17 2.6 Cell morphology 18 2.7 Evaluation of cell viability (MTT) 18 2.8 Safranin O staining 19 2.9 Real-time quantitative polymerase chain reaction (PCR) 19 2.10 Western blot analysis 20 2.11 Immunofluorescence study 21 2.12 Histological examinations 22 2.13 Quantification of protein synthesis (ELISA kit) 22 2.14 Flow cytometry 22 2.15 Statistical analysis 23 Chapter 3 Results 24 3.1 Part I: The phenotypic responses of human anterior cruciate ligament cells cultured on PCL and chitosan 24 3.1.1 Adhesion and proliferation of human ACL cells on PCL and chitosan substrates 24 3.1.2 Cytoskeletal structure of human ACL cells on PCL and chitosan substrates 25 3.1.3 Integrin binding analysis of human ACL cells on PCL and chitosan substrates 25 3.1.4 Quantification of FN adsorbed on PCL and chitosan substrates 26 3.1.5 Quantification of FN produced by human ACL cells on PCL and chitosan substrates 26 3.1.6 mRNA expression of human ACL cells on PCL, chitosan and FN-coated chitosan substrates 26 3.1.7 TGF β1 protein expression of Human ACL cells on PCL and chitosan substrates......................................................................................................28 3.2 Part II: Modulation of gene expression and collagen production of anterior cruciate ligament cells through cell shape changes on polycaprolactone/chitosan blends ................................................................29 3.2.1 The FTIR spectra of the blends...........................................................29 3.2.2 Cell morphology and cytoskeletal structure of human ACL cells on the blends ...........................................................................................................29 3.2.3 Proliferation of human ACL cells on the blends.................................30 3.2.4 mRNA expression of human ACL cells on the blends........................30 3.2.5 Human ACL cells treated with cytochalasin C on the blends.............31 3.2.6 Quantification of protein produced by human ACL cells on the blends ......................................................................................................................32 3.3 Part III: chitosan/PCL blends application on different cell types ..................33 3.3.1 Human bone marrow mesenchymal stem cells...................................33 3.3.2 HaCaT (human skin keratinocyte) and Hs68 (human foreskin fibroblasts) ...................................................................................................37 Chapter 4 Discussions..................................................................................................39 4.1 Part I: The phenotypic responses of human anterior cruciate ligament cells cultured on polycaprolactone and chitosan..........................................................39 4.2 Part II: Modulation of gene expression and collagen production of anterior cruciate ligament cells through cell shape changes on polycaprolactone/chitosan blends ...................................................................................................................42 4.3 Part III: chitosan/PCL blends application on different cell types ..................45 4.3.1 Human bone marrow mesenchymal cells ...........................................45 Chapter 5 Conclusion and Perspectives.......................................................................48 Reference .....................................................................................................................50 Appendix......................................................................................................................89 | |
dc.language.iso | en | |
dc.title | 細胞形狀於幾丁聚醣/聚己內酯混摻材料之改變:調控基因改變及細胞分化 | zh_TW |
dc.title | The Alteration of Cell Shape on Chitosan/polycaprolactone Blends: Modulation of Gene Expression and Cell Differentiation | en |
dc.type | Thesis | |
dc.date.schoolyear | 98-2 | |
dc.description.degree | 博士 | |
dc.contributor.coadvisor | 王至弘 | |
dc.contributor.oralexamcommittee | 賴君義,陳志平,宋信文,王淑芬,林克忠 | |
dc.subject.keyword | 前十字韌帶細胞,細胞形狀,基因表現,混摻材料,幾丁聚醣,聚己內酯, | zh_TW |
dc.subject.keyword | anterior cruciate ligament,cell shape,gene expression,blends,chitosan,polycaprolactone, | en |
dc.relation.page | 100 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2010-07-23 | |
dc.contributor.author-college | 工學院 | zh_TW |
dc.contributor.author-dept | 醫學工程學研究所 | zh_TW |
顯示於系所單位: | 醫學工程學研究所 |
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