DC 欄位 | 值 | 語言 |
dc.contributor | 林峰輝 | zh-TW |
dc.contributor | 臺灣大學:醫學工程學研究所 | zh-TW |
dc.contributor.author | 莊子文 | zh-TW |
dc.contributor.author | Chuang, Tzu-Wen | en |
dc.creator | 莊子文 | zh-TW |
dc.creator | Chuang, Tzu-Wen | en |
dc.date | 2008 | en |
dc.date.accessioned | 2010-06-02T01:19:37Z | - |
dc.date.accessioned | 2018-06-29T00:48:05Z | - |
dc.date.available | 2010-06-02T01:19:37Z | - |
dc.date.available | 2018-06-29T00:48:05Z | - |
dc.date.issued | 2008 | - |
dc.identifier.other | U0001-3101200816113100 | en |
dc.identifier.uri | http://ntur.lib.ntu.edu.tw//handle/246246/184612 | - |
dc.description.abstract | 冠心病是國人常見的疾病,此疾病是因提供心臟所需的冠狀動脈發生狹窄阻塞所致。目前常見的治療方式是支架的置入,但支架不理想的血液相容性,常使得撐開後的血管在半年後再度的狹窄。為了避免血管的再狹窄,本研究固定抗凝血藥物肝素於支架表面以改善其血液相容性。由於金屬支架表面並不具有具反應性之官能基,本研究先以表面偶合劑二異氰酸己烷將之活化,使支架得以進行後續的藥物固定反應。支架表面改質後,肝素分子也必須進行活化,方可固定於支架表面。乙基二甲基氨丙基碳二亞胺是目前常用來固定肝素至材料表面的交聯劑。此試劑可將肝素的羧酸基轉換成高反應性的中間產物,使肝素固定於支架表面。但有研究指出,經此試劑處理後的肝素,將喪失其部分的抗凝血功效。為了避免羧酸基的耗損,本研究以氧化劑過碘酸鈉處理肝素。此分子可使肝素分子中特定且無關肝素活性的氫氧基轉成極具反應性的醛基,使肝素能在不損失羧酸基的情況下,固定於材料表面。本研究我們分別以乙基二甲基氨丙基碳二亞胺及過碘酸鈉將肝素固定於支架表面,並進行各種肝素活性及血液相容性測試。研究結果顯示,以乙基二甲基氨丙基碳二亞胺法固定肝素後之支架較未經處理之支架有相對較好的血液相容性,但結果仍未盡理想。而經過碘酸鈉法固定肝素之支架,則有明顯較好的血液相容性。因此我們相信,本研究所研發的肝素固定技術,能改善過去支架不理想的血液相容性,解決血管再狹窄的問題。 | zh-TW |
dc.description.abstract | Poor compatibility between blood and metallic coronary artery stents is one reason for arterial restenosis. Immobilization of anticoagulant heparin on the stent’s surface is feasible for improving compatibility. Prior to heparin immobilization, we examined possible surface-coupling agents for heparin immobilization. Hexamethylene diisocyanate (HMDI) and 3-aminopropyl-triethoxysilane (APTS) were examined as surface-coupling agents to activate 316L stainless steel (e.g., stent material). The activated surface was characterized by Fourier transformation infrared spectroscopy (FTIR), atomic force microscope (AFM), surface plasmon resonance (SPR), and trinitrobenzene sulfonic acid (TNBS) assay. In the FTIR analysis, HMDI and APTS were both covalently linked to 316L stainless steel. In the AFM analysis, it was found that the HMDI-activated surface was smoother than the APTS-activated one. In the SPR test, the shift of the SPR angle for the APTS-activated surface was much higher than that for the HMDI-activated surface after being challenged with acidic solution. The TNBS assay was utilized to determine the amount of immobilized primary amine groups. The HMDI-activated surface was found to consist of about 1.32 μmole/cm2 amine group, whereas the APTS-activated surface consisted of only 0.89μmole/cm2 amine group. We conclude that the HMDI-activated surface has more desirable surface characteristics than the APTS-activated surface, such as surface roughness, chemical stability, and the amount of active amine groups. The HMDI-activated 316L stainless steel (SS) was then utilized for heparin linking. The compound 1-ethyl-3-(3-dimethyl-aminopropyl) carbodiimide (EDAC) has often been utilized for the immobilization of heparin, but the critical carboxyl groups of heparin (with regards to heparin’s anticoagulant activity) will be reduced by this method. We were trying to examined possible methods of heparin immobilization without consuming these carboxyl groups. Sodium periodate (NaIO4; SP) was then used to oxidize heparin to form aldehyde groups and then coupled with bis-amine-terminated poly(ethylene glycol) (Bis-amine PEG) so as to form heparin-PEG complexes. The complexe could then be grafted onto the activated surface of the test material without losing its carboxyl groups. The heparin-PEG complex formed by EDAC method was used as comparison group. Effective surface modification of the HMDI-activated and heparin-PEG grafted 316L SS surface was confirmed using Fourier Transform Infrared Spectroscopy (FTIR), Electron Spectroscopy for Chemical Analysis (ESCA) and a water contact angle test. After the heparin grafted by SP, the surface showed an improvement in antithrombrin (AT) binding ability, its anticoagulant property, and hemocompatibility in comparison to heparin grafted by EDAC. | en |
dc.description.tableofcontents | ABSTRACT 1HAPTER 1 INTRODUCTION-1 Coronary atherosclerosis 4-2 Percutaneous transluminal coronary angioplasty 6-3 The pathophysiology of restenosis after angioplasty 8-4 Stent implantation 10-5 Pathology of in-stent restenosis 12-6 Treatment of in-stent restenosis 13-6-1 Brachytherapy 13-6-2 Rotational atherectomy 13-6-3 Cutting balloon 14-6-4 Laser angioplasty 15-6-5 Drug-eluting stents 15-7 Objectives of this study 19HAPTER 2 THEORETICAL BASIS-1 Blood-compatible materials 21-2 Heparinized surface 22-3 316L Stainless Steel 24-4 Electropolish for 316L SS stent surface 25-5 Surface activation for 316L SS 26-6 The methods for heparin immobilization 27HAPTER 3 MATERIALS AND METHODS-1 Surface pretreatment and activation for 316L SS 34-1-1 Electropolishing process 34-1-2 Preparation of HMDI-activated 316L SS 34-1-3 Preparation of APTS-activated 316L SS 35-2 Surface characterization of the activated 316L SS 35-2-1 FTIR spectra for the activated 316L SS 35-2-2 AFM topography of the activated 316L SS 35-2-3 TNBS assay for the activated 316L SS 36-2-4 SPR analysis for the stability of the HMDI and APTS activated surface 36-2-5 Surface immobilizing of PEG 37-3 Preparation and characterization of activated heparin for immobilization 38-3-1 Preparation and toluidine blue testing of EDAC- or SP-modified heparin 38-3-2 FTIR spectra for EDAC- and SP-treated heparin 38-3-3 Purpald assay for the SP-treated heparin 39-4 Preparation and characterization of heparin-PEG complexes 39-4-1-1Preparation of the heparin–PEG complexes with EDAC method 39-4-1-2Preparation of the heparin–PEG complexes with SP method 40-4-2 FTIR spectra for the two heparin–PEG complexes 40-4-3 SPR test for the interaction of grafted AT and the two heparin-PEG complexes 41-4-4 SPR testing for affinity measurement of grafted heparin and AT 41-4-5 Blood coagulation assays of the two heparin-PEG complexes 42-5 Surface immobilization and characterization of the two heparin–PEG complexes 42-5-1 Surface immobilization of the two heparin–PEG complexes 42-5-2 Surface characterization of the heparinized surfaces with water contact ngle test, ESCA, and toluidine blue assay 43-5-3 AT adsorption testing of the heparinized 316L SS 44-5-4 Preparation of the AT-grafted probe and AFM measurement 45-5-5 Hemocompatibility characterization of the heparinized 316L SS 45-6 LDH and WST-1 test for in vitro biocompatibility of native and heparinized 316L SS 46-6-1 LDH leakage 46-6-2 WST-1 Assay 47HAPTER 4 RESULTS-1 Characterization of HMDI and APTS-activated 316L SS 48-1-1 FTIR spectra for the HMDI-activated 316L SS 48-1-2 FTIR spectra for the APTS-activated 316L SS 50-1-3 AFM topography of the activated 316L SS 51-1-4 SPR analysis for the stability of the HMDI and APTS activated surface. 52-1-5 TNBS assay for the activated 316L SS 54-1-6 FTIR spectra for the PEG grafted 316L SS 55-2 Characterization of the EDAC- and SP-treated heparin 57-2-1 FTIR spectra for EDAC heparin 57-2-2 The number of aldehyde groups per heparin molecular versus the SP concentration for the heparin activation 58-2-3 Fourier Transform Infrared spectra for SP-treated heparin 59-2-4 Toluidine blue testing of EDAC- or SP-modified heparin 60-3 Characterization of the two heparin-PEG complexes 61-3-1 FTIR spectra of the bis-amine PEG, the native heparin, the heparin-PEG complex formed by EDAC method, and the heparin-PEG complex ormed by SP method 61-3-2 SPR test for the interaction of grafted AT and the heparin-PEG omplex 63-3-3 SPR test for the interaction of grafted AT and the heparin-PEG 66omplex-3-4 Blood coagulation assays of the heparin-PEG complexes 69-4 Surface characterization of the heparinized surface 70-4-1 Surface characterization of the heparinized surface with water contact 70ngle test-4-2 Surface characterization of heparinized surface with ESCA 72-4-3 Estimation of surface heparin density with toluidine blue assay 73-4-5 AT adsorption testing of the heparinized 316L SS 74-4-6 AFM measurement of the adhesion force of the heparinized surface nd AT grafted probe 75-4-7 Hemocompatibility characterization of the heparinized 316L SS 77-5 LDH and WST-1 test of the heparinized 316L SS 80HAPTER 5 DISCUSSIONS 84HAPTER 6 CONCLUSION 96EFERENCE 98ESUME 111IGURES AND TABLESigure 1-1 Balloon Angioplasty 7igure 1-2 Restenosis 8igure 1-3 Stent implantation 11igure 1-4 Rotational Athrectomy 14igure 1-5 The four steps for heparin immobilization 20igure 2-1 The interaction of grafted heparin with AT 23igure 2-2 Proposed mechanism for reaction of hexamethylene diisocyanate (HMDI) with 316L stainless steel surface. 26igure 2-3 The structure of AT binding sequence of heparin 28igure 2-4 The heparin carboxyl group is consumed during the activation rocess 29igure 2-5 The hydroxyl group can react with cyanogen bromide to give the reactive cyclic imido-carbonate 30igure 2-6 The hydroxyl group can react with chloroformates to give the intermediates that produced by cyanogens bromide 30igure 2-7 DSC can activate hydroxylic compounds to form an amine reactive intermediate 31igure 2-8 CDI can activate hydroxylic compounds to form an amine reactive intermediate 31igure 2-9 The HMDI can activate hydroxylic compounds to form an amine reactive intermediate 32igure 2-10 The succinic anhydride can create terminal carboxyl group on hydroxyl bearing compound in aqueous environment 32igure 4-1 FTIR spectra of the HMDI monomer and HMDI-activated 16L SS 49igure 4-2 FTIR spectra of the APTS monomer and APTS-activated 16L SS 51igure 4-3 Noncontact AFM images of native stainless steel, electropolished stainless steel, HMDI-activated 316L SS, and the APTS-activated 316L SS 52igure 4-4 Surface plasmon resonance angle of the native, the activated, 53nd the activated surface after washing with 0.1NHCl. igure 4-5 Reflected light intensity versus time for the activated surface ith 0.1N HCl washing 54igure 4-6 Amount of amine groups on the APTS-activated and HMDI-ctivated 316L SS 55igure 4-7 FTIR spectra of bis-amine and bis-amine-modified 316L SS 56igure 4-8 FTIR spectra of native heparin, and of native heparin treated ith EDAC 58igure 4-9 The number of aldehyde groups per heparin molecular versus he SP concentration for the heparin activation 59igure 4-10 FTIR spectra of native heparin, and of native heparin treated ith SP. 60igure 4-11 OD value for the toluidine blue solution added to native heparin nd various concentrations of EDAC or SP. 61igure 4-12 FTIR spectra of the bis-amine PEG, the native heparin, the eparin PEG complex formed by EDAC method, and the eparin-PEG complex formed by SP method. 63igure 4-13 The design of the SPR tests 64igure 4-14 The schematic sensorgram showing the results of antithrombin coupling using EDAC 65igure 4-15 The sensorgram showing interaction of grafted AT with native heparin, heparin-PEG complexes formed by SP or EDAC method 66igure 4-16 The schematic sensorgram showing the results of bis-amine PEG coupling using EDAC and the interaction of SP-treated heparin ith bis-amine PEG on the SPR sensing surface. 67igure 4-17 The sensorgram showing the interaction of the SP or EDAC ethod grafted heparin AT 68igure 4-18 Water contact angle on surfaces of the native 316L SS, the HMDI activated 316L SS, the bis-amine PEG-modified 316L SS, the SP heparin-immobilization group, and the EDAC heparin-mmobilization group 71igure 4-19 ESCA survey scans spectra of the control group (native 316L SS), P heparin-immobilization group and EDAC heparin-mmobilization group. 73igure 4-20 The amount of adsorbed AT on the surfaces of the SP heparin immobilization group, EDAC heparin-immobilization group and control group (native 316L SS). 75igure 4-21 The force-versus-distance curve (f-d curve) between the AT rafted AFM probe and the native 316L SS, SP method modified16L SS surface, and EDAC method modified 316L SS surface. 76igure 4-22 Scanning electron micrographs of thrombus adherence to the urface of the control group, the EDAC heparin-immobilization roup, and the SP heparin-immobilization group. 77igure 4-23 Scanning electron micrographs of thrombus adherence to the urface of the control group 78igure 4-24 Scanning electron micrographs of thrombus adherence to the urface of the EDAC heparin-immobilization group. 79igure 4-25 Scanning electron micrographs of thrombus adherence to thesurface of the SP heparin-immobilization group. 80igure 4-26 The LDH test for in vitro biocompatibility of native and eparinized 316L SS (3T3) 81igure 4-27 The LDH test for in vitro biocompatibility of native and eparinized 316L SS (HUVEC cells) 81igure 4-28 The WST-1 test for in vitro biocompatibility of native and eparinized 316L SS (3T3) 82igure 4-29 The WST-1 test for in vitro biocompatibility of native and eparinized 316L SS (HUVEC cells) 83able 4-1 Activated partial thromboplastin time (APTT), prothrombin time PT), and thrombin time (TT) of the native heparin and the two hepairn-PEG complexes. 70able 4-2 The chemical compositions of native 316L SS and 316L SS odified step-by-step with HMDI, BA-PEG and SP-treated or DAC-treated heparin. 73 | en |
dc.format | application/pdf | en |
dc.format.extent | 11336159 bytes | - |
dc.format.mimetype | application/pdf | - |
dc.language | en | en |
dc.language.iso | en_US | - |
dc.subject | 關鍵字: 支架 | zh-TW |
dc.subject | 肝素 | zh-TW |
dc.subject | 表面改質 | zh-TW |
dc.subject | 表面分析 | zh-TW |
dc.subject | 血液相容性 | zh-TW |
dc.subject | Keywords: stent | en |
dc.subject | heparin | en |
dc.subject | surface modification | en |
dc.subject | surface characterization | en |
dc.subject | hemocompatibility | en |
dc.title | 316L不鏽鋼表面活化改質及固定抗凝血藥物肝素於冠心病支架上之應用 | zh-TW |
dc.title | Immobilization of Anticoagulant Heparin on HMDI-activated 316L Stainless Steel for the Application of Coronary Artery Disease | en |
dc.type | thesis | en |
dc.identifier.uri.fulltext | http://ntur.lib.ntu.edu.tw/bitstream/246246/184612/1/ntu-97-F91548001-1.pdf | - |
item.languageiso639-1 | en_US | - |
item.cerifentitytype | Publications | - |
item.fulltext | with fulltext | - |
item.openairecristype | http://purl.org/coar/resource_type/c_46ec | - |
item.openairetype | thesis | - |
item.grantfulltext | open | - |
顯示於: | 醫學工程學研究所
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