臺灣博碩士論文加值系統

目前在心血管的研究以及藥物篩檢中，主要藉由心肌細胞的收縮表現、跳動頻率，以及其施力曲線等特徵行為作為檢驗的參數之一，發展出心肌細胞行為檢測平台，而其中以心肌細胞收縮表現的平台，並輔以微機電製程作為主軸，發展柔性結構的設計與微流體系統的設計應用最為廣泛。
然而，在目前的研究中，心肌細胞收縮表現的檢測平台，主要藉由光學儀器觀測細胞本身或柔性基材的形變量，並輔以高解析度的影像，在進行後端處理而得到形變量曲線，間接地得到細胞的施力行為，此即最為普遍得到細胞施力曲線的量測方式。為了能自動化且直接地量測到心肌細胞的施力曲線，本研究提出一種檢測平台的設計，以聚偏氟乙烯共聚合物P(VDF-TrFE) 的柔性壓電聚合材料做為壓電微奈米纖維束換能器的核心，為了使其從粉末狀到成形固化成為微奈米纖維束後可具有壓電特性，本研究開發出以壓電紡絲的製造技術，以快速地製作出多組平行陣列的壓電換能器，並結合智能結構，並同時開發1) 細胞對於壓電基材的從屬性、2) 促使細胞對壓電基材產生依附性及3) 微組織培養技術的三項仿生技術，並成功地誘導心肌微組織產生自發性的跳動。最後，本研究架設可自動化量測心肌微組織自我收縮之介面系統，成功的量測到心肌細胞的週期性收縮訊號，並能夠在投予心臟藥物後，量測到受藥物影響後改變的心臟跳動頻率，實現可自動化且直接性檢測心肌細胞施力訊號及初步藥物篩檢的目標。

In current development for cardiovascular drug discovery, the main parameters are the cardiac systolic and diastolic profiles, beating frequency, and contractile profile. Among these parameters, cardiac systolic and diastolic profiles are the most common, and the monitoring systems are usually based on flexible substrates fabricated by MEMS based microfluidic system is widespread. However, the systems for monitoring cardiac systolic and diastolic profiles are mostly based on an optical systems, and the force profiles is estimated from calculation of the deformation of cells or flexible substrates. Hence, the detection is not direct and could not directly infer relationship between cardiac contraction and drug.
To achieve a fully automatic, real-time and direct massive cardiac drug monitoring system, a platform for real-time monitoring cardiac contractile profile was developed in this study. A piezoelectric material, [poly[(vinylidenefluoride-co-trifluoroethylene]; P(VDF-TrFE:75/25)], was chosen to be the core of the transducer. It is composed of multiple nanofibers to create piezoelectric nanofiber bundles. In order to rapidly develop nanofiber bundles, the electrospinning method was applied. The parallelly oriented piezoelectric nanofiber bundles could be massively fabricated. The overall platforms could be fabricated in one day. A biomimetic substrate coating for facilitating cardiomyocyte adhesion and maturation was also developed. Furthermore, an interface system for monitoring contraction of cardiac micro-tissue was developed and could directly and automatically convert the mechanical force of cardiomyocyte to electrical signals. Also, we verified that this platform can detect cardiac contractile profile by administrating Isoproterenol and Verapamil compounds.

Acknowledgement I
摘要 III
Abstract IV
List of Figures VIII
List of Table XV
Chapter 1 Introduction 1
1.1 Background and Current Limitations of Drug Screening 3
1.2 Current Needs for Drug Screening 5
Chapter 2 Motivation and Purpose 6
Chapter 3 Literature Review 9
3.1 Electrospinning 9
3.1.1 Electrospinning Materials 11
3.1.2 The Patterned Collectors and Spinnerets for Electrospinning 16
3.1.3 The Influence of Environment on Electrospinning 22
3.1.4 The Applications of Electrospun Fibers on Tissue Engineering 25
3.2 Cardiomyocyte Microplate Platform for Drug Screening 28
3.2.1 The Electromechanical Properties of Cardiomyocyte 28
3.2.2 The Methods for Inducing Maturation of Cardiomyocyte 30
3.2.2.1 Mechanical Stimulation 30
3.2.2.2 Electrical Stimulation 33
3.2.2.3 Geometrical Inducement 39
Chapter 4 Development of Nanofiber Bundle with a Controlled Sparation 42
4.1 Material and Method 42
4.1.1 Simulation Parameters 43
4.1.2 Fabrications and Experimental Setup for Electrospinning 45
4.1.3 Sensitivity of Piezoelectric Transducer 48
4.2 Simulation and Experimental Results 49
4.2.1 Dependency of Tooth Angle 49
4.2.2 Dependency of Sharpness 53
4.2.3 Influence of Processing Time 55
4.2.4 Dependency of Pitch Distance 56
4.2.5 Dependency of Gap Distance 60
4.2.6 The Improvement for Defects in The Collector 62
4.2.7 The Sensitivity of Piezoelectric Electrospun Fibers 64
4.2.8 SEM Micrographs of Electrospun Fiber Bundles 65
Chapter 5 Cardiomyocyte Experiment 71
5.1 Material and Method 71
5.1.1 Culture Well Fabrication 71
5.1.2 Device Preparation for Culturing Cardiomyocytes 73
5.1.3 The Cardiomyocyte Culturing 74
5.1.4 The Measurement of Cell Signals 74
5.1.5 Cell Signals without Drug Using 75
5.1.6 Process of Cell Fixing 75
5.1.7 Electrophysiological Study 76
5.1.8 Process of Cell Fixing 76
5.1.9 Immunostaining 77
5.1.10 Microscope for Cell Imaging 78
5.2 Experimental Results 79
5.2.1 Cellular Signals on Substrate with/without Parylene-C Coating 79
5.2.2 Cellular Signals on Substrate with different Thickness of Titanium and SiO2
Coating 80
5.2.3 Contractile Profile of Cultured Cardiomyocytes 81
5.2.4 Cellular Signals after Treating with Isoproterenol and Verapamil 82
5.2.5 Cellular Signals after Electrical Stimulation 91
5.2.6 Time Sequence of Beating Cardiac Cells 92
5.2.7 Immunostaining Results of Cardiac Cells 94
Chapter 6 Conclusions and Future Works 95
6.1 Conclusions 95
6.2 Future Works 96
Reference 97

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