仿生心臟晶片之壓電系統製程開發及適體分子檢測晶片之研發

Abstract

儘管現今疾病醫療如此進步，藥物的篩檢仍是一費時且漫長的工程，為了提升心臟毒性藥物篩檢之效率，開發一種以壓電為基礎之體外仿生心臟晶片為本研究之主要構想，利用壓電的力電轉換特性電特性，做為即時檢測晶片之優勢，將可應用於動物臨床測試中之快速藥物篩檢，以大幅提升篩檢速率。本研究以壓電高分子聚合物(PVDF、P(VDF-TrFE))之柔性壓電材料做為此晶片薄膜研發之材料，以三維立體結構之微懸臂樑做為其薄膜之結構開發設計，並依心肌細胞之尺度及培養環境需求，研發出低成本壓電薄膜之微機電製程技術。 此外，隨著疾病預防觀念之普及，各類型之檢測需求及其檢測之精準度大幅增加。近年來研究發現，適體(Aptamer)可與大部份分子形成專一性結合之特性、並具有相當之穩定性、以及低成本之可大量製造之優勢，足以做為分子檢測之應用。本研究利用膠體電泳之電泳遷移率分析(EMSA)證實其適體做為分子檢測方法之可行性，且透過自製晶片之設計與機電整合技術研發一微電泳膠體系統，配合即時定量聚合酶鏈鎖反應(Quantitative real time polymerase chain reaction， qPCR)，進一步證實其自製分子檢測晶片之微電泳膠體系統以達到可定性分子之晶片檢測能力，在某程度上具備有一個次級之定量解析能力，顯示其適體檢測方法與晶片未來之可定量分析潛能。

Despite the advances in medical diseases today, drugs screening is still a time-consuming and lengthy process. In order to enhance the efficiency of drug screening for cardiotoxin detection, developing a biomimetic heart on a chip based on piezoelectricity is the major goal of this study. The advantage of using piezoelectric material is its electro-mechanical coupling effect that can provide a real-time signal for monitoring cardiac activities. Furthermore, having an automatic in vitro biomimetic heart on chip system could potentially enhance the drugs screening process during the drug development. In this study, a piezoelectric polymer (PVDF or P(VDF-TrFE)) is used to serve as the flexible structure of the chip. To fit the culture of a single or several myocardial cells, a low cost microfabrication process of a piezoelectric thin-film was developed in this study. Developed process and experimental results will be presented in this thesis. On the other hand, a new methods based-on Aptamer for molecules detection is developed in this srudy. It provides excellent specificity for a large populations of molecules. Further, the production of aptamer is low-cost and stable. Researches have shown that aptamer is a good candidate for future molecular detection methods. In this study, we apply electrophoretic mobility electrophoresis analysis (EMSA) to demonstrate the feasibility of using aptamer for molecular detection. A miniature electrophoresis device was developed for verification of this concept. The real-time quantitative polymerase chain reaction (qPCR) is used to analyze the quantity of detected molecules. It was confirmed that using aptamer to detect molecular is feasible and could potentially provide quantified information.