2011-08-012024-05-14https://scholars.lib.ntu.edu.tw/handle/123456789/658812摘要：本研究使用電紡的技術將SF 和HA 複合物製作具方向的薄膜接之上紅血球生成素(EPO)並結合此複合物之微粒做為血管內皮細胞生長因子釋放之載體，於其上培養人類羊水幹細胞(hAES)觀察其生長及分化，並進行大動物試驗的體內測試。本研究將SF/HA 等經電紡製作成具方向性之材料表面，使用光化學反應將EPO 接枝於SF/HA patch 將VEGF 吸入SF/HA 複合微小球來進行釋放，hAES 於其上之細胞行為表現及大動物實驗的體內評估。此一新材料預估可增加壞死心肌部分之血管新生，並增進hAES 於壞死心肌處幫助心肌之重建及血管新生。本研究將分三部分來進行，第一部分為材料製備，首先製備SF/HA 能使細胞具方向性生長之電紡薄膜，比較不同強度電壓下、噴嘴頭直徑及不同成分溶劑和溶液黏度所製出的SF/HA 薄膜其機械強度及薄膜表面方向性。第二部份為製備接枝有紅血球生成素之蠶絲蛋白/玻尿酸薄膜，並製作吸附有VEGF 之SF/HA 微小球，探討(1)使用不同濃度之紅血球生成素在SF/HA 之接枝效率、AES 之細胞行為表現；(2)將控制SF/HA 微小球表面膨潤度，探討吸附不同濃度之VEGF 之吸附效率及釋放速率，及對培養其上之hAES 細胞型態及分化造成的影響。第三部分：體內實驗，進行大動物實驗，置SF/HA-EPOpatch 培養hAES，探討其促進分化及血管新生之效率對於壞死心肌恢復其功。<br> Abstract: In our previously studies, the results demonstrate that the novel SF/HA and SF/CS-HAhybrid patches may be promising biomaterials for regenerating infarcted cardiac tissues. Thestudies suggest that CD44 of rMSCs promote the growth of rMSCs, fibronectin expression,and cardiomyogenic differentiation of 5-aza inducing rMSCs on SF/HA patches than SFpatches while the cells with a CD44-blockage treatment reverse those promotions. In animalstudy, we investigated the effects of tissue engineered cardiac patches consisting of SF/HApatches on attenuation of LV dysfunction. The small area of myocardial fibrotic scardeveloped, and the resulting decreased contractility in the corresponding area was notassociated with diastolic dilation. New cardiac SF/HA patches for regenerating cardiactissues have a great potential.Erythropoietin has protective effects on infarcted myocardium, and through this, themyocardium function could be preserved after infarction. Human amniotic epithelial stemcell (AES) is considered as medical wastes after amniocentesis and can be obtained withoutethical controversy; its derived stem cells have been induced to differentiation to cell typesrepresenting each embryonic germ layer: (1) provide a renewable source of proliferating,functional cardiomyocytes (CMCs) and (2) contribute to the development of a network ofblood vessels to support and nourish these newly formed cardiomyocytes and thesurrounding, ischemic myocardium.We want to combine the two beneficial effects (erythropoietin with AES), and wish thistwo beneficial effects could synergic on myocardium protection in our previous study, wedemonstrated that the new silk fibroin (SF) /hyaluronic acid (HA) hybrid patches mayregenerate cardiac infracted tissues. We used isotropic of the new patches to makeelectrospinning and promote hAES proliferation and regeneration heart in vivo. Theproliferations of hAES on SF/HA patches were then quantified.In this investigation, we will develop a new material that use erythropoietin and modifiederythopoietin graft to enhance SF/HA patch mechanic strength and adhesion in tissue. hAESproliferation and differentiation to cardiomyocyte will be induced by on the patches invivo. We will obtain the effects of the differentiation or undifferentiation of hAES inerythropoietin grafted SF/HA patches releasing VEGF and implanted to porcine myocardial.We will be made isotropic SF /HA patch by electrospinning and observe the influence oferythropoietin on hAES differentiation to cardiomyocyte. The first year, we will study: (a)fabricating isotropic SF/HA patch by electrospinning using different technique to controlviscosity…etc; (b) modifying the erythropoietin-based or controlling release VEGF onSF/HA patch; The second and third year, (a) in vitro proliferating hAES into cardiomyocytesin SF/HA patch. We will study regeneration of cardiomyocytes on a MI porcine model byimplanting erythropoietin-grafted on SF/HA patch plus VEGF releasing of hAES states intothe damaged myocardium.