2011-08-012024-05-13https://scholars.lib.ntu.edu.tw/handle/123456789/644207摘要：許多文獻中指出，紅血球生成素在對於心臟缺血之後有保護的效果。經由這樣的保護， 可以使缺血後的器官更能夠承受缺血的傷害。對於心肌梗塞病患，如果使用紅血球生成素治療，可以減少心肌受損的程度。在治療心肌梗塞導致心肌壞死的治療中，間葉幹細胞（Mesenchymal stem cells,MSCs）於動物實驗中直接注射於心肌壞死處，可以提高心室收縮能力，甚至分化為心肌細胞幫助壞死心肌的重建。本研究團隊，於前期之研究，發現多醣類高分子，如：幾丁聚醣…等，對大鼠之間葉幹細胞的生長及它再受5-AZA 引導成心肌細的基因表現具有優異性(其結果遠高於一般的細胞培養皿)。因此本研究中擬以紅血球生成素、幾丁聚醣及蠶絲蛋白改質纖維蛋白做為細胞傳遞載體承載大鼠間葉幹細胞作為細胞療法的基礎。此一新材料，可增加生醫材料與組織之黏附性及機械強度並增加其生物活性，並可增進間葉幹細胞於壞死心肌處幫助心肌之重建及血管新生。本研究將探討間葉幹細胞於不同改質纖維蛋白凝膠中生長及分化的情形，促使間葉幹細胞可分化為心肌細胞並促進血管新生。於動物實驗中，將經不同改質之纖維蛋白凝膠包埋分化或未分化之間葉幹細胞以簡單的手術方式塗抹於心肌壞死處，觀察其心臟功能的恢復及並發展新的治療心肌梗塞的方法。同時，紅血球生成素對於間葉幹細胞分化成心肌細胞的幫助機制尚不明確，我們也要藉由此際化觀察紅血球生成素對於間葉幹細胞分化成心肌細胞的影響。本研究將分二部分來進行，第一部份為材料特性研究，主要作法為製備含紅血球生成素、蠶絲蛋白及幾丁聚醣改質纖維蛋白凝膠，探討(1)加入不同濃度之紅血球生成素對纖維蛋白成膠條件、黏度、貼附於組織的能力、降解程度；(2)細胞相容性方向，探討間質幹細胞及血管內皮細胞(共培養下)在紅血球生成素之生物活性、生長及分化的情形(包含MTT，細胞型態染色，fibronectin；GATA4…等RT-PCR 之基因表現)。第二部分：體內實驗，將心肌壞死鼠於壞死位置植入不同成分含有間葉幹細胞之凝膠，探討其不同改質之纖維蛋白凝膠及包覆是否分化之間葉幹細胞對於壞死心肌恢復其功能包含，心輸出量，心臟體積，組織切片分析…等。<br> Abstract: Erythropoietin has protective effects on infarcted myocardium, and through this, the myocardium function could be preserved after infarction.Mesenchymal stem cells (MSCs) cell transplantation is most likely to be effective if it can both: (1) provide a renewable source of proliferating, functional cardiomyocytes (CMCs) and (2) contribute to the development of a network of blood vessels to support and nourish these newly formed cardiomyocytes and the surrounding, ischemic myocardium.We want to combine the two beneficial effects (erythropoietin with MSC), and wish this two beneficial effects could synergic on myocardium protectionIn our previous study, we demonstrated that the new silk fibroin (SF) /polysaccharides (ex: chitosan and hyaluronic acid) hybrid patches may regenerate cardiac infracted tissues. We used the new patches to culture and promote rMSCs proliferation and differentiation to cardiomyogenic cells. The proliferations of the rMSCs on hybrid patches were then quantified. In this investigation, we will develop a new material that use erythropoietin and modified erythopoietin gel to enhance SF gel mechanic strength and adhesion in tissue. MSCs proliferation and differentiation to cardiomyocyte will be induced by 5aza on the patches. We will obtain the effects of the differentiation or undifferentiation of rMSCs in erythropoietin hybrid gel and implanted to rat myocardial.In addition, the effect of erythropoietin on MSC differentiation is still unclear. We will observe the influence of erythropoietin on MSC differentiation to cardiomyocyte.The first year, we will study: (a) fabricating erythropoietin-based hydrogel using different technique to control viscosity…etc; (b) modifying the erythropoietin-based hydrogel with HA, CS and SF or other bioactive molecular; (c) in vitro inducing MSC differentiation into cardiomyocytes in hydrogel. The second year, we will study regeneration of myocardialcytes on a MI rat model by implanting erythropoietin-based hydrogel of different MSC states into the damaged myocardium.