2015-08-012024-05-14https://scholars.lib.ntu.edu.tw/handle/123456789/659413摘要：本團隊曾發展出的蠶絲-玻尿酸/骨髓間葉幹細胞（SF-HA/BMSC )心肌綴補片放置於MI大鼠及 豬隻進行心肌修復研究，顯示植入SF-HA/BMSC心肌綴補片可以減低心室擴張及改善心肌功能及增 進受損心肌區域的再生。本研究擬與共同主持人陽明大學醫工系鍾次文教授發展新一代具彈性及生物 活性之PCL-SF (蠶絲)為基礎之心肌綴補片及支架，如：PCL-SF、PCL-SF/ HA 'PCL-SF/HA/ GRGD 等，配合BMSC的心肌分化能力，作為修復受損心肌區域的生醫材料。初步研究結果，發現 PCL-SF/HA/GRGD具有與細胞CD44 receptor、integrin結合之雙功能，此心肌綴補片於體外心肌分 化時，與其他材料比較，可誘導出BMSC細胞團（BMSC micromass )並且有很高之心肌蛋白（如： Connexin 43)的表現量；團隊將以此材料與細胞團為基礎，研究此心肌綴補片/BMSC於體外心肌分化 及動物體内心肌修復的效果。另外，因為BMSC細胞球具有優良的特定細胞的分化效果，本研究擬對 支架/BMSC細胞球也進行與心肌綴補片/BMSC相同的研究。本三年期研究計晝，分為：第一部分， 探討具單或雙功能之PCL-SF/ HA或HA/GRGD心肌綴補片的製備及材料特性分析，如：FT-IR及XPS 對功能性分子之接枝的表面分析；生物活性之研究包含BMSC體外增生及心肌分化的特性，如：以 MTS與Live/Dead檢測細胞增生或形成細胞團之觀察；以共輛聚焦和RT-PCR檢測心肌分化蛋白及 paracrine factors 之基因變化，如：Connexin 43 及 VEGF...等；第二部分，探討 PCL-SF/HA/GRGD 支 架/ BMSC細胞球系統：支架的製備及材料特性分析，如：支架孔隙度及孔徑大小及對功能性分子之 接枝的表面分析；BMSC細胞球製備及生物特性分析，如：球體增生及心肌分化的特性，BMSC細 胞migration特性及PCL-SF/HA/GRGD支架對球體生物特性之影響；第三部分，探討單或雙功能心 肌綴補片/BMSC及支架/ BMSC細胞球系統植入MI鼠心肌8周後，MI鼠的心肌功能恢復情形並藉 由MRI、組織染色...等方式，來觀察心室擴張及心肌功能改善情形及增進受損心肌區域血管的新生， 如同本團隊以前之研究。藉由本研究，新一代具彈性及生物活性之PCL-SF (蠶絲)為基礎之心肌綴補 片及支架於MI大鼠的心肌修復功能及機制再深入了解，可作為臨床轉譯前的重要根據。<br> Abstract: SF-HA/BMSC cardiac patch have been developed for cardiac repair of MI hearts of rats and pigs. The patch show the good efficacies of cardiac repairs including reducing remodeling, improving LV functions, inhibiting apoptosis of cardiomyocytes and enhancing angiogenesis of MI hearts of rats and pigs.Our team would like develop a new elastic and high bioactive PCL-SF based patch and scaffolds incorporated with cardiomyogenesis capacity of BMSC for cardiac repair of MI hearts. The preliminary results showed that grafting dual bio-functional molecules into PCL/SF to produce PCL-SF/HA/GRGD patch induced BMSC micro-mass with higher cardiac proteins expressions (e.g., Connexin 43) compared with others. We will investigate PCL-SF/HA/GRGD/ BMSC patch on the efficacies of in-vitro cardiomyogenesis and in-vivo cardiac repairs in a rat MI heart model. Since it has been reported that BMSC spheroids have superior differentiation properties compared with BMSC cells, the in-vitro and in-vivo studies for PCL-SF/HA/GRGD scaffolds incorporated with BMSC spheroids will be investigated as those performed in the patch. In the three-year project, we will develop dual bio-functional PCL-SF/HA/GRGD/ BMSC patch and further investigate the biomaterial properties such as grafting efficiency and surface analysis, and bioactivity analysis such as proliferation, differentiation properties of BMSC and formations of BMSC micro-mass on the patch examined by confocal microscopy and RT-PCR analysis. The same researches will be performed in scaffold/BMSC spheroid although the bioactivity of spheroids such as ability of cardiomyogenesis shall be first studied. For animal studies, the cardiac remodeling, apoptosis, angiogenesis, paracrine factors analysis in the MI zones will be performed as early investigations by this team. This study provides an important base for the research cardiac repairs of MI hearts of cardiac patch and scaffolds/BMSC to further perform pre-clinical studies.