Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/3474
標題: Dynamic endothelial cell seeding, culture and physical stimulation for poly(carbonate)urethane small-diameter vascular grafts
內皮細胞動態植覆、培養與物理刺激於碳酸酯型聚胺酯人工血管之研究
作者: Tsai, Yi-Jing
蔡易瑾
關鍵字: PCU;聚碳酸酯型聚胺酯;ultrasound;laser;dynamic seeding;dynamic culture;vWF;超音波;雷射;動態植覆;動態培養;vWF
出版社: 化學工程學系
摘要: 
The biomedical grade poly(carbonate)urethane synthesized in our lab was used to fabricate the small-diameter vascular grafts by salt-impregnating. The grafts had open-cell structure with a compliance similar to that of native vessels, conformed by SEM and mechanical tests.
To improve the seeding efficiency, an RGD coating protein, was coated on the internal surface of grafts before cell seeding, and the endothelial cells were pretreated stimulated by ultrasound or laser stimulation. It was found that efficiency was not enhanced by ultrasound or laser. However the endothelial cell retention after 3-hr flushing experiment was CBD-RGD. Ultrasound stimulation could increase the ability of endothelial cells to adhere on the internal surface of grafts, and make them more resistant to the shear force of the fluid. It was also shown by SEM and GAG analysis that endothelial cells cultured in the dynamic condition secreted more extracellular matrix to confront with the force of fluid. For endothelial cells cultured in the static condition, the cell activity and functionality might decrease with the increasing times of subculture.
It was found that the optimized culture and processes conditions of vascular grafts for implantation in animal body were: (1)before seeding the endothelial cells were expanded and stimulated by ultrasound for a week; (2)the cells seeded on the microporous PCU+gelatin+RGD grafts by dynamic rotational 0.16rpm seeding; (3)seeded grafts were cultured statically for 72hours and dynamically (perfusion type) for a week.

本研究嘗試引用自行合成的聚碳酸酯型聚胺酯(PCU)作為人工血管的材料,以混鹽浸鑄法的方式,製作出具微孔相連結構且符合天然血管順應性的人工小血管。由機械性質測試與SEM的照片證實。
改進動態植覆的方式,探討物理性超音波與雷射的刺激和CBD-RGD於細胞植覆的應用。超音波與雷射刺激並無明顯增加細胞的植覆率,而CBD-RGD改質對內皮細胞的貼附在短時間內有較佳的效果。內皮細胞在適當的超音波刺激下,有增進細胞更牢固貼附於材料表面的能力之趨勢,較不易因流體的作用力,從人工血管材料表面脫落。GAG分析與SEM的照片顯示,內皮細胞在動態環境下生長,為了抵抗流體的作用力,細胞會分泌較多的基質。由內皮細胞vWF鑑定中發現,內皮細胞在靜態環境下培養,其活性與功能可能會隨著代速的增加而逐漸的退化。
未來,對於最適合植入動物體內的人工小血管,其內皮細胞需經一週的培養與超音波的刺激,以動態(旋轉式,0.16rpm)的植覆方式植覆於porous PCU+gelatin+RGD的人工血管上,再給予靜態培養72小時,最後再置於動態(灌流式,9cm/sec)環境中培養一週。
URI: http://hdl.handle.net/11455/3474
Appears in Collections:化學工程學系所

Show full item record
 

Google ScholarTM

Check


Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.