| The application of vascular stents, as one of the interventional therapies that can cure the cardio-vascular disease, has been great developed since the vascular stents came out. And many researchers, doctors and other relevant experts have recently been focusing on developing cell seeding stents, which can be used as vehicles for prolonged and sufficient intramural drug delivery. It is firstly spraying a layer of medium on the surface of stent which is the carrier of cells, then embedding into body after seeding transgenic cells on the surface, which could cure the damaged endothelium in atherosclerotic therapies. There has been no cell seeding stent coming into domestic and overseas market at present, and it showed that there were still many disadvantages in researches. Just because the advantages of cell seeding stent in atherosclerotic therapies, great demanding in domestic market and monopolization by overseas corporation, it is important for our country to own home-developed and home-made cell seeding stent, which has lower price and higher quality.The achievement of a hospitable relationship between stent, coating matrix, endothelial cells and vessel wall is extremely challenging, so it is necessary that all the relevant knowledge should be applied comprehensively to solve the varied problems that would occurred in the process of developing cell seeding stent. The problems in developing cell seeding are focusing on long preparation periods, adhesion inhomogeneity between cells and stents, deficiencies of cells seeding on the stent, loose adhesion between cells and stent and so on. In order to develop cell seeding stent, the rotating culture device was invented, and the glutin was chosen as matrix on the surface of stent while ECV304 was the cell chosen, and rotating culture was used as the principle, and contrast the cells seeded on the stent with the cells which was static cultured by the NO expression and cell cycle to show the differences between them at last.Firstly, according to rotating culture principle and references, the automatic rotating culture device was designed and invented. The device contains six parts which were rotating culture tube, supporting tube, electromotor, reducer, connecting pole and turntable. The rotating tube was the space where the cells were adequately contacting with the stent; the supporting tube was to connect the stent; electromotor was used to provide the electro-drive force; reducer was used to reduce the rotating speed which was suitable for the cell culture; connecting pole was used to connecting the reducer and turntable; and the turntable was used to drive the rotating culture tube to rotate.Secondly, technological process of developing cell seeding stent by rotating culture was introduced in this text. Glutin was sprayed onto the surface of stent before it was placed in the rotating culture device. Considering the factors of influencing adhesion between cells and stent including rotating speed, rotating time, cell density and rotating times, calculating the best condition by counting the cells adhering to the stent; the stent surface was observed by optical microscope and scanning electron microscope(SEM) to confirm if the result of cells seeded on the stent was best.Finally, it was compared between rotating culture cells on the stent surface and static culture cells by NO expression and cell cycle after the cell seeding stent was prepared, which could judge the effect of the cell seeding stent.The experimental result revealed that a number of cells adhered firmly and equably to the surface of cell seeding stent prepared by self-design rotating culture device and optimized technological process; the optimize condition was calculated which was 0.4rpm, 6h,1×106 cells/ml, rotating 1 time, suitable for embedding into body. The expression of NO of rotating culture was more than static culture cells after 12h, while cell cycle was no change, revealing that the cell seeding stent was successfully prepared. |
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