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经皮植入记忆合金自体膨胀支架体外研究.ppt

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1、2018/11/3,2008 CSIR Beijing Hongjian SHI,Implantation of Endothelial Progenitor Cells on a Self-expanding Metal Stent: an in vitro Study,Hongjian Shi,MD Gaojun Teng,Ph.D,MD Aihong Cao,MD Deparment of Radiology, Zhongda Hospital, Southeast Universty,2018/11/3,2008 CSIR Beijing Hongjian SHI,Content,Ba

2、ckground Materials and methods Results Discussion,2018/11/3,2008 CSIR Beijing Hongjian SHI,Background,Restenosis:pseudointimal hyperplasia ( PTA, stent) High restenosis in TIPS(30%-50%, 1y) Bile leakage unavoidable during TIPS Bile inhibit both SMC and EC Covered Viattor stent1. Coldwell D, Radiolog

3、y 19952. Zhuang Z,AJR 20023. LaBerge J,JVIR 19934. Nishimine K,Radiology 19955. Teng G,Radiology 1998;20056. Cejna M,Radiology 20017. Hausegger K,JVIR 2004,2018/11/3,2008 CSIR Beijing Hongjian SHI,Background,EPC(endothelial progenitor cell) can differentiate into EC(endothelial cell) and incorporate

4、 into sites of active angiogenesis. Asahara, Science 1997,2018/11/3,2008 CSIR Beijing Hongjian SHI,Shantsila E, et al.JACC 2007,2018/11/3,2008 CSIR Beijing Hongjian SHI,Zammarettia P, Int J Biochem Cell Biol 2005; 37: 493-503.,2018/11/3,2008 CSIR Beijing Hongjian SHI,EPC Isolation,Culture,Identifica

5、tion,Healthy domestic farm swine(25kg) Density gradient centrifugation Culture medium- EGM-2 Cell markers CD34, VEGFR-2, Vwf Specific function: uptaken DiI-AC-LDL,and binding FITC-UEA-1Ben-Shoshan, PharmacologyTherapeutics 2007,2018/11/3,2008 CSIR Beijing Hongjian SHI,Representative relative distrib

6、ution of some of the antigens expressed in CECs, as depicted by flow cytometry.,Bertolini F et al. Drug Discovery Today 2007;12:806-812,2018/11/3,2008 CSIR Beijing Hongjian SHI,Photographs of EPC,2018/11/3,2008 CSIR Beijing Hongjian SHI,EPC uptaken DiI-Ac-LDL,EPC binding FITC-UEA-1,2018/11/3,2008 CS

7、IR Beijing Hongjian SHI,Immunocytochemistry,EPC cell markers CD34, KDR,vWF,2018/11/3,2008 CSIR Beijing Hongjian SHI,Fibrinogen Preparation,15 ml peripheral blood from the same animal Cryoprecipitated techniqueCentrifugation at 1250 rpm at 4 for 20 min the plasma frozen to 70, stored at -20 4 refrige

8、rator centrifugation at 2500 rpms at 2 for 40 min 1.5ml concentrated fibriongenDePalma L, Transfusion 1993,2018/11/3,2008 CSIR Beijing Hongjian SHI,Procedure of EPC seeding on stent,10 self-expanding woven metal stents(Microtech, Nanjing): 5 in case group and 5 in control group. Fibrin gel(fibrinoge

9、n 5mg/ml+thrombin 0.1U/ml) Incubate for 24 h (rotate every 8 h) in EGM-2 with EPC concentration 1106 /ml Cell count Rinsed EPC re-identified1 Kipshidze N, JACC 20002 Shirota T, Biomaterials 2003,2018/11/3,2008 CSIR Beijing Hongjian SHI,2018/11/3,2008 CSIR Beijing Hongjian SHI,Cell count,Rinsed cell

10、from EPC seeded stent:2105,2018/11/3,2008 CSIR Beijing Hongjian SHI,Discussion,It is feasible to devise an EPC-seeded stent in vitro Applicabe in TIPS as well as vascular system EPC-seeded stent with photocured-gel CD34 antibody capture EPC Stent1 Shirota T, Biomaterials 20032 Aoki J, JACC 2005,2018

11、/11/3,2008 CSIR Beijing Hongjian SHI,Acknowlegement,This study was supported by grant 30670604 of the national natural science foundation of China Stents were sponsored by( Microtech Co.,Nanjing) Thanks for my collegues of Jiangsu Province key Lab of Molecular Imaging and Functional Imaging,2018/11/3,2008 CSIR Beijing Hongjian SHI,Thank you for your attention!,

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