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1、Implantation of Endothelial Progenitor Cells on a Self-expanding Metal Stent: an in vitro StudyHongjian Shi,MDGaojun Teng,Ph.D,MDAihong Cao,MDDeparment of Radiology, Zhongda Hospital, Southeast Universty9/15/20242008 CSIR Beijing Hongjian SHIContentoBackgroundoMaterials and methodsoResultsoDiscussio
2、n9/15/20242008 CSIR Beijing Hongjian SHIBackgroundoRestenosis:pseudointimal hyperplasia ( PTA, stent)oHigh restenosis in TIPS(30%-50%, 1y)oBile leakage unavoidable during TIPSoBile inhibit both SMC and ECoCovered Viattor stent 1. Coldwell D, Radiology 1995 2. Zhuang Z,AJR 2002 3. LaBerge J,JVIR 1993
3、 4. Nishimine K,Radiology 1995 5. Teng G,Radiology 1998;2005 6. Cejna M,Radiology 2001 7. Hausegger K,JVIR 20049/15/20242008 CSIR Beijing Hongjian SHIBackground oEPC(endothelial progenitor cell) can differentiate into EC(endothelial cell) and incorporate into sites of active angiogenesis. Asahara, S
4、cience 1997 Asahara, Science 19979/15/20242008 CSIR Beijing Hongjian SHIShantsila E, et al.JACC 20079/15/20242008 CSIR Beijing Hongjian SHIZammarettiaZammarettia P, P, IntInt J J BiochemBiochem Cell Cell BiolBiol 2005;2005; 37: 493-503. 37: 493-503.9/15/20242008 CSIR Beijing Hongjian SHIEPC Isolatio
5、n,Culture,IdentificationooHealthy domestic farm swine(25kg)ooDensity gradient centrifugationooCulture medium- EGM-2ooCell markers CD34, VEGFR-2, VwfooSpecific function: uptaken DiI-AC-LDL,and binding FITC-UEA-1 Ben-Shoshan, Pharmacology Ben-Shoshan, PharmacologyTherapeutics 2007Therapeutics 2007 9/1
6、5/20242008 CSIR Beijing Hongjian SHIRepresentative relative distribution of some of the antigens expressed inCECs, as depicted by flow cytometry. Bertolini F et al. Drug Discovery Today 2007;12:806-8129/15/20242008 CSIR Beijing Hongjian SHIPhotographs of EPC9/15/20242008 CSIR Beijing Hongjian SHIEPC
7、 uptaken DiI-Ac-LDLEPC binding FITC-UEA-19/15/20242008 CSIR Beijing Hongjian SHIImmunocytochemistry EPC cell markers CD34, KDR,vWF9/15/20242008 CSIR Beijing Hongjian SHIFibrinogen Preparationo15 ml peripheral blood from the same animaloCryoprecipitated technique Centrifugation at 1250 rpm at 4 for 2
8、0 min the plasma frozen to 70, stored at -20 4 refrigerator centrifugation at 2500 rpms at 2 for 40 min o1.5ml concentrated fibriongen DePalma L, Transfusion 19939/15/20242008 CSIR Beijing Hongjian SHIProcedure of EPC seeding on stento10 self-expanding woven metal stents(Microtech, Nanjing): 5 in ca
9、se group and 5 in control group.oFibrin gel(fibrinogen 5mg/ml+thrombin 0.1U/ml)oIncubate for 24 h (rotate every 8 h) in EGM-2 with EPC concentration 1106 /mloCell count oRinsed EPC re-identified 1 Kipshidze N, JACC 2000 2 Shirota T, Biomaterials 20039/15/20242008 CSIR Beijing Hongjian SHI9/15/202420
10、08 CSIR Beijing Hongjian SHICell countoRinsed cell from EPC seeded stent: 21059/15/20242008 CSIR Beijing Hongjian SHIDiscussionoIt is feasible to devise an EPC-seeded stent in vitrooApplicabe in TIPS as well as vascular systemoEPC-seeded stent with photocured-geloCD34 antibody capture EPC Stent 1 Sh
11、irota T, Biomaterials 2003 2 Aoki J, JACC 20059/15/20242008 CSIR Beijing Hongjian SHIAcknowlegementoThis study was supported by grant 30670604 of the national natural science foundation of ChinaoStents were sponsored by( Microtech Co.,Nanjing)oThanks for my collegues of Jiangsu Province key Lab of Molecular Imaging and Functional Imaging9/15/20242008 CSIR Beijing Hongjian SHIoThank you for your attention!9/15/20242008 CSIR Beijing Hongjian SHI