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GENE TRANSFER INTO PORCINE MYOCARDIUM VIA PERICARDIAL CAVITY BY HOMEMADE EASY PUNCTURE DEVICEA
Authors:Huai-min Guan  Peng Liu  Jin-hong Xie  Feng-ling Wang  Lin-sheng Cao  Qi-jun Qian
Abstract:Objective To explore the feasibility and safety of gene transfer into porcine myocardium via the pericardial cavity by a homemade easy device.Methods Replication-deficient recombinant adenoviral vector carrying LacZ report gene (Ad-LacZ) was constructed by the calcium phosphate precipitation method. Twelve healthy Chinese mini-swine were randomly divided into experimental group (n = 6) and control group (n =6). Acute myocardial infarction (AMI) model was established by balloon occlusion of the distal part of D[ branch of left anterior descending (LAD) artery, at the same time the intra-pericardial cavity injections were performed through the small incision of the abdominal wall below the xyphoid appendix using a homemade device. Then gene transfer was performed using a central venous catheter. The pericardium was pre-treated with injection of a mixture of collagenase (1 200 U) and hyaluronidase (3 000 U) in both groups. Then 2. 0 × 109 plaque formation unit (PFU) Ad-LacZ was injected into the pericardial cavity in experimental group, while 1 mL of normal saline was injected in the control group. The p-galactosidase activity detection and X-gal staining of the ischemic myocardium were performed on the 3rd, 7th, and 28th day after injection.Results The LAD artery was occluded completely and infarction and ischemia were detected by histological assessment In experimental group, the X-gal staining positive cells and (3-galactosidase activity quantification were detectable on the 3rd day after injection, increased markedly on the 7th day, and then declined on the 28th day. The transfer efficiencies indicated by the positive myocardial cells were 16. 7% , 45. 6% , 22. 8% on the 3rd, 7th, 28th day, respectively. In control group, no positive cells and (3-galactosidase activity were observed.Conclusion Adenovirus can be transferred into ischemic myocardium and express target gene in the AMI model for four weeks with the homemade easy device via pericardial cavity pretreated by collagenase and hyaluronidase.
Keywords:adenovirus  acute myocardial infarction model  gene transfer  pericardial cavity  PUNCTURE  EASY  PERICARDIAL CAVITY  MYOCARDIUM  PORCINE  TRANSFER  Adenovirus  transferred  express  target gene  four  myocardial cells  increased  quantification  positive  Results  infarction  ischemia  assessment  activity
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