Experimental Study Of The Vessel Density And Collagen Content Of Autologous Mesenchymal Stem Cell Transplantation In Myocardial After Acute Myocardial Infarction

Objective:To establish thoracotomy coronary artery ligation dogs acute myocardial infarction(AMI) animal model,the injection means of myocardial infarction in the marginal dogs implanted autologous mesenchymal stem cells(BMMSCs),observed before and after transplantation in vascular endothelial growth factor(VEGF) concentration of changes,changes in capillary density and myocardial collagen fiber content changes of the AMI BMMSCs ventricular remodeling after the impact.Methods:The experiment was 2006-6/2007-12 in the First Affiliated Hospital of Anhui Medical University,the animal operating room,the center of Anhui Medical University animal laboratory and the completion.①Healthy dogs 40,13～18(15.5±3.2) kg,male and female informal.Were randomly divided into two groups:the transplantation group 20 and the control group 20.Transplant group and the control group is divided into four subgroups,respectively,1 w group,2 w group,3 w group and 4 w group.②Cell transplantation before 2～3 wk,healthy dogs after general anesthesia after the iliac crest bone marrow puncture access to 10 ml,with 1.073 g/ mLPereoll centrifugal liquid separation,to LG-DMEM medium plus 10% FBS culture.③Inverted microscope daily observation of cell growth,regular exchange fluid,spread to two or three generations after the transplant before 1 d cell transplantation will be tagged with BrdU.④Ligation dogs left anterior descending coronary artery(LAD) after the establishment of AMI Model 1～2 hr,BrdU transplantation group will be marked BMMSCs(5×10~6 cells) 2 ml suspension infarct area edge into four different parts of the control group to adopt the same approach into the equivalent DMEM.⑤Respectively after transplantation 1 d and 1,2,3, 4wk collected blood,after detaining centrifugal serum,-70℃preservation reserve.⑥Transplant after 4 wk to ELISA(ELISA) in serum concentrations of VEGF; animals were killed to obtain samples myocardial infarction zone,slice after anti-BrdU BMMSCs immune staining observed in memory of myocardial live,at the same time lineⅧfactor polyclonal antibody immunohistochemical staining observed in the cardiac and vascular density MASSION three-color stain of collagen fiber content.Results:①40 dogs,both succeeded in establishing a model of AMI,survived to the end of experiments.②BMMSCs training of dogs in good condition,after the passage BrdU tagging,tagging rate of 80 percent.③Transplant group and the control group were increased serum VEGF,2～3 wk up to the peak,but higher rate of transplantation is greater than the control group(p＜0.05).Although the 3 wk after transplantation VEGF and control groups showed a downward trend,but the transplantation group VEGF show marked higher(p＜0.01).④Myocardial infarction biopsy found that compared with the control group,cardiac transplantation within the group to see more positive staining BrdU immune cells;units of blood vessels significantly increased(p＜0.01),collagen fibers were significantly reduced(p＜0.05),peripheral vascular collagen deposition reduction.Conclusion:The dogs AMI,the myocardial injection means the implantation of autologous BMMSCs in myocardial infarction,colonization,survival,and to promote angiogenesis,the formation of collagen reduced,thus benefiting the delay ventricular remodeling after AMI.