Treatment Of Myocardial Infarction After Reperfusion In Rats With Bone Marrow Mesenchymal Stem Cells Induced By BMP-2

ObjectiveObservation of that whether the bone can form protein2(BMP-2) couldinduce bone marrow stem cells (BMSCs) which can differentiate into classbetween myocardial cells; Observation of BMSCs transplantation in rats withacute ischemic infarction reperfusion of myocardial function and the influence ofcardiac function of rat; Observed by the BMP-2preliminary induction andwithout inducing BMSCs into the therapeutic effect of acute infarctionmyocardial tissue, prompt experimental basis for the stem cell clinicalapplication.MethodsThe whole bone marrow adherent culture method separation of SD ratBMSCs in vitro amplification; Immune cell fluorescence method was applied tothe fourth generation of BMSCs identified; Take the fourth generation of BMSCsby BMP-2preliminary induction training after48hours in liquid, withconventional culture continues to develop. After4weeks of connection byimmunocytochemistry method43(CX-43) and myocardial specificity proteinexpression of troponin T (cTNT) analysis to identify the BMP-2BMSCsdifferentiation for the class in the process of the induced myocardial ability;BMSCs DAPI tag and fluorescent microscope. Under anesthesia SD ratmyocardial infarction reperfusion mode, building successful rats were randomlydivided into three groups. The control group into culture medium; Stem cellgroup implanted by DAPI labelled BMSCs, induced group implanted with DAPIlabeled after induced BMSCs;4hours after transplantation,4days,4weeksmyocardial based on slice observation DAPI labelled BMSCs after implantsurvival and distribution;4weeks using ultrasound examination in rats after transplantation LVEF, FS, and through the changes of myocardial tissueMasson and HE slice staining to observe the myocardial tissue pathologicalchanges.ResultsThe morphological observation and identification of BMSCs: primary cellassumes the circular or elliptic, more clear boundaries, has the refraction,suspended in a culture;3days most of the adherent growth, is in the shape ofpolygon and circle, spindle cells around the visible length, different degree ofswelled. A week or so, the cell division reproduce rapidly, the volume increasesgradually, the number increased significantly, irregular arrangement, form classlong spindle, a colony of samples gathered to grow. The4th generation cells,size, shape before reaching consensus, arranging a neat, formed a long spindle,like fibroblasts. Induced cells grow well after induction, morphologicalcharacteristics change gradually over time, the induction of4weeks, time didnot induce the pre induced cells increased significantly wider, size, shape ismore short rod, arrange contorts converged, compact connection, structurevisible muscle filaments, began to show the characteristics of the muscle cells.Identification of BMSCs,4th generation BMSCs surface antigen CD90, theexpression ofCD90and CD105was positive, positive rate were96.56%adc95.46%respectively, hematopoietic stem cell surface antigen CD34, CD45were negative expression, for the characteristics of BMSCs. After pre inducedBMSCs cTnT and the expression of CX43、BMSCs after BMP-2pre induced the4w, fluorescence microscope CX43, with vision cTnT in purple fluorescentgreen fluorescence, positive expression marks, without inducing BMSCs fromthe expression of CX43, cTnT negative; BMSCs induced after4weeks,differentiate into myocardial cells. Positive cardiac muscle specific proteinexpression.BMSCs tag and track after implantation: BMSCs after DAPI markers, aizennucleus and cytoplasm,dye,mark rate100%. By DAPI labeled BMSCs inpostoperative4hours,4days,4weeks, on the fluorescence microscopymyocardial tissue biopsies to induce group and stem cells have the DAPIlabeled BMSCs survived in the infarction myocardial tissue, and the survival of pre induced group stem cell group.Stem cell transplantation therapy of myocardial infarction: stem cells afterimplantation can live in myocardial reperfusion infarction environment; Stem celltransplantation for infarction myocardial reperfusion therapy effect; Stem cellsand induced beforehand group compared with control subjects left heartejection fraction (LVEF) and left ventricular shortening fraction (FS) wereincreased (P<0.01), and induced group is more apparent. Conclusionmyocardial tissue pathological slices: HE tissue staining control inflammatorycell infiltration is the most serious infarction area, stem cell group ofinflammatory cell infiltration is lighter. Inflammatory cells infiltrating the lightestinduced group. Control group infarction area with severe fibrosis, infarction is alarge patch of distribution continuity. BMSCs group and pre induced fibrosislighter infarction area. The lightest pre induced group degree of fibrosis.Control group infarction area with severe fibrosis, infarction is a large patchof distribution continuity. BMSCs group and pre induced fibrosis lighter infarctionarea. The lightest pre induced group degree of fibrosis. Myocardial tissuesection by Masson staining of BMSCs transplantation appear on collagenfibrous tissue. System observation4days to4weeks slices that collagen fiberhas no obvious change, and transplanted cells survived in the collagen fibers.ConclusionsBMSCs in vitro induced to the BMP-2kind of cardiac muscle celldifferentiation, BMSCs can be used to treat acute myocardial infarction (AMI)and reperfusion damage have a certain tolerance treatment effect, the stemcells induced by BMP-2pre treatment effect induced by less ideal.