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Objective: To evaluate the effect of transplanting hone marrow-derived mesenchymal stem cells (BM-MSCs) or adenovirus5-hepatocyte growth factor (Ad5-HGF) via non-infarct-related artery injection in swine myocardial infarction models. Methods BM-MSCs were obtained from swine bone marrow and expanded in vitro to a purity of ＞50%. A myocardial infarction (MI) was created by ligating the distal left anterior descending artery in swine. Either BM-MSCs (5×10^6/ml) or Ad5-HGF (4×l0^9 pfu) were transfused via the right coronary artery (non-infarcted artery) four weeks after ML Gate-controlled cardiac perfusion imaging was performed at the end of four and seven weeks after LAD ligation, to evaluate heart function and cardiac perfusion. Morphologic and histologic characteristics of the hearts were also studied. Results: (1) The gate-controlled cardiac perfusion imaging showed that the improvement in LVEF was greater in both treatment groups than in control group at the 4(superscript th) weeks. (2) In both treatment groups, capillary density was significantly higher than that of control group (P＜0.05). Conclusion: BM-MSCs or Ad5-HGF transplantation via non-infarcted artery administration can stimulate angiogenesis and improve heart function, but there was no difference in therapeutic efficacy between BM-MSCs and Ad5-HGF.

We investigated the impact of bone marrow-derived mesenchymal stem cells (BM-MSCs) alone or in combination with hepatocyte growth factor (HGF) transplantation via noninfarct-relative artery in a swine myocardial infarction (MI) model. Donor BM-MSCs were derived in vitro from swine auto-bone marrow cultures labeled by bromodeoxyuridine (BrdU) incorporation. Host MI swine model was created by ligating the distal left anterior descending artery. After 4 weeks, age-matched male MI swines were used for the transplantation. Male MI swines were transfused via noninfarct-relative artery with vehicle (control, n=6) or BrdU-labeled BM-MSCs (5 x 10(6)) alone (MSCs, n=6) or BrdU-labeled BM-MSCs (5 x 10(6)) combined with HGF (4 x 10(9) PFU) (MSCs+HGF, n=6). To evaluate the collateral artery growth (Rentrop) and cardiac perfusion in these animals, gate cardiac perfusion imaging and coronary angiography were performed before and 4 weeks after transplantation, respectively. To assess the contribution of donor-originated cells in stimulation of cardiomyocyte regeneration and angiogenesis, immunohistochemistry for BrdU and alpha-smooth muscle actin (alpha-SMA) and quantitative image analysis were performed at 4 weeks after transplantation. The results are as follows: (1) BrdU-positive cells were detected in host myocardium in both MSCs and MSCs+HGF groups, but not in the vehicle group. Most BrdU-positive cells expressed myosin heavy chain beta. (2) alpha-SMA(-)positive arteriole densities in the infarcted border area and infarcted area were increased significantly in both transplantation groups compared with the vehicle group. (3) Gate cardiac perfusion imaging demonstrated that the cardiac perfusion was significantly improved in transplantation groups compared with the vehicle group. (4) Ejection fraction and alpha-SMA-positive arteriole densities were increased significantly in both transplantation groups compared with the vehicle group. However, there was no difference in ejection fraction and alpha-SMA-positive arteriole densities between the MSCs group and the MSCs+HGF group. Growth of collateral arteries was not detected by coronary angiography in all three groups. In conclusion, the current study indicates that BM-MSCs transplantation via noninfarct-relative artery stimulates cardiomyocyte regeneration and angiogenesis and improves cardiac function, but does not stimulate collateral artery growth. BM-MSCs transplantation combined with HGF therapy is not superior to BM-MSCs alone transplantation. BM-MSCs transplantation via noninfarct-relative artery may be an alternative for those patients who cannot be transplanted via infarct-relative artery in clinical practice.

To study the effect of adenovirus5-mediated human hepatocyte growth factor (Ad5-HGF) transfer on post-infarct heart failure in a swine model. Twelve young Suzhong swine were randomly divided into 2 groups: the Ad5-HGF group (n=6) and the null-Ad5 group (n=6). Four weeks after left anterior descending coronary artery (LAD) ligation, Ad5-HGF was transferred into the myocardium via the right coronary artery. Coronary angiography and gated cardiac perfusion imaging were performed at the end of 4 and 7 weeks after LAD ligation, respectively, to evaluate collateral artery growth and cardiac perfusion. Then all animals were killed, the expression of HGF and α-smooth muscle actin (α-SMA) were evaluated by enzyme-linked immunosorbent assay and immunohistochemistry. Compared with the null-Ad5 group, higher expression of human HGF was observed in the myocardium in the Ad5-HGF group (109.3±7.8 vs 6.2±2.6, t=30.685, P

Despite advances in surgical and reperfusion therapy, there is no effective therapy currently exists to prevent the progressive decline in cardiac function following myocardial infarction. Hepatocyte growth factor has potent angiogenic and anti-apoptotic activities. The aim of this study was to investigate the therapeutic effect and dose-effect relationship on postinfarction heart failure with different doses of adenovirus-mediated human hepatocyte growth factor (Ad(5)-HGF) transference in swine models. Totally twenty swine were randomly divided into four groups: (a) control group (null- Ad(5), 1 ml); (b) low-dose group (1 x 10(9) Pfu/ml Ad(5)-HGF, 1 ml); (c) medium-dose group (5 x 10(9) Pfu/ml Ad(5)-HGF, 1 ml); (d) high-dose group (1 x 10(10) Pfu/ml Ad(5)-HGF, 1 ml). Four weeks after left anterior descending coronary artery (LAD) ligation, different doses of Ad(5)-HGF were transferred in three therapeutic groups via right coronary artery. Four and seven weeks after LAD ligation, gate cardiac perfusion imaging was performed to evaluate cardiac perfusion and left ventricular ejection fraction (LVEF). Seven weeks after surgery, the apoptotic index of cardiocyte was observed by TUNEL, the expression of Bcl-2, Bax, alpha-SMA and Factor VIII in the border zones were evaluated by immunohistochemistry, respectively. Four weeks after myocardial infarction, no significant difference was observed among four groups. Three weeks after Ad(5)-HGF transfer, the improvement of cardiac perfusion and LVEF was obviously observed, especially after 1 x 10(10) Pfu Ad(5)-HGF transfer. TUNEL assay showed that 5 x 10(9) Pfu and 1 x 10(10) Pfu Ad(5)-HGF treatment had a obvious reduction in the apoptotic index compared with the null-Ad(5) group, especially after 1 x 10(10) Pfu Ad(5)-HGF treatment. The expression of Bcl-2 protein was increased and the expression of Bax protein was inhibited in the 5 x 10(9) Pfu and 1 x 10(10) Pfu Ad(5)-HGF groups compared with the null-Ad(5) group. The vessel density of Factor VIII(+) and alpha-SMA(+) was increased in Ad(5)-HGF groups compared with the null-Ad(5) group. There were no significant differences in angiogenesis, reducing apoptosis and ameliorating heart function between the 1 x 10(9) Pfu Ad(5)-HGF group and the null-Ad(5) group. Although no statistical difference was observed between 1 x 10(10) Pfu and 5 x 10(9) Pfu Ad(5)-HGF groups, the cardiac protective effects of 1 x 10(10) Pfu Ad(5)-HGF treatment were greater than 5 x 10(9) Pfu Ad(5)-HGF treatment. Different doses of Ad5-HGF injected via noninfarct-related artery could induce angiogenesis, reduce apoptosis and ameliorate heart function, and the cardiac protective effects of 1 x 10(10) Pfu Ad5-HGF is of most significance.

SUMMARY 1 There is growing evidence of the beneficial effects of hepatocyte growth factor (HGF) in myocardial infarction, heart failure and occlusive peripheral arterial disease. The aim of the present study was to evaluate the effects of intracoronary administration of an adenovirus vector encoding the human HGF gene (Ad-HGF) on serum levels of cytokines and mobilization of CD34+ and CD117+ cells in patients with coronary heart disease. 2 Twenty-one patients with severe coronary artery disease were recruited to the study: 11 patients received both a stent and administration of Ad-HGF; the remaining 10 patients received a stent alone and served as the control group. Blood samples were obtained from the femoral vein before and then 6 and 24 h, 3 and 6 days and 2 weeks after treatment for the isolation of serum and peripheral blood mononuclear cells. Intracoronary administration of Ad-HGF in patients with coronary heart disease resulted in high levels of HGF gene expression, as well as its receptor c-met, compared with the control group, as demonstrated by real-time reverse transcription–polymerase chain reaction. In addition, serum levels of HGF, vascular endothelial growth factor, monocyte chemoattractant protein-1 and interleukin (IL)-10 were increased and serum levels of IL-8 were decreased in patients administered Ad-HGF compared with the control group. The percentage of CD34+ and CD117+ cells in the peripheral blood increased in patients administered Ad-HGF. 3 In conclusion, HGF gene therapy may play an important role in the regulation of cytokines and the induction of endothelial progenitor cell mobilization in patients with coronary heart disease.

We aim to study the amelioration effect of adenovirus5-mediated human hepatocyte growth factor gene transfer on postinfarction heart failure in swine model. Twelve Suzhong young swine were randomly divided into 2 groups of 6 pigs each: Ad(5)-HGF group and mock-vector Ad(5) group. Four weeks after ligation of the left anterior descending coronary artery, Ad(5)-HGF was intracoronarily transferred into the myocardium. Simultaneously, gate cardiac perfusion imaging was performed to evaluate the heart function. Three weeks later, gate cardiac perfusion imaging was performed again, then the hearts were removed and sectioned for immunohistochemical examination to illustrate the effects of Ad(5)-HGF on infarcted myocardium. The expression of HGF was examined by ELISA. The results were: (1) compared with the mock-vector Ad(5) group, high expression of human HGF was observed in the myocardium of Ad(5)-HGF group; (2) in the Ad(5)-HGF group, the number of CD117(+) cells co-expressing c-Met per mm(2) was significantly larger; (3) the improvement in LVEF was greater in the Ad(5)-HGF group than in the mock-vector Ad(5) group. We concluded that: (1) high expression of human HGF was observed in the myocardium through intracoronary gene transfection; (2) HGF can improve the mobilization of CD117(+)/c-Met(+) stem cells into ischemic myocardium. The amelioration effect of HGF on postinfarction heart failure could not be limited to stimulating angiogenesis, anti-apoptosis, anti-fibrosis, but was also involved in the recruitment of stem cells into myocardium.

Adult bone marrow stromal cells could differentiate into myogenic endothelial progenitor cells and has been investigated for the potential value in regeneration. Recently, it has been reported that bone marrow cells (BMCs) are able to repair the infracted myocardium by intracoronary transplantation via infarct-related artery in humans. Unfortunately, we cannot open the infarcted artery by traditional reperfusion therapies in some patients. We investigate the hypothesis that BMCs transplantation might get the same effect via noninfarct-relative artery. This alternative approach may have potential application in clinical practice.A swine myocardial infarction model was established by distal left anterior descending artery ligation. Bone marrow stromal cells isolated, culture-expanded and labeled with bromodeoxyuridine (BrdU) were used as donor cells. Four weeks after coronary artery ligation, either a graft of 5x10(6) donor cells (n=12) or culture medium (n=6) was infused into infarcted area via infarct-relative artery (left coronary artery, n=6) and noninfarct-relative artery (right coronary artery, n=6). Heart function was evaluated by gate cardiac perfusion imaging before the transplantation and 4 weeks after transplantation. The donor cell localization and differentiation were identified by immunohistochemical staining for BrdU and beta-myosin heavy chain (beta-MHC) and angiogenesis was assessed by immunohistochemical staining for alpha-smooth muscle actin (alpha-SMA) and Factor VIII.Gate cardiac perfusion imaging demonstrated that the cardiac function was significantly improved after the stromal cell transplantation via both infarct-relative and noninfarct-relative coronary arteries compared with control group (45.03+/-2.71 and 47.78+/-2.64 vs 30.36+/-2.76, P0.05). Four weeks after transplantation, BrdU and beta-MHC positive cells were detected within the infarct area. Vessel densities in infarct area and infarct border area were increased significantly in both transplantation groups compared to the control group (98.68+/-5.32 and 87.49+/-6.04 vs 48.46+/-4.88, P0.05).Transplantation of bone marrow stromal cell via both infarct-relative and noninfarct-relative coronary arteries improved heart function in the myocardial infarction animals by stimulating cardiomyocyte regeneration and angiogenesis.

To evaluate the impact of combined therapy with transplanting bone marrow-derived mesenchymal stem cells (BM-MSCs) via noninfarct-relative artery and hepatocyte growth factor (HGF) in a porcine myocardial infarction (MI) model.BM-MSCs were obtained from pig bone marrow, expanded in vitro with a purity of50%. MI was induced by ligating the distal left anterior descending artery in pigs. Eighteen animals received BM-MSCs cells (5 x 10(6)/ml, n = 6), BM-MSCs cells (5 x 10(6)/ml) plus HGF (4 x 10(9) pfu, n = 6) or equal volume culture medium (IMDM) via non-infarct-related artery at four weeks after MI. Gated myocardial perfusion imaging and coronary angiography were performed before and four weeks after transplantations. Histological examination was also performed 4 weeks after transplantation.LVEF measured by gated myocardial perfusion imaging was similar among groups before transplantation and significantly increased in BM-MSCs (45 +/- 3 vs. 34 +/- 2%, P0.05) or BM-MSCs + HGF (46 +/- 6 vs. 34 +/- 3%, P0.05) treated animals while remained unchanged in IMDM (30 +/- 3 vs. 32 +/- 2%) treated animals 4 weeks post transplantation. Similarly, capillary density was also significantly higher and myocardial perfusion defect scores significantly decreased in BM-MSCs or BM-MSCs + HGF treated hearts than that in IMDM treated hearts. However, all these changes were similar between BM-MSCs and BM-MSCs + HGF groups. Rentrop score was similar before and 4 weeks after transplantation among various groups.HGF in combination with BM-MSCs transplantation did not enhance the cardiac repair effects of BM-MSCs transplantation alone and BM-MSCs transplantation did not improve collateral circulation in this model.