Implantation of BM cells transfected with phVEGF₁₆₅ enhances functional improvement of the infarcted heart

Abstract

Background: Experimental data have suggested that BM-cell implantation can improve infarcted cardiac function. However, the number of implanted cells that survive is still unknown. The present study was performed to investigate whether implantation of autologous BM mononuclear cells (BM-MNCs) transfected with phVEGF₁₆₅ can increase the number of surviving implanted cells, and enhance functional improvement of infarcted hearts in rabbits.

Methods: Acute myocardial infarction (AMI) in rabbits was replicated by ligating the left anterior descending coronary artery, and animals were randomly divided into the following three groups: I AMI control group (n=7); II BM-MNCs transfected with phVEGF₁₆₅ implantation group (n=7); III BM-MNCs implantation group (n=7). In addition, sham-operated (n=5) rabbits were randomly selected to serve as a non-infarction control group (VI). Animals for cell implantation received intramyocardial injections of autologous BM-MNCs 14 days after AMI. Echocardiography and hemodynamic studies were performed to evaluate cardiac structure and function 28 days after implantation. The implanted sites were examined using immunofluorescence to identify the phenotypes and number of the labelled cells. Reverse transcriptase (RT)-PCR and Western blot analysis were performed to detect the expression of hVEGF₁₆₅ gene and VEGF₁₆₅ protein respectively.

Results: Failed cardiac function produced by AMI was significantly improved in Groups II and III 28 days after cell implantation. BM-MNCs transfected with phVEGF165 implantation conferred a further improvement of cardiac function, with significant changes of all assessed parameters when compared with BM-MNCs implantation alone (all P<0.05). The implanted cells demonstrated myogenic differentiation with the expression of Troponin T and organized contractile proteins. The positive staining for Factor VIII-related Ag indicated the induction of angiogenesis in the infarct area. The percentage of Brdu-positive myocyte, endothelial cells was 75±6%, 34.1±4.6% in Group II, significantly higher than that in Group III (51±7%, 11.3±2.5% respectively, P<0.05). RT-PCR analysis demonstrated exclusive expression of hVEGF165 gene in Group II, and Western blot showed the expression of VEGF165 protein was significantly higher in Group II than in Group III (P<0.05).

Conclusions: Implantation of BM-MNCs transfected with phVEGF₁₆₅ can increase the number of implanted cells surviving, and enhance the impaired cardiac function after AMI.