EXPERIMENTAL USE OF A MODIFIED FIBRIN GLUE TO INDUCE SITE-DIRECTED ANGIOGENESIS FROM THE AORTA TO THE HEART

From 10 cultures of manipulated Escherichia coli bacteria expressing the class I heparin-binding growth factor polypeptide alpha-endothelial cell growth factor, 11.2 +/- 0.7 mg alpha-endothelial cell growth factor was eluted by heparin-sepharose affinity chromatography. Analysis of molecular weight (17,000 kD) was done by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and purification of the growth factor was done by high-performance liquid chromatography. The harvested alpha-endothelial cell growth factor was proved by protein blotting. To assess the growth-promoting activity, we did an endothelial cell growth assay by comparing adult human endothelial cell control cultures, without adding growth factor to the culture medium, with adult human endothelial eel cultures with 0.02 to 20.0 ng/ml alpha-endothelial cell growth factor and 1.0 ng/ml heparin and with adult human endothelial cell cultures with alpha-endothelial cell growth factor but without heparin. Tritiated thymidine counts proved the significant growth-promoting activity of alpha-endothelial cell growth factor. In 10 experimental animals modified fibrin glue containing 1 mu g alpha-endothelial cell growth factor was implanted between the aorta and the myocardium of the left ventricle and results were compared with those in five control animals that received normal fibrin glue without growth factor. After 9 weeks of implantation, angiography and histologic investigation showed newly grown vascular structures between the aorta and the myocardium in all experimental animals, but none in the control animals. Our study proved the feasibility of initiating site-directed formation of new blood vessel structures to the heart by a modified fibrin glue implant containing angiogenic growth factor alpha-endothelial cell growth factor.