release kinetics of vegf165 from a collagen matrix and structural matrix changes in a circulation model从胶原蛋白释放动力学vegf165矩阵和结构矩阵循环模型的变化.pdfVIP

Kleinheinz et al. Head Face Medicine 2010, 6:17 /content/6/1/17 HEAD FACE MEDICINE R E S E A R C H Open Access Research Release kinetics of VEGF165 from a collagen matrix and structural matrix changes in a circulation model 1 1 1 2 1 Johannes Kleinheinz* , Susanne Jung , Kai Wermker , Carsten Fischer and Ulrich Joos Abstract Background: Current approaches in bone regeneration combine osteoconductive scaffolds with bioactive cytokines like BMP or VEGF. The idea of our in-vitro trial was to apply VEGF165 in gradient concentrations to an equine collagen carrier and to study pharmacological and morphological characteristics of the complex in a circulation model. Methods: Release kinetics of VEGF165 complexed in different quantities in a collagen matrix were determined in a circulation model by quantifying protein concentration with ELISA over a period of 5 days. The structural changes of the collagen matrix were assessed with light microscopy, native scanning electron microscopy (SEM) as well as with immuno-gold-labelling technique in scanning and transmission electron microscopy (TEM). Results: We established a biological half-life for VEGF165 of 90 minutes. In a half-logarithmic presentation the VEGF165 release showed a linear declining gradient; the release kinetics were not depending on VEGF165 concentrations. After 12 hours VEGF release reached a plateau, after 48 hours VEGF165 was no longer detectable in the complexes charged with lower doses, but still measurable in the 80 μg sample. At the beginning of the study a smear layer was visible on the surface of the complex. After the wash out of the prote