ABSTRACT: Fibrin nanobeads were synthesized through a simple water-in-oil emulsification diffusion technique. Human vascular endothelial growth factor (VEGF) was added to the precursor fibrinogen solution before polymerisation and allowed to conjugate within the nanobeads during the process of emulsification. The nanobeads were recovered from the emulsion and freeze-dried to produce stable nano-formulation for prolonged in vitro studies. The in vitro degradation rate of nanobeads and subsequent release of VEGF was quantified through ELISA technique. The functionality of formulated nanobeads was assessed by endothelial proliferation and tube formation experiments. The nanobeads were then embedded among Agarose based 3-D scaffolds. The fibrin nanobeads as well as the scaffolds were visualized through SEM and AFM, analyzed for their physico-chemical properties through XRD, TGA, DLS, Zeta potential etc. and characterized by means of UV and IR spectroscopy. Human umbilical vein endothelial cells (HUVEC) were seeded on to the scaffolds and the in vitro vascularization attained through the controlled release of VEGF was visualized through fluorescent microscopy and confocal laser microscopy suggesting the potential of VEGF conjugated fibrin nanobeads as an ideal candidate for in vitro vascularization among 3D Tissue Engineered constructs.
4TH BANGALORE NANO; 01/2011