Mária Giretová ^1* Ľubomír Medvecký ¹ Zuzana Demčišáková ² Lenka Luptáková ² Eva Petrovová ² Radoslava Štulajterová ¹

• ¹ Institute of Materials Research of Slovak Academy of Sciences, Košice, Slovakia
• ² University of Veterinary Medicine and Pharmacy in Košice, Košice, Slovakia

The aim of paper was to evaluate effect of gel adding to biopolymeric scaffolds on pro-angiogenic and basic material characteristics of the final composite for using in regenerative medicine. The studied scaffold consisted of natural biopolymers: polyhydroxybutyrate (PHB), chitosan, agarose and gelatine. The final scaffold was characterized by a high macroporosity (90 %) and wide pore size distribution. As is known, the pore size is a critical factor for cell ingrowth in grafts after implantation in body as well as for angiogenic development and creation of new vessels. After 9 days of cultivation in the culture medium, the scaffold retained its physicochemical properties without tendency to disintegration The addition of polymeric gels to scaffold improved the mechanical stability of composite. The in vitro cytotoxicity testing showed good adherence of the seeded L929 fibroblasts on scaffold and strong ingrowth of cells into the macropores. No sign of cytotoxicity was identified by both the MTS assay and live/dead cell staining. The quail chorioallantoic membrane (CAM) assay -as an alternative to in vivo assays, revealed suitable pro-angiogenic properties of scaffold for the formation and ingrowth of new blood vessels. Moreover, the upregulation of gene expression responsible for the activation of angiogenic cascade clearly demonstrated positive effect of prepared composites on angiogenesis as an essential part of the new tissue formation and the regeneration process itself.