In vitro construction of artificial blood vessels using spider silk as a supporting matrix.

ABSTRACT

Recent research has vigorously pursued the development of tissue-engineered vascular grafts (TEVs). One of the striking points of the early phase is the instability of the artificial blood vessels and lack of vascular resistance, which is supposed to be a consequence of unstable scaffolds. Therefore, alternative biological approaches are necessary to improve the physical properties of the artificial vessel walls. We developed blood vessel-like constructs based on native spider silk as a scaffold. C2C12 and ST1.6R cells were seeded on the surface of scaffolds and cultivated under pulsatile flow (max. ~135  mmHg and min. ~90  mmHg) in a bioreactor. Constructed grafts were compared to human blood vessels and cell seeded scaffolds cultured in the absence of bioreactor. Mechanical properties, morphological structures and expression of marker genes were assessed by strength and strain experiments, SEM, histological staining, immunohistology, Western blotting and quantitative real-time PCR. The results indicate that the constructed vessel resembles native blood vessels in morphological structure as well as in function and expression of biomarkers. Spider silk scaffolds seem to provide an optimal and stable basis for vessel constructs.