Abstract
Due to its low immunogenic potential and the possibility to fine-tune their properties, materials made of recombinant engineered spider silks are promising candidates for tissue engineering applications. However, vascularization of silk-based scaffolds is one critical step for the generation of bioartificial tissues and consequently for clinical application. To circumvent insufficient vascularization, the surgically induced angiogenesis by means of arteriovenous loops (AVL) represents a highly effective methodology. Here, previously established hydrogels consisting of nano-fibrillary recombinant eADF4(C16) were transferred into Teflon isolation chambers and vascularized in the rat AVL model over 4 weeks. To improve vascularization, also RGD-tagged eADF4(C16) hydrogels were implanted in the AVL model over 2 and 4 weeks. Thereafter, the specimen were explanted and analyzed using histology and microcomputed tomography. We were able to confirm biocompatibility and tissue formation over time. Functionalizing eADF4(C16) with RGD-motifs improved hydrogel stability and enhanced vascularization even outperforming other hydrogels, such as fibrin. This study demonstrates that the scaffold ultrastructure as well as biofunctionalization with RGD-motifs are powerful tools to optimize silk-based biomaterials for tissue engineering applications.
Highlights
Large volume tissue defects, as a result of trauma or radical tumor resection, often require autologous tissue transfer for defect reconstruction
Modern reconstructive surgery underwent a continuous development with regard to less invasive surgical methods, autologous tissue transplantation can be associated with a considerable donor site morbidity
The recombinant spider silk protein eADF4(C16)-RGD was synthesized as a soluble protein in Escherichia coli BL21 gold (DE3) using a time-dependent fed-batch fermentation strategy as described previously (Huemmerich et al 2004, Wohlrab et al 2012)
Summary
As a result of trauma or radical tumor resection, often require autologous tissue transfer for defect reconstruction. Modern reconstructive surgery underwent a continuous development with regard to less invasive surgical methods, autologous tissue transplantation can be associated with a considerable donor site morbidity. The latter one and/or patient specific characteristics (such as previous operations, radiotherapy) can limit tissue availability. First described by O’Shaughnessy and later by Shen et al over four decades ago, the principle of flap prefabrication is a promising strategy to generate large vascularized tissues for defect reconstruction mainly in head/neck applications or burn surgery (O’shaughnessy 1936, Guo and Pribaz 2009). Besides the distal ligation of an artery with its venae comitantes creating an arteriovenous bundle, arteriovenous loops (AVL) are used for flap prefabrication
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