Perfusion promotes endothelialized pore formation in high concentration fibrin gels otherwise unsuitable for tube development

Vascularization of tissue engineered implants is crucial for their survival and integration in the recipient’s body. Pre-vascularized, fibrin-based implants offer a solution since low concentration fibrin hydrogels (1 mg/mL) have been shown to promote tube formation of endothelial cells in co-culture with adipogenic stem cells. However, higher fibrinogen concentrations (> 20 mg/mL) enabling the fabrication of stable implants are necessary.

We here characterized fibrin gels of 1–30 mg/mL for their rheological properties and whether they support tube formation of endothelial cell–adipogenic stem cell co-cultures for up to 7 days. Moreover, 20 mg/mL gels containing preformed channels and endothelial cell–adipogenic stem cell co-culture were perfused continuously in a customized flow chamber with 3.9 dyn/cm² for 12 days and analyzed for capillary formation.

Rheology of fibrin gels showed increasing stability proportional to fibrinogen concentration with 20 mg/mL gels having a storage module of 465 Pa. Complex tube networks stable for 7 days were observed at 1–5 mg/mL gels whereas higher concentrations showed initial sprouting only. However, perfusion of 20 mg/mL fibrin gels resulted in endothelialized pore formation in several layers of the gel with endothelial cell–adipogenic stem cell co-culture.

Thus, perfusion supports the formation of capillary-like structures in fibrin gels that are too dense for spontaneous tube formation under static conditions. Future studies are necessary to further increase pore density and to investigate proper nutrition of tissue-specific target cells in the scaffold.