The Next Step in In-Vivo Liver Engineering: Generation of an In vivo Liver Lobe Scaffold Which is Structurally Intact and Physiologically Reperfusable

Abstract

In-vivo liver decellularization has become a promising strategy to study in-vivo liver engineering. However, long term survival after in vivo liver decellularization has not yet been achieved. This study aims to establish a survival model of in-vivo perfusion-decellularization of a selective liver lobe.We compared three decellularization protocols (1%Triton X100 followed by1%SDS, 1% SDS versus 1%Triton X100, n=6/group). Using the optimal one as judged by macroscopy, histology and DNA-content, we characterized the structural integrity and matrix-proteins using Histology, scanning electron microscopy (SEM), CT scanning and Immunohistochemistry. We prevented contamination of the abdominal cavity with the corrosive detergents by using polyvinylidene chloride (PVDC) film+dry gauze in comparison to PVDC-film+dry gauze+aspiration tube (n=6/group). Physiological reperfusion was assessed by histology. Survival rate was determined after a 7d observation period.Only perfusion with 1% SDS resulted in an a cellular scaffold (fully translucent without histologically detectable tissue remnants, DNA<50ng/mg) with remarkable structural and ultrastructural integrity as well as preservation of main matrix proteins (immunohistochemically positive for collagen I and IV, laminin, fibronectin and elastin). Upon reperfusion, the liver turned red within sec without any leakage from the surface of the liver. 12h after reperfusion, blood cells but also some clots were visible in the portal vein, sinusoidal matrix network and central vein, suggesting physiological perfusion. Complete abdominal protection was achieved by placing a suction tube in addition to the PDVC-film+dry gauze and allowed 7d-survival of all animals.In conclusion, we successfully completed a survival model of high quality in-vivo decellularization within 2h as next step towards in-vivo liver engineering.