Characterization of The Decellularized Male Rabbit Kidney as a Three- Dimensional Natural Scaffold for Tissue Engineering. A Histological Study

Abstract

Whole organ decellularization; a beacon of hope for a continuously growing population of end stage renal disease patients. The optimal whole kidney decellularization procedure yields a three-dimensional natural kidney extracellular matrix (ECM) scaffold, completely deprived from cellular materials that can elicit an immune response when implanted into a host. This decellularized ECM has the intrinsic spatial ability and biological cues that can promote cell adhesion, migration, proliferation and differentiation during recellularization. Patient’s own cells can be used for scaffold recellularization to create a functional immune-compatible kidney ready for transplantation. Vascular perfusion of detergents as sodium dodecyl sulfate (SDS) is the most widely used whole kidney decellularization protocol. Unfortunately, both SDS and perfusion pressure have a significant detrimental effect on ECM ultrastructure, vasculature and matrisome proteins. Therefore, the optimal decellularization protocol must balance between complete ECM decellularization and preservation of ECM properties. The aim of this study was to present a proper decellularization protocol for male rabbit kidney and to characterize the resultant decellularized ECM by evaluation of the structural integrity of its components as a natural scaffold ready for tissue engineering. The study was conducted on ten male New Zealand White Rabbits (NZWR) of an average weight (1000- 1500 gm). Both kidneys were harvested and sorted into two groups: • Group I (Control group): included the ten right kidneys; processed immediately for examination and analysis. • Group II (Decellularization group): included the ten left kidneys; harvested carefully after cannulation and kept frozen until decellularization. At decellularization, kidneys were thawed, perfused with 0.5% sodium dodecyl sulfate (SDS) for 5-6 hours at room temperature until kidneys were completely white. These gross morphological changes were recorded and photographed. In addition, the weight of the kidneys was recorded and statistically analyzed at the start (fresh weight), after freezing/thawing and after complete decellularization. Finally, decellularized kidneys were processed for examination and analysis as those of control group.