Development of renal extracellular matrix (ECM) scaffold for kidney regeneration

Abstract

The innate extracellular matrix (ECM) scaffolds can be a promising scaffold for regeneration of complex organ such as heart, liver and kidney. They possess intact 3-dimentional architecture and biochemical components that allow to access to the organ’s capillary network. In this study we have developed a porcine renal ECM scaffold and analyzed its physical and biochemical characteristics including biocompatibility for human kidney regeneration. A segmented porcine kidney cortex was obtained and treated with 1% (v/v) Triton X-100 (Triton) or sodium dodecyl sulfate (SDS) in a shaking chamber, and rinsed with distilled water. After confirmation of decellularization with H&E stain, the matrix was lyophilized and sterilized. Scanning Electron Microscope (SEM) analysis showed that both scaffolds were preserved with proper architecture including porosity for cell adhesion and composition of the renal ECM. The water uptake ability of the Triton treated scaffold was higher than that of SDS treated one. The maximum compressive strength of Triton was lower than SDS treated scaffolds and that correlates with the results of porosity and water uptake analysis. In ATR-IR analysis, both scaffolds showed a peak at 3445–3446 cm−1 and that indicates the presence of amide II (-NH). Triton treated scaffold demonstrated that there are richer contents of ECM proteins and growth factors compared to SDS treated one. When scaffolds were seeded with primary human kidney cells, Triton treated scaffold showed 2.66 times higher number of adherent cells than SDS treated one at 24 hrs post-seeding. On a CCK-8 analysis, the Triton treated scaffold showed significantly higher cell viability and proliferation rate than that of SDS treated one. Both scaffolds had no tumorigenecity for 8 weeks in vivo analysis. In conclusion, we successfully developed porcine renal ECM scaffold and confirmed that there is a great potential of porcine renal ECM scaffold to be used as human kidney regeneration. We also verified that 1% Triton X-100 is more suitable decellularizing agent than SDS regarding structural, biochemical integrity and biocompatibitilty of the scaffold. To support our findings and human application of practical regeneration, we are planning to perform in vivo experiment for kidney regeneration near future.