Development of a porcine renal extracellular matrix scaffold as a platform for kidney regeneration

Abstract

Acellular scaffolds, possessing an intact three-dimensional extracellular matrix (ECM) architecture and biochemical components, are promising for regeneration of complex organs, such as the kidney. We have successfully developed a porcine renal acellular scaffold and analyzed its physical/biochemical characteristics, biocompatibility, and kidney reconstructive potential. Segmented porcine kidney cortexes were treated with either 1% (v/v) Triton X-100 (Triton) or sodium dodecyl sulfate (SDS). Scanning electron microscopy showed both treatments preserved native tissue architecture, including porosity and composition. Swelling behavior was higher in the Triton-treated compared with the SDS-treated scaffold. Maximum compressive strength was lower in the Triton-treated compared with the SDS-treated scaffold. Attenuated total reflective-infrared spectroscopy showed the presence of amide II (-NH) in both scaffolds. Furthermore, richer ECM protein and growth factor contents were observed in the Triton-treated compared with SDS-treated scaffold. Primary human kidney cell adherence, viability, and proliferation were enhanced on the Triton-treated scaffold compared with SDS-treated scaffold. Following murine in vivo implantation, tumorigenecity was absent for both scaffolds after 8 weeks and in the Triton-treated scaffold only, glomeruli-like structure formation and neovascularity were observed. We identified 1% Triton X-100 as a more suitable decellularizing agent for porcine renal ECM scaffolds prior to kidney regeneration.

Renal Extracellular Matrix Scaffolds From Discarded Kidneys Maintain Glomerular Morphometry and Vascular Resilience and Retains Critical Growth Factors

“…In particular, extracellular matrix (ECM) scaffolds obtained through the detergent-based decellularization of multiple species are used as a template for the seeding of kidney-specific cells or progenitor cells, in an attempt to regenerate the parenchymal compartment, as well as of endothelial cells or progenitor cells aiming at the full regeneration of the endothelium. Since the very first report by Ross et al 3 on the production of bioactive ECM scaffolds from rodent kidneys, several studies have followed [4][5][6][7][8][9][10][11][12] and have provided evidence that renal ECM scaffolds can be successfully and consistently produced from virtually all species including humans, 9,13 are completely acellular and virtually nonimmunogenic, maintain their architecture and essential molecular composition, lack cell membrane molecules, are able to determine cell phenotype and induce genes of renal development, possess remarkable angiogenic properties as demonstrated by the ability to induce vessel formation in the chorioallantoic membrane, are biocompatible in vitro and in vivo, and, when repopulated with renal cells, are able to show some function. Moreover, when acellular porcine renal ECM scaffolds are implanted in pigs, the framework of the innate vasculature remains well preserved and is able to well-sustain physiologic blood pressure.…”

Advances in the Knowledge about Kidney Decellularization and Repopulation

“…On the other hand, decellularization with SDS could sufficiently remove cells while preserving the ECM (Matthiesen et al, 2007; Tanemoto et al, 2008; Nakayama et al, 2010, 2011, 2013; Ross et al, 2012; Sullivan et al, 2012; Caralt et al, 2015). Choi et al (2015), however, obtained opposite results regarding the comparison between the effects of SDS and Triton X-100 on porcine kidneys. They observed that a richer ECM protein and growth factor content were present in the Triton-treated scaffold than in the SDS-treated scaffold; additionally, cell viability, adherence, and proliferation were higher with the Triton-treated scaffold than with the SDS-treated scaffold (Choi et al, 2015).…”

“…Choi et al (2015), however, obtained opposite results regarding the comparison between the effects of SDS and Triton X-100 on porcine kidneys. They observed that a richer ECM protein and growth factor content were present in the Triton-treated scaffold than in the SDS-treated scaffold; additionally, cell viability, adherence, and proliferation were higher with the Triton-treated scaffold than with the SDS-treated scaffold (Choi et al, 2015). Furthermore, other researchers have demonstrated that the combination of the two agents produces better results (Sugawara et al, 2011).…”

A critical review of current progress in 3D kidney biomanufacturing: advances, challenges, and recommendations

“…It is not only the inclusion of factors, but also the distribution that are important for cell homing and differentiation [183]. Potent growth factors including fibroblast growth factor (FGF), hepatocyte growth factor (HGF), vascular endothelial growth factor (VEGF), and interleukin-8 (IL-8) are all important for cellular fate and growth [57,185]. Insulin-like growth factor (IGF), HGF, and FGF-9 are all crucial for kidney-specific gene expression [25,172,186].…”