Comparison of four decontamination treatments on porcine renal decellularized extracellular matrix structure, composition, and support of human renal cortical tubular epithelium cells

J Biomater Appl. 2016 Mar;30(8):1154-67. doi: 10.1177/0885328215615760. Epub 2015 Nov 20.

Abstract

Engineering whole organs from porcine decellularized extracellular matrix and human cells may lead to a plentiful source of implantable organs. Decontaminating the porcine decellularized extracellular matrix scaffolds is an essential step prior to introducing human cells. However, decontamination of whole porcine kidneys is a major challenge because the decontamination agent or irradiation needs to diffuse deep into the structure to eliminate all microbial contamination while minimizing damage to the structure and composition of the decellularized extracellular matrix. In this study, we compared four decontamination treatments that could be applicable to whole porcine kidneys: 70% ethanol, 0.2% peracetic acid in 1 M NaCl, 0.2% peracetic acid in 4% ethanol, and gamma (γ)-irradiation. Porcine kidneys were decellularized by perfusion of 0.5% (w/v) aqueous solution of sodium dodecyl sulfate and the four decontamination treatments were optimized using segments (n = 60) of renal tissue to ensure a consistent comparison. Although all four methods were successful in decontamination, γ-irradiation was very damaging to collagen fibers and glycosaminoglycans, leading to less proliferation of human renal cortical tubular epithelium cells within the porcine decellularized extracellular matrix. The effectiveness of the other three optimized solution treatments were then all confirmed using whole decellularized porcine kidneys (n = 3). An aqueous solution of 0.2% peracetic acid in 1 M NaCl was determined to be the best method for decontamination of porcine decellularized extracellular matrix.

Keywords: Decontamination; decellularization; gamma-irradiation; kidney; peracetic acid; renal tissue.

Publication types

  • Comparative Study

MeSH terms

  • Animals
  • Cell Adhesion
  • Cell Line
  • Cell Proliferation
  • Decontamination / methods*
  • Ethanol / chemistry
  • Extracellular Matrix / chemistry*
  • Extracellular Matrix / ultrastructure
  • Gamma Rays
  • Humans
  • Kidney / chemistry*
  • Kidney / cytology
  • Kidney / radiation effects*
  • Kidney / ultrastructure
  • Peracetic Acid / chemistry
  • Swine
  • Tissue Engineering / methods
  • Tissue Scaffolds / chemistry*
  • Urothelium / cytology

Substances

  • Ethanol
  • Peracetic Acid