Construction and properties detection of 3D micro-structure scaffolds base on decellularized sheep kidney before and after crosslinking

Abstract

Decellularized extracellular matrix is one form of natural material in tissue engineering. The process of dECM retains the tissue microstructure, provides good cell adhesion sites, maintains most of biological signals that promotes the survival and differentiation ability of cells. In this study, sheep kidney was decellularized followed by histochemical staining, elemental analysis and scanning electron microscopy characterizations. The dECM scaffold was prepared with different sequences of freeze drying technology, crosslinking and the water absorption, porosity, mechanical strength with subsequent thermogravimetric analysis, Infrared spectroscopy and biocompatibility tests. Our results indicated that these decellularized treatments of sheep kidney can effectively remove DNA and retain uniform pore size distribution. After crosslinking the scaffold’s water absorption decreased from 987.56 ± 40.21% to 934.39 ± 39.61%, the porosity decreased from 89.64 ± 3.2% to 85.09 ± 17.63%, and the compression modulus increased from 304.32 ± 25.43 kPa to 459.53 ± 38.92 kPa, with thermal process the percentage of weight loss decreased from 66.57% to 44.731%, in addition, the composition didn’t change significantly, crosslinking could also promote the stability. In terms of biocompatibility, the number of viable cells increased significantly with the days. In conclusion, the crosslinked decellularized sheep kidney extracellular matrix scaffold reduced water absorption and porosity slightly, but has a significant increase in mechanical properties, and presented excellent biocompatibility which are beneficial to cell adhesion, growth and differentiation.

Crosslinking strategies of decellularized extracellular matrix in tissue regeneration

“…After EDC/NHS crosslinking, the decellularized sheep kidney scaffolds' compressive modulus increased from 304.32 ± 25.43 kPa to 459.53 ± 38.92 kPa, the thermal weight loss percentage of which decreased from 66.57% to 44.731%, while the molecular structure of the decellularized scaffold did not change before and after crosslinking. 141 In addition, the crosslinked decellularized scaffolds had excellent cytocompatibility for cell adhesion, growth, and differentiation, as exemplified by the significantly increased number of viable ADSCs with the days in vitro.…”