A novel method for constructing an acellular 3D biomatrix from bovine spinal cord for neural tissue engineering applications

Abstract

In this study, we aimed at generating 3‐dimensional (3D) decellularized bovine spinal cord extracellular matrix‐based scaffolds (3D‐dCBS) for neural tissue engineering applications. Within this scope, bovine spinal cord tissue pieces were homogenized in 0.1 M NaOH and this viscous mixture was molded to attain 3D bioscaffolds. After resultant bioscaffolds were chemically crosslinked, the decellularization process was conducted with detergent, buffer, and enzyme solutions. Nuclear remnants in the native tissue and 3D‐dCBS were determined with DNA content analysis and agarose gel electrophoresis. Afterward, 3D‐dCBS were biochemically characterized in depth via glycosaminoglycan (GAG) content, hydroxyproline (HYP) assay, and sodium dodecyl sulfate‐polyacrylamide gel electrophoresis (SDS‐PAGE). Cellular survival of human adipose‐derived mesenchymal stem cells (hAMSCs) on the 3D‐dCBS for 3rd, 7th, and 10th days was assessed via MTT assay. Scaffold and cell/scaffold constructs were also evaluated with scanning electron microscopy and histochemical studies. DNA contents for native and 3D‐dCBS were respectively found to be 520.76 ± 18.11 and 28.80 ± 0.20 ng/mg dry weight (n = 3), indicating a successful decellularization process. GAG content, HYP assay, and SDS‐PAGE results proved that the extracellular matrix was substantially preserved during the decellularization process. In conclusion, it is believed that the novel decellularization method may allow fabricating 3D bioscaffolds with desired geometry from soft nervous system tissues.

Tissue-Specific Decellularization Methods: Rationale and Strategies to Achieve Regenerative Compounds

“…In this context, the usefulness of dECM-derived nerve scaffolds as implantable grafts has been assessed in in vitro and in vivo animal trials, with some promising results [ 174 ]. Some studies have shown the capacity of the dECM to promote axon growth and the regeneration of peripheral nerve connections in rats [ 69 , 172 , 175 ]. The regeneration process requires weeks, and the subjects are in need of rehabilitation in order to begin proper recovery.…”

Decellularized extracellular matrix scaffolds: Recent trends and emerging strategies in tissue engineering

“…Interestingly, dECM could be solubilized into a liquid phase, and subsequently form a hydrogel phase [ 264 ]. Such hydrogel scaffolds are widely investigated due to the advantages of their injectability and in situ polymerization, which delivers the hydrogels through a minimally invasive method and enhances astrocyte/axon interactions through direct integration with host tissue [ [264] , [265] , [266] ]. For instance, DeQuach decellularized porcine brains and turned them into a liquid form by using enzymatic digestion.…”

Human Hair Keratin Composite Scaffold: Characterisation and Biocompatibility Study on NIH 3T3 Fibroblast Cells

“…These α keratins can assemble together to form a microfibrous structure known as keratin and helps the suitable scaffold development. The extracted keratin was quantified using the Bradford colourimetric method as reported by Arsalan and co-workers [ 20 ].…”