Comparison of SB-SDS and other decellularization methods for the acellular nerve graft: Biological evaluation and nerve repair in vitro and in vivo

Abstract

We aimed to compare the performance of acellular nerves prepared by different decellularization methods, screening out the optimal decellularization protocol, repairing the sciatic nerve defects in rats by the allogeneic transplantation, and evaluating the effect of regenerative nerve on the function reconstruction. The Sondell, SB-SDS, TnBP, and the high/low permeation methods were used to decellularize donor nerves. Nerves without any treatment were as the control group. The histological results were evaluated by HE staining and toluidine blue (TB) staining. The proliferation activity of L929 cells was detected by CCK-8 assay. The adhesion of Schwann cells was observed and quantified by SEM. Balb/c mice were used to evaluate the cellular and humoral immunogenicity of the nerve scaffolds. The rat sciatic nerve defect model was applied to observe the repair effect of acellular nerve scaffold in vivo. To SB-SDS group, it remained the original state of the nerves, with no observed nucleus and axons, the neurotoxicity grade detected by CCK-8 being almost 0, and it kept the largest number of Schwann cells adhered to the acellular nerve and the better morphology. Further, it showed that the selected SB-SDS rats acellular nerve scaffold could promote the nerve repair of the rats by HE staining and TB staining. We could conclude that the acellular nerve matrix prepared by the SB-SDS method effectively removes the cellular components in the nerve tissue and retains the main components of the extracellular matrix of the nerve tissue, whose rats decellularized nerve scaffold could promote the sciatic nerve repair better.

Keywords: SB-SDS; acellular nerve; cell compatibility; cytotoxic grading; decellularization; immunological evaluation; nerve graft; nerve injury.