RESUMO
Objective:To study the correlation between the residual amounts of commonly used tissue decellularization reagents like sodium dodecyl sulfate (SDS), Triton X-100, sulfobetaine 10 (SB-10) and cytotoxicity, and to find the safety threshold for controlling the residual of the reagent.Methods:SDS, Triton X-100, SB-10 solutions of different concentrations were prepared, and the cytotoxicity of solutions of different concentrations was detected by L-929 cells and MC3T3-E1 cells according to the method of GB/T 16886.5. The mass concentration range of the above three decellularization reagents was refined, and the threshold for the residual non-cytotoxicity of a single reagent was determined. Mix the three reagents in pair or three blends, and determine the threshold of non-cytotoxicity after the reagents are mixed by cytotoxicity test.Results:SDS, Triton X-100 and SB-10 caused L-929 cells to produce cytotoxic concentration values of 4×10 -3, 2×10 -2 and 6×10 -1 mg/ml, and MC3T3-E1 cells 6×10 -3, 4×10 -2 and 2 mg/ml. The toxicity trend of the three reagents is SDS>Triton X-100>SB-10. When three reagents at critical concentrations are mixed in pairs or three, they have no cytotoxicity to L-929 cells and MC3T3-E1 cells. Conclusions:When using the above three decellularization reagents to prepare decellularized scaffolds, a variety of reagent cleaning procedures should be used to efficiently clean the tissues so that the residual amount of reagents is below the critical value to ensure the biological safety of the decellularized scaffolds.
