Toxic influence of detergents on human mesenchimal stromal cells during graft repopulation

Detergents are effective decellularization agents in order to obtain cell-free matrixes. However, the toxicity of residual detergent prevents the adhesion of cells to the matrix and the realization of their functions. In this study, the degree of influence of residual detergents on mesenchymal stromal cells after a series of washes was investigated. The human pulmonary valve were treated in three ways: 1% solution of sodium dodecyl sulfate, 1% solution of sodium deoxycholate and a combination of these solutions with the reduction of the concentration to 0.5%. After each of 8 steps washing buffer were collected. We investigated influence of residual detergents on cytotoxicity and effect on the metabolic activity of MSCs. Histologic analysis showed sufficient tissue decellularization in all groups. Matrixes were washed by successive incubations with 200 ml of phosphate buffer. Detergent concentrations in all samples ceased to be toxic only to the 4-cycle wash, and had no effect on the metabolism and viability of human MSCs. After 4-8 washing cycles decellularized pulmonary trunk fragments were successfully settled by mesenchymal stromal cells. Thus, a few successive washing fabrics pulmonary valve after decellularization reduce residual detergent to a safe level and lead to successful repopulation of acellular scaffolds with MSCs.