A study of biocompatibility of acellular matrices of primate lungs and the potential for their application as tissue-engineering constructions

Abstract

The aim of the study was to evaluate biocompatibility of acellular matrices on the model of primate lungs, which is closely approximated to human lungs in terms of anatomic-morphological characteristics, for the purpose of creating tissue engineering constructions. Materials and Methods. Acellular matrix was obtained from detergent-enzymatic decellularization of primate lungs according to a modified protocol, where exposition time and detergent concentration were increased. The quality of the obtained scaffold was morphologically evaluated after staining with hematoxylin and eosin and DAPI fluorophore, and by testing the assay for residual DNA. The change in the composition of decellularized extracellular matrix was quantitatively evaluated during the registration of the square area of positive staining after immunohistochemical testing and during staining with alcian blue. Biocompatibility of scaffolds was tested with XTT assay after 48 and 72 h of static recellularization with a consequent quantitative determination. As the main cellular resource for colonization of the scaffolds, we used multipotent mesenchymal stem cells of primates whose cellular affiliation was verified by immunophenotyping and targeted differentiation. Results. Morphological testing did not reveal any considerable structural damage of the scaffolds, residual DNA was 19.2%. Quantitation of the square area of positive staining revealed a statistically significant decrease in the amount of type I collagen and the vascular endothelium growth factor (VEGF) after decellularization. Quantitative changes in extracellular matrix were registered, however they do not impair biocompatibility of the scaffolds and do not affect cell activity. Conclusion. The obtained scaffolds do not have cytotoxic properties, they contribute to adhesion, growth and metabolic activity of cells, which allows us to consider acellular matrices to be a potential for creating tissue engineering constructions of lungs after additional enhancement of the protein composition.