Comparative proteomic analysis of the composition of decellularized extracellular matrix (dECM) and dECM-based inks as compared to the native tissue

Regenerative medicine and tissue engineering approaches based on the use of 3D-bioprinted decellularized extracellular matrix (dECM) present the advantage of a relatively biomolecule-rich matrix, which directs cell growth and differentiation in a tissue-specific manner. However, little is known about the composition changes that occur with standard processing of dECM-based inks. To characterize this process, six porcine tissues/tissue layers (artery, breast, dermis, epidermis, muscle and nerve) were independently decellularized via chemical, mechanical and enzymatic processes and the resulting dECMs formulated into biocompatible inks, to serve as source biomaterials for 3D printing. A comparative liquid chromatography–tandem mass spectrometry (LC–MS/MS)-based proteomic analysis was carried out for native tissue, decellularized and formulated ECMs, and the resulting complexity of the matrisome analyzed. A core matrisome was found to overlap in all decellularized tissues, as well as tissue-specific components that correlated with predicted functional (gene ontology-based) definitions. The proportion of collagens (mostly the α1 chains of collagen type I and III) increased in the final processing step (inks) as compared to the native ECM and dECM stages. Overall, a median of 55 matrisomal proteins (range 45-126) was detected in the dECM-derived inks. This complexity is far superior in terms of mimicking the composition of native tissue to non-dECM-based inks. Our results support the use of dECM-based inks and biomaterials in mimicking native tissue ECM complexity.