Creating a Tissue-Specific Microdispersed Matrix from a Decellularized Porcine Liver

The main problem with decellularization of liver tissue as a tissue-specific matrix/scaffold in liver bioengineered structures is the need to maximize the preservation of the original three-dimensional structure of the tissue and the main components of its extracellular matrix (ECM) while removing cells and genetic material. The attempts to use the existing protocols for the decellularization of other tissues and organs have been unsuccessful. The aim of the work is to develop a method for creation of a tissue-specific microdispersed matrix from decellularized porcine liver (TMM DLp). The protocol for decellularization of porcine liver (Lp) fragments has been developed on the basis of the complex application of chemical (sodium dodecyl sulfate and Triton X100), biochemical (DNase I), and physical (supercritical CO2) methods for treatment of the initial tissue. As a result of the found optimal conditions for decellularization of Lp with subsequent cryomicronization of decellularized DLp, an injectable form of the microdispersed tissue-specific matrix was obtained, which represents decellularized porcine liver microparticles with the size of 100-200 mu m with the residual amount of DNA no more than 10 +/- 1.5 ng/mg (less than 1.0%), with the preservation of the microstructure and basic composition of the liver ECM. According to the in vitro assessment, biocompatible properties of tissue-specific matrix samples meet the criteria of biological safety for cytotoxicity and hemolytic activity.