Получение микродисперсного тканеспецифического децеллюляризованного матрикса из суставного хряща свиньи

Abstract

A method for obtaining a microdispersed tissue-specific matrix from decellularized porcine articular cartilage while maintaining morphological and functional properties of the extracellular matrix with no signs of cytotoxicity was developed. The cartilage particle size distribution in suspension after cryogenic grinding was determined using laser diffraction analysis. The range of sizes of the obtained cartilage microparticles suggests the possibility of their administration by injection (<220 μm). The combination of stages, including three freeze/thaw cycles (–196/37°C) followed by treatment with solutions of surface-active substances (surfactants), sodium dodecyl sulfate and Triton X-100, and DNase resulted in the complete removal of non-decellularized microparticles. The residual DNA content was 9.11 ± 1.13 ng/mg of tissue. The effectiveness of surfactant washing was assessed by the cytotoxicity of the matrix on the culture of human adipose derived mesenchymal stromal cells (hADSCs). To assess the hemocompatibility of the obtained samples, their hemolytic activity was studied in vitro. The adhesion and proliferation of hADSCs on the matrix surface were studied on day 21 of cultivation. The matrix did not exhibit hemolytic activity and cytotoxicity with respect to hADSCs. hADSCs in the samples were actively proliferating on the matrix surface. The biocompatibility and hemocompatibility of the obtained matrix in vitro indicate its potential for application in cartilage regenerative medicine.