Evaluation of supercritical CO2 extraction protocols in a model of ovine
aortic root decellularization
Abstract
One of the leading trends in the modern tissue engineering is the
development of new effective methods of decellularization aimed at the
removal of cellular components from donor tissue reducing its
immunogenicity and the risk of rejection. Supercritical CO2 (scCO2)
extraction can significantly improve the outcome of decellularization,
reduce contamination and time costs. The resulting products can serve as
personalized tools for tissue-engineering therapy of various somatic
pathologies. However, decellularization of complex 3D structures, such
as the aortic root, requires optimization of the parameters, including
preconditioning medium composition, type of co-solvent, values of
pressure and temperature insight the scCO2 reactor, etc. In our work,
using an ovine aortic root model, we performed a comparative analysis of
the efficiency of decellularization using approaches based on various
combinations of these parameters. The protocols were based on
combinations of treatment in alkaline, ethanol or detergent solutions
with scCO2 extraction at different modes. Based on a histological
analysis, we have selected an optimal protocol for the decellularization
of ovine aortic root employing preconditioning in a detergent solution.
The positive effects of scCO2 on the decellularization extent,
cytotoxicity and histoarchitecture of the tissue were demonstrated.