Optimization of decellularization and biocompatibility assessment of laser-perforated vascular matrices

Background: To enable the adaptation of biological materials intended for the prosthetics of various parts of the cardiovascular system to the physiological conditions of the body, research aimed at a comparative analysis of decellularization protocols for creating an optimal scaffold, as well as improving its properties, in particular through surface modification, is of current importance. Objective: The study aims to optimize the protocol on decellularization of the porcine pulmonary trunk wall by comparative analysis of detergent compositions followed by assessement of the effect of laser microperforation on the biocompatibility and cellular repopulation of the decellularized matrix in vitro and in vivo. Methods: The first stage of the work was aimed at selecting the incubation time for porcine pulmonary trunk fragments in solutions with various concentrations of detergents, followed by tensometric tests to determine the optimal combination of conditions. Further, the decellularized material was perforated using laser processing. Some of the resulting samples were repopulated with fibroblast cells for 7 days, while other samples were subcutaneously implanted into the paravertebral region of rats for 30 days. Histological studies of the biomaterials were conducted at each stage. Results: The optimal decellularization protocol for the porcine pulmonary trunk wall was determined to be its treatment with a 0.5 % sodium dodecyl sulfate solution in combination with 0.5 % sodium deoxycholate, which enables to produce an acellular matrix with preserved ultrastructure within 16 hours. Culturing mouse fibroblasts on the surface of laser-treated samples proved the sustention of cell viability. The investigation of the cellular composition of explants retrieved 30 days after the experimental surgery showed that laser perforation preserves the material’s biocompatibility. Conclusion: The conducted research resulted in the development and validation of an effective decellularization protocol for obtaining an acellular pulmonary trunk wall matrix with preserved extracellular structure. Moreover, laser perforation of the matrices does not compromise their biocompatibility and enhances their integrative potential by expanding the contact surface area.