Automatic decellularization of ovine aorta-derived extracellular matrix offers reduced processing and attendee times while being as effective as manual techniques

Objective: To construct an automatic decellularization platform (ADP) for preparing xenogenic extracellular matrices (ECMs), and to demonstrate that automatic decellularization for preparing xenogenic ECMs reduces processing time, requires fewer attendee hours, and is as effective as the manual gold standard preparation protocols. Materials and Methods: A soft tissue ADP was constructed and ovine aorta was harvested (n=9). Manual and automatic decellularization was performed on aortic tissue specimens and both groups were compared. The presence of acellularity was assessed with viability/cytotoxicity assays, and the presence of residual ovine DNA was determined with gel electrophoresis and spectrophotometry. Scaffold integrity was characterized with scanning electron microscopy (SEM) and uniaxial tensile testing. Results: Acellularity was confirmed with both preparation techniques and DNA concentrations measuring 540±130 and 590±270 ng/mg wet weight and the control measuring 6690±1210 ng/mg wet weight (p<0.05). SEM demonstrated no differences in the surface architecture of ECMs prepared by both techniques. Uniaxial testing demonstrated no significant differences in the incremental elastic moduli E below a stretch ratio of 2.70λ in both groups and a large reduction in E was recorded when both groups were compared with control samples above a stretch ratio of 1.7. Conclusion: Automatic decellularization of ovine aorta is as effective as gold standard manual decellularization protocols. Future research will focus on optimizing the automated decellularization technique and on upscaling protocols.