Surfactant-free preparation of an ostrich carotid artery scaffold using liquefied dimethyl ether and DNase

Instead of combining the surfactant sodium dodecyl sulphate and DNase, a combination of liquefied dimethyl ether (DME) and DNase was used to decellularise the scaffold of ostrich carotid artery tissue. Firstly, lipids were extracted from ostrich carotid artery tissue using liquefied DME at 25 °C and a pressure of 0.59 MPa. After DME extraction, the ostrich carotid artery tissue was collected from the extraction column, and the DME remaining in the tissue was evaporated at atmospheric pressure and temperature. DNA fragmentation by DNase was then carried out using a method almost identical to the conventional method. Finally, the tissue was washed to remove fragmented DNA. The DNA was completely fragmented to a size of less than 100 bp after 1 day of DNase treatment. The residual DNA had a concentration of 28 ng/mg dry weight after 7 days of treatment with DNase. Haematoxylin and eosin staining showed that most of the cell nuclei were removed from the aortic tissue. These results indicate that the combination of liquefied DME extraction and DNase treatment eliminates the need for surfactant treatment in ostrich carotid artery tissue decellularisation. Although previous decellularisation studies have focused on porcine tissue, we herein show the potential of ostrich tissue as an alternative to alleviate religious concerns.