Comparative study of four aortic allograft decellularization techniques
Abstract
Objective: to assess four aortic allograft decellularization techniques.
Materials and methods. 40 aortic allografts were decellularized using 4 techniques (n=10 in each group), depending on reagents used: 1) 1% deoxycholic acid; 2) 0,5% sodium deoxycholate, 0,5% sodium dodecylsulfate; 3) 1% sodium deoxy-cholate, 1% Triton X-100; 4) 3% Triton X-100, 0.04% EDTA. On decellularization completion, specimens were subjected to histologic (hematoxylin and eosin) and histochemic (MSB, orcein) studies; residual DNA content was assessed, as well as mechanical testing.
Results and discussion. Decellularization with 0.5% sodium deoxycholate, 0.5% sodium dodecylsulfate resulted in com-plete cell removal with good preservation of connective tissue matrix. Statistically significant reduction of DNA content in aorta (31,05 vs 2,58 ng/mg, p<0,01) and aortic valve leaflet (39,7 vs 1,36 ng/mg, p<0,05) was revealed. According to mechanical testing results, there were no significant differences between fresh, cryopreserved and decellularized allografts.
Conclusions. Complete allograft decellularization was achieved using 0.5% sodium deoxycholate, 0.5% sodium dodecy-lsulfate, with statistically significant DNA reduction and without affecting mechanical properties of the valve.
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