Comparative study of the Triton X-100-sodium deoxycholate method and detergent-enzymatic digestion method for decellularization of porcine aortic valves

BACKGROUND: Valve replace - ment is the primary surgical treatment for heart valve disease. However, the clinical benefit of bi - ological valve substitutes is limited by their po - tential immunogenicity. AIM: To seek a better method of preparing bi - ological scaffolds for tissue engineering heart valves, we compared the ability of different de - cellularization procedures to remove cells and maintain the scaffold structure. MATERIALS AND METHODS: Specimens of fresh porcine aortic valve leaflets were randomly di - vided into group I (n = 16), group II (n = 16) and group III (n = 16).The valve leaflets in group I were not decellularized; group II were treated withTriton X-100 and sodium deoxycholate; and group III were treated by a detergent-enzymatic digestion method. The valve leaflets were investigated by gross exam - ination, hematoxylin-eosin staining, Masson's trichrome staining, and scanning electron mi - croscopy to observe the valve structure and the in - tegrity of collagen and elastin. The DNA content was measured to confirm the removal of cells. RESULTS: The detergent-enzymatic digestion method completely removed cells and led to valve fiber structural alterations in group III.TheTriton X- 100-sodium deoxycholate method achieved both complete decellularization and preservation of the valve fiber structure in group II. CONCLUSIONS: The detergent-enzymatic di - gestion method is a better technique for decellu - larization than the Triton X-100-sodium deoxy - cholate method, as it enables both complete de - cellularization and preservation of the valve scaf - fold structure.