“…Physical methods can either rely on snap freezing (Jackson et al, 1988;Roberts et al, 1991), mechanical force (Freytes et al, 2004) or mechanical agitation (Schenke-Layland et al, 2003), whereas enzymatic protocols employ nucleases, calcium chelating agents or protease digestion (Teebken et al, 2000;Bader et al, 1998;McFetridge et al, 2004;Gamba et al, 2002). Regarding physical decellularization processes, sonication, based on the use of ultrasounds to disrupt the cell membrane, has been investigated.…”