Table 1.
Various methods for the decellularization of heart valves.
| Decellularization method | Treatments/chemicals | General effectiveness | General effect on valve ECM | Ref. |
|---|---|---|---|---|
| Anionic detergent | SDS or sodium deoxycholate | Lack of visible cell nuclei; ~95% DNA removal | Increased areal strain and peak stretch ratio; decreased flexural stiffness; preservation of GAGs; can disrupt ECM fiber structure | 14,36–42 |
| Non-ionic detergent | Triton X-100 | Lack of visible cell nuclei | Increased areal strain and peak stretch ratio; decreased flexural stiffness; loose ECM network; histological reduction of GAG, laminin, fibronectin, and collagen | 14,39,43–45 |
| Multi-detergent | Triton X-100 + sodium cholate | ~30% DNA removal | Increased extensibility and decreased stiffness; GAG reduction; preservation of elastin and collagen components | 46,47 |
| Triton X-100 + sodium deoxycholate | Lack of visible cell nuclei; 98% DNA removal | Histologic preservation of structure and ECM components | 39,48 | |
| Osmotic shock + Triton X-100 + NLS salt + ethanol | Lack of visible cell nuclei; >97% dsDNA removal | Increased areal strain and peak stretch ratio; decreased stress relaxation; reduced GAG content | 12,49 | |
| Enzymatic | Trypsin + EDTA | Incomplete cell removal | Decreased mechanical properties; histologic tissue damage and loss of basement membrane; histologic reduction of GAG, laminin, fibronectin, and collagen | 14,40,41,43–45,50 |
| Enzymatic combinations | Trypsin + SDS | Lack of visible cell nuclei; 96% DNA removal | Reduction of GAG and α-Gal antigen; preservation of mechanical properties | 51 |
| Trypsin + sodium deoxycholate | Visible cell remnants; 98% DNA removal | Histologic disruption of ECM components | 48 | |
| Trypsin + osmotic shock + Triton X-100 | Lack of visible cells | Misalignment of collagen fibers | 41,52 | |
| Trypsin + osmotic shock | Visible cell remnants | Histologic loss of collagen; GAG reduction; decreased mechanical strength | 45 | |
| Glycol radiation | PEG + gamma irradiation | Lack of visible cell nuclei; >92% cusp DNA removal | Preserved leaflet ultrastructure; removal of α-Gal antigen | 53 |
| Osmotic shock | Hypotonic/hypertonic Tris buffer | Many visible cell remnants | Histologic reduction in MHC antigens; loss of non-collagen proteins | 44,45 |
| Sequential antigen removal | Dithiothreitol, potassium chloride, amidosulfobetaine-14 | Lack of visible cell remnants and reduced antigenicity | Preservation of Young’s modulus and ultimate tensile strength; preservation of collagen and elastin; decreased GAGs | 54–56 |
| Supercritical fluid | CO2; ethanol | Lack of visible cell nuclei; 90% phospholipid removal | Stiffening of tissue; tissue dehydration | 57 |
ECM: extracellular matrix; SDS: sodium dodecyl sulfate; GAG: glycosaminoglycan; NLS: N-lauroylsarcosine sodium salt; dsDNA: double-stranded DNA; EDTA: ethylenediaminetetraacetic acid; α-Gal: galactose-α(1,2)-galactose; PEG: polyethylene glycol; MHC: major histocompatibility complex.
The general effectiveness and general effect on ECM are overall observations and individual results may vary based on the protocol or tissue used.