Decellularization of human donor aortic and pulmonary valved conduits using low concentration sodium dodecyl sulfate

The clinical use of decellularised cardiac valve allografts is increasing. Long term data will be required to determine whether they outperform conventional cryopreserved allografts. Valves decellularised using different processes may show varied long-term outcomes. It is therefore important to understand the effects of specific decellularisation technologies on the characteristics of donor heart valves. Human cryopreserved aortic and pulmonary valved conduits were decellularised using hypotonic buffer, 0.1% (w/v) SDS and nuclease digestion. The decellularised tissues were compared to cellular cryopreserved valve tissues using histology, immunohistochemistry, quantitation of total DNA, collagen and glycosaminoglycan content, in vitro cytotoxicity assays, uniaxial tensile testing and subcutaneous implantation in mice. The decellularised tissues showed no histological evidence of cells or cell remnants and over 97% DNA removal in all regions (arterial wall, muscle, leaflet and junction). The decellularised tissues retained collagen IV and von Willebrand factor staining with some loss of fibronectin, laminin and chondroitin sulphate staining. There was an absence of MHC Class I staining in decellularised pulmonary valve tissues, with only residual staining in isolated areas of decellularised aortic valve tissues. The collagen content of the tissues was not decreased following decellularisation however the glycosaminoglycan content was reduced. Only moderate changes in the maximum load to failure of the tissues were recorded post-decellularisation. The decellularised tissues were non-cytotoxic in vitro, and were biocompatible in vivo in a mouse subcutaneous implant model. The decellularisation process will now be translated into a GMP compatible process for donor cryopreserved valves with a view to future clinical use.