Magnesium Presence Prevents Removal of Antigenic Nuclear-Associated Proteins from Bovine Pericardium for Heart Valve Engineering.
Current heart valve prostheses are associated with significant complications, including aggressive immune response, limited valve life expectancy, and inability to grow in juvenile patients. Animal derived "tissue" valves undergo glutaraldehyde fixation to mask tissue antigenicity; however, chronic immunological responses and associated calcification still commonly occur. A heart valve formed from an unfixed bovine pericardium (BP) extracellular matrix (ECM) scaffold, in which antigenic burden has been eliminated or significantly reduced, has potential to overcome deficiencies of current bioprostheses. Decellularization and antigen removal methods frequently use sequential solutions extrapolated from analytical chemistry approaches to promote solubility and removal of tissue components from resultant ECM scaffolds. However, the extent to which such prefractionation strategies may inhibit removal of antigenic tissue components has not been explored. We hypothesize that presence of magnesium in prefractionation steps causes DNA precipitation and reduces removal of nuclear-associated antigenic proteins. Keeping all variables consistent bar the addition or absence of magnesium (2 mM magnesium chloride hexahydrate), residual BP ECM scaffold antigenicity and removed antigenicity were assessed, along with residual and removed DNA content, ECM morphology, scaffold composition, and recellularization potential. Furthermore, we used proteomic methods to determine the mechanism by which magnesium presence or absence affects scaffold residual antigenicity. This study demonstrates that absence of magnesium from antigen removal solutions enhances solubility and subsequent removal of antigenic nuclear-associated proteins from BP. We therefore conclude that the primary mechanism of action for magnesium removal during antigen removal processes is avoidance of DNA precipitation, facilitating solubilization and removal of nuclear-associated antigenic proteins. Future studies are necessary to further facilitate solubility and removal of nuclear-associated antigenic proteins from xenogeneic ECM scaffolds, in addition to an in vivo assessing of the material.
1 Department of Veterinary Medicine, Medicine and Epidemiology, University of California , Davis, Davis, California.|2 Department of Cardiovascular Diseases, Mayo Clinic , Rochester, Minnesota.
Ailsa J
AJ
Dalgliesh
1 Department of Veterinary Medicine, Medicine and Epidemiology, University of California , Davis, Davis, California.
Zhi Zhao
ZZ
Liu
1 Department of Veterinary Medicine, Medicine and Epidemiology, University of California , Davis, Davis, California.|2 Department of Cardiovascular Diseases, Mayo Clinic , Rochester, Minnesota.
Leigh G
LG
Griffiths
D000818:Animals; D000941:Antigens / Q000737:chemistry*; D002417:Cattle; D002467:Cell Nucleus / Q000737:chemistry*; D005109:Extracellular Matrix / Q000737:chemistry*; D006351:Heart Valves; D008274:Magnesium / Q000737:chemistry*; D010496:Pericardium / Q000737:chemistry*; D023822:Tissue Engineering
D000941:Antigens; D008274:Magnesium
antigen removal; decellularization; extracellular matrix; heart valve; tissue engineering; xenogeneic scaffold