Reversible extinction of the morphogen in bone matrix by reduction and oxidation of disulfide bonds

Abstract

Beta-Mercaptoethanol (beta-ME) or dithiothreitol (DTT) reduction extinguishes the capacity of bone matrix gelatin to produce new bone following implantation in a muscle pouch. If the reducing solution is used in concentrations of 50 mmoles/l or less, the extinction can be partially reversed by bubbling of oxygen through the solution for one hour. Sulfhydryl group blocking reagents prevent reoxidation of reduced bone gelatin, and restoration of the bone morphogenetic property (BMP). Unspecific borohydride reduction at 37 degrees destroys bone yield irreversibly, but at 2 degrees reduction of free aldehyde groups in bone gelatin does not prevent beta-ME and DTT reversible extinction. These observations are interpreted to suggest that the disulphide linkage may be an essential part of the biologically active conformation of either a non-collagenous bone morphogenetic polypeptide firmly bound to collagen or a collagen by-product entrapped within a water insoluble gel matrix.