Solubility of a Bone-inducing Substance from a Murine Osteosarcoma

Abstract

A bone-inducing substance (osteogenic factor) from a murine osteosarcoma was found to be soluble in 1% sodium lauryl sulfate (SDS), 1% deoxycholate, 1% Triton X-100, and 1% Nonidet P-40. A precipitate formed on removal of the detergents by dialysis against phosphate buffer, and this precipitate induced ectopic bone formation when implanted into allogeneic mice. The insoluble residue left after extraction with SDS or deoxycholate did not evoke new bone formation, indicating that the substance was solubilized completely. The bone-inducing substance was also partially solubilized with weak acids (pH, 2.6–3.0) but not with acidic solutions of lower or higher pH. These findings indicate that the solubility of the substance depends on the hydrogen ion concentration of the solution. The substance was not solubilized with EDTA or 6 M urea.