Optimization of perfusion decalcification for bones and joints in rats

Decalcification of osseous tissues by perfusion of decalcifying solution in to the vascular system has never been applied to the study of peripheral jo ints. To optimize perfusion methods, rats were decalcified by direct immers ion or by one of two perfusion techniques: 1) systemic perfusion circulatin g the decalcifying solution from the ascending aorta; and 2) regional perfu sion circulating the solution to the lower extremities from the abdominal a orta. The process of decalcification was monitored by serial radiographic e xaminations. After decalcification, bone and joint samples were stained for histochemistry and immunohistochemistry. With systemic perfusion, the deca lcification time, dependent on weight, was markedly reduced compared to imm ersion. Regional perfusion decalcification was faster than all other method s studied. Microstructural preservation was comparable and immunostaining q uality often improved. Applications of this work will improve the study of basic skeletal and articular problems. Anat Rec 260:222-227, 2000. (C) 2000 Wiley-Liss, Inc.