Decalcification of Temporal Bones With Tetrasodium Edetate

Optimization of perfusion decalcification for bones and joints in rats

“…Demineralization can be achieved quickly by immersion in strong acids (e.g., formic acid) but at the expense of proteinic epitopes. Immersion in weakly acidic chelating agents for extended period of time can also alter the cellular structures and reduce affinity for immunohistochemistry (Gussen and Donahue, 1965; Eggert and Germain, 1979; Kiviranta et al, 1980; Matthews and Mason, 1984). In this study, perfusion techniques considerably shortened the decalcification time of peripheral joints compared with immersion: 8 weeks were required for complete decalcification of knee joints of a rat weighing 200–350 g with immersion in EDTA, whereas with systemic EDTA perfusion, only 7 days were required.…”

“…Most histological studies on osseous tissues require decalcification. Although decalcification with strong acids is fast, it usually damages most cellular and subcellular epitopes (Clark, 1954; Gussen and Donahue, 1965; Bussolati, 1978; Eggert and Germain, 1979; Kirvanita et al, 1980; Matthews and Mason, 1984; Nilsson et al, 1991). To preserve these subcellular structures, hard tissue decalcification is achieved generally by immersion in calcium chelating agents such as ethylenediamine tetraacetic acid (EDTA) from several weeks to a few months (Matthews and Mason, 1984; Sano et al, 1992).…”