Detection of Experimentally Produced Occult Microfractures at the Bone-Cartilage Interface in Decalcified Sections

We compared three histological preparation methods to detect experimentally produced occult microfractures in decalcified human patellae: a paraffin tape-transfer technique, a paraffin slab-cut method, and a paraffin method with methyl salicylate as the clearing agent. Microfractures were observed at the bone-cartilage interface and were oriented either parallel or perpendicular to the tidemark. Both types of microfractures were documented with each preparation method. The slab-cut method was time-consuming, but the section thickness allowed detailed analysis of the architecture of microcracks as they passed into the depth of the section. The methyl salicylate method was efficient and produced thin, serial sections with good morphological detail and minimal cutting artifact. Reliable histological data were also derived from the tape-transfer technique, but this method was inconsistent. The methods summarized here for processing decalcified human joint tissues provide a basis for future orthopaedic studies investigating occult microfractures at the bone-cartilage interface.