Staining intensity of individual osteons correlated with elastic properties and degrees of mineralization

Abstract

Surface staining is widely used for histological studies involving undecalcified thick bone sections. Individual osteons, particularly newly formed ones stained with toluidine blue (TB), show various color intensities. We studied the correlations between TB color intensity and the differences in stiffness and degree of mineralization of individual osteons in undecalcified histological sections of goat tibial diaphysis, measured by scanning acoustic microscopy (SAM) and contact microradiography (CMR), respectively. Results showed that all three measurements correlated significantly with each other (r = 0.567 - 0.786; all P < or = 0.01). The TB surface staining intensity of individual osteons correlated better with the reflection coefficient (stiffness index) measured by SAM (r = 0.713) than with the aluminum step-wedge equivalent thickness measured on CMR micrographs (r = 0.567). The aluminum step-wedge equivalent thickness of individual osteons on CMR correlated slightly better with the SAM reflection coefficient (r = 0.786) than with the TB surface staining intensity (r = 0.713). The results of this study suggest that TB surface staining may be used as a simple method for indicating differences in stiffness and degree of mineralization in individual osteons in comparative histological studies.

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“…8a) [110]. The method was controlled by SAM and microradiography and the staining intensity negatively correlates with the degree of mineralization [111]. The staining does not work on thin microtome sections, and it is likely that the large amount of the poly-methylmethacrylate remaining in polished sections limits the access of the stain since decalcified bone embedded similarly is uniformly and deeply stained.…”

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