4.3 Article

Investigating the interfaces of the epiphyseal plate: An integrated approach of histochemistry, microtomography and SEM

Journal

JOURNAL OF ANATOMY
Volume -, Issue -, Pages -

Publisher

WILEY
DOI: 10.1111/joa.13924

Keywords

electron microscopy; growth plate; microtomography; ultrastructure

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In this study, we investigated the interfaces of the epiphyseal plate with over- and underlying bone segments using an integrated approach of histochemistry, microtomography, and scanning electron microscopy. The two interfaces showed significant differences, with the diaphyseal side exhibiting active calcification and slow remodeling of cartilage into bone, while the epiphyseal side showed marginal and discontinuous mineralization and a loose trabecular bone structure. The calcification process in both sides involved the formation of spheroidal bodies, which is distinct from the calcification process in bone and other tissues.
We investigated the interfaces of the epiphyseal plate with over- and underlying bone segments using an integrated approach of histochemistry, microtomography and scanning electron microscopy (SEM) to overcome the inherent limitations of sections-based techniques. Microtomography was able to provide an unobstructed, frontal view of large expanses of the two bone surfaces facing the growth plate, while SEM observation after removal of the soft matrix granted an equally unhindered access with a higher resolution. The two interfaces appeared widely dissimilar. On the diaphyseal side the hypertrophic chondrocytes were arranged in tall columns packed in a sort of compact palisade; the interposed extracellular matrix was actively calcifying into a thick mineralized crust growing towards the epiphysis. Behind the mineralization front, histochemical data revealed a number of surviving cartilage islets which were being slowly remodelled into bone. In contrast, the epiphyseal side of the cartilage consisted of a relatively quiescent reserve zone whose mineralization was marginal in amount and discontinuous in extension; the epiphyseal bone consisted of a loose trabecular meshwork, with ample vascular spaces opening directly into the non-mineralized cartilage. On both sides the calcification process took place through the formation of spheroidal bodies 1-2 mu m wide which gradually grew by apposition and coalesced into a solid mass, in a way distinctly different from that of bone and other calcified tissues.

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