Aim: The development of a simplified technique for the large scale study of bone
microstructure on undefalcified bone (mineral bone histomorphometry).
Established techniques such as undecalcified bone thin sectioning, ultrathin
grinding, surface-stained block-grindings and micro-CT, electron microscopy, are
expensive, time consuming, have very high demands on equipment,
consumables, laboratory safety facilities and personell expertise.
Method: The proposed technique is based on the principles of materialography.
However, its novelty lies on the selection of user-friendly, low-cost equipment
materials in all stages of preparation (cutting, defatting, embedding, grinding,
staining, digitization). On the present thesis, all stages are described in
with many practical tips for the successful implementation: parallel thin
with a modified table band-saw arrangment, defatting with sodium hypochloride
(bleach), embedding in epoxy-resin, silicon-carbide paper grinding, von Kossa
staining, flatbed scanner digitization. For the purposes of an integrated
project, the open-source plugin “Map_BoneMicrostructure” was developed for the
platform ImageJ, for image processing and acquisition of quantitative results.
For the validation of the method, eight bone specimens were studied with the
established technique of microCT, and subsequently with the rapid
metariallographic method. The bone histomorphometry parameters BV/TV,
Tb.Th., Tb.Sp., Tb.N. were calculated by each method, and were subjected to the
statistical test of Pearson’s linear correlation. The correlation was found
significant (p<0,005) for all four parameters.
Discussion: The proposed technique is not microscope-bound, can be
implemented even on large specimens, is relatively quick, uses safe materials,
and has low demands on laboratory equipment (aprox. 1000€) and consumables
(aprox. 5€ pro slice). However, it is not suitable for the study of cellular
It was successfully validated compared with the established microCT.
Conclusion: The rapid materialographic method is an integrated research tool,
and has all prerequisites to contribute to the expansion of the study of bone
tissue, since it allows the rapid study of bone samples with low equipment and
Materialography, Bone microstructure, von Kossa staining, Undecalcified bone, Epoxy resin