The effect of tissue condition and applied load on Vickers hardness of human trabecular bone

Hardness of trabecular human bone, evaluated by microindentation testing, has generally been measured on embedded tissues. It was known that this was not ideal but it had been preferred to other conditions (e.g. wet or dehydrated) as the trabeculae could withstand the applied load and the measurements were reliable. The aim of this study was to investigate if the tissue condition of the specimen and the applied load would alter the hardness values measured by Vickers microindentation. Vickers hardness values of human trabecular bone from the femoral head, prepared in three different ways (wet, dry and embedded) and tested with two different loads (50 and 25 gf), were measured. No significant difference was found between the two different loads. However, in several cases the 50 gf indentations had to be redone because they were too large or the trabecula broke locally. Even if the outlines of the indentations on wet bone were slightly less marked than the ones done on dehydrated or embedded bone, it was possible to measure the hardness. Significant differences of Vickers hardness values were found between the three preparations: the hardness increased passing from wet to dried (10%) and from wet to embedded (35%). Whereas the variation coefficient of the three tissue conditions were comparable. In conclusion, it is recommended to test human trabecular bone in a wet condition as it better represents the in vivo condition. Furthermore the use of a 25 gf load is suggested, allowing hardness measurements on almost all trabeculae without breaking them.