Bone histomorphometry using interactive image analysis. A methodological study with application on the human...

The aim was to develop a reproducible method for bone histomorphometry with the aid of a computerized image analysis program, and to examine the variation when assessing the total and the trabecular bone volume. Histologic sections of 18 temporomandibular joint autopsy specimens were read interactively using a cursor. The two parameters total bone volume and trabecular bone volume, of the condyle and the temporal component respectively, were estimated 2 x by 1 observer using 3 different threshold settings: an automatic, a semi-automatic and a manual technique. The threshold was based on the gray-scale distribution of the image. 2 observers read the same sections with the aid of the semi-automatic technique. The intra-observer variation expressed as coefficient of variation ranged between 1.9% and 7.1% for the different parameters, when the automatic threshold setting technique was employed, and between 2.8% and 8.7% with the semi-automatic technique. The manual technique resulted in a high intra-observer variation with a coefficient of variation between 5.2% and 19.9%. There was a systematic difference between the estimates of the 2 observers. In general, intra- and inter-observer variation was higher in the temporal component than in the condyle. The inter-section variation was moderate, the coefficient of variation ranging from 3.8% to 11.1%. The automatic and semi-automatic techniques resulted in comparable intra-observer variation, with a lower bias in the estimates of the semi-automatic technique. By letting one observer apply the semi-automatic technique, it was possible to achieve fast and reproducible analysis of the total and trabecular bone volume.