Rapid nodule evaluation computer-aided image analysis procedure for bone nodule quantification

Using bone cell cultures, the effects of drugs on cell activities such as proliferation, differentiation, matrix formation, and mineralization can be explored. To quantify these parameters accurately and quickly, a kinetic reproducible computed image analysis procedure of culture dishes is proposed which could be conjointly used with biochemical analysis of the medium. In the present article, different mathematical procedures coupled either with or without histochemical staining are investigated and analyzed. Using serial cross sections and microradiographies of bone nodules, we demonstrated that the gray-level parameter is well correlated with bone mass and/or the mineralization status of the nodules. The procedure selected is a multistep procedure called rapid nodule evaluation (RNE), which uses a binary reconstruction program with different thresholds. To challenge this RNE procedure with the classical Von Kossa staining and quantification procedure, we cultured the cells in the presence of 10 nmol/L dexamethasone and compared the results using the two procedures. The RNE procedure appeared to be accurate and reproducible, and also has the advantage of speed and dynamic analysis over the classical Von Kossa quantification procedure.