Brief communication: Test of a method to identify double-zonal osteon in polarized light microscopy

Double-zonal osteons (DZ) have been of interest in paleopathological research because they might be linked to physiological pathology. DZ are thought to be evidence of arrested osteon formation with a brief but abrupt increase in mineralization of lamellae occurring during bone remodeling. Originally identified from microradiographs as hypermineralized rings, recent studies have identified DZ from linear polarized light microscopy (PLM). However, PLM does not guarantee the adequate detection of DZ since PLM captures bone birefringence and not hyper-mineralization. Scanning electron microscopy with backscatter electrons (BSE-SEM) allows observation of DZ by detecting differences in mineralization. The purpose of this study is to investigate whether DZ, as identified by BSE-SEM, can indeed be identified with PLM.

The sample consists of an archaeological collection of adult midshaft femurs (n = 30) from St. Matthew cemetery, Quebec City (1771-1860). DZ were identified and counted independently with PLM and BSE-SEM for the same sections. Results from both methods were compared.

Chi-square test shows that there was no significant difference between the two methods (p = 0.404). No significant bias was found on Bland-Altman analysis and Cohen's kappa shows a substantial agreement between the two methods (Κ = 0.66). PLM shows a good accuracy (sensitivity 79%, specificity 99.4%) and reliability (Positive Predictive Value: 86.71%; Negative Predictive Value: 99.45%).

These findings indicate that the two methods are interchangeable. PLM, using our proposed protocol, is reliable to accurately identify DZ. We discuss how PLM and BSE-SEM that measure different features of the bone tissue can converge on the identification of DZ.