Qualitative and quantitative assessment of intratubular dentin formation in human natural carious lesions

It was the aim of this study to investigate the scanning electron microscopy (SEM) features and the element content of calcium (Ca) phosphorus (P), magnesium (Mg), oxygen (O), and carbon (C) of sound dentin, demineralized dentin, secondary dentin, intratubular and peritubular de novo dentin formation due to caries progression and to compare the element content with chemically pure hydroxyapatite. Eighteen extracted teeth with deep dentin carious lesions were embedded in Technovit 9100 (Kulzer), and serial sections of 80 microm thickness were made. These sections were then investigated with polarized light microscopy to identify the lesions. Two sections of each lesion were then coated with carbon and studied with a scanning electron microscope. Of the 18 teeth, 8 showed intratubular dentin formation. The element content was measured using energy dispersive X-ray analysis (EDX). About 75% of all involved dentin tubules showed intratubular de novo dentin formation. The Ca/P ratio in sound dentin, demineralized dentin, peritubular dentin, and secondary dentin was within the range of hydroxyapatite, whereas in intratubular dentin the Ca/P ratio was different than that of hydroxyapatite. The element content for Ca was statistically highly significantly different (P < 0.01) among sound dentin/intratubular dentin, sound dentin/peritubular dentin, and sound dentin/ secondary dentin, but not between sound dentin/ demineralized dentin and sound dentin/hydroxyapatite. For the other measured elements varying statistical differences were found. Our results indicate that intratubular dentin does not occlude the dentin tubules completely and mineralizes different than that of sound dentin and may therefore not be true hydroxyapatite.