Figure 2

Thickened cervical cementum formed by the inactivation of Enpp1 is enriched with non-collagenous proteins. (a) The surface structure of the distal root of the mandibular first molar at P28 was observed by SEM analysis (Most left). Tissue sections of the mandibular first molar from Enpp1^asj and the control mice at P28 were stained with H-E (Second left). Ectopic cementum (EC) was intensively stained with hematoxylin. Molecular changes of Npp1, Bsp, Dmp1, and Opn in the cervical cementum were detected by IHC staining with tissue sections of the mandibular first molar at P28 as indicated. Black arrow indicates the thickened cervical cementum of Enpp1^asj mice with the IHC pattern that was Npp1-negative and strongly positive with Bsp, Dmp1, and Opn. D, dentin; PDL, periodontal ligament; C, cementum; EC, ectopic cementum. Scale bars are indicated. (b) Cementum thickness was measured using H-E-stained tissue sections from WT and Enpp1^asj mice at indicated ages (n = 10). (c) Transcript levels of Bsp, Dmp1, and Opn were analyzed by real-time qPCR (n = 3). RNA was isolated from OCCM-30 cells with shEnpp1 and shNC harvested before and after differentiation with 10 mM β-GP treatment for 4 days. Significance was assigned with p-values as indicated in the graph.