The expression of the PTHrP and/or PTHR1 (parathyroid hormone receptor 1) appears to be crucial to normal tooth development in both rodent and human.10, 11 and 12 Calvi et al.13 reported foetal and neonatal odontogenesis in collagen promoter-driven constitutively active PTHR1 mice, and described the consequences of the activation as odontoblastic maturation delay and formation of abnormal dentine matrix. In addition, PTH can stimulate dentine apposition in the thyroparathyroidectomized rat in a dose-dependent manner.14 Understanding PTH function Apitolisib ic50 in cells associated with

teeth formation is important for broadening our knowledge of the EPZ015666 research buy regulatory role of PTH during formation and

regeneration of all mineralized tissues. Recently we showed that during mouse incisor formation the intermittent PTH administration caused an increase of the dentine apposition rate, dentine microhardness and also the relative concentration of Ca and P in the peritubular dentine.15 Therefore, in the present study we evaluated the effect of the PTH treatment (continuous or transient) on odontoblast-like cells (MDPC-23) under the following parameters: calcium deposition, ALP, COL1, MMP-2 and BGN gene expression, ALP and MMP-2 activities. Murine odontoblast-like cells line MDPC-2316 were cultured in Dulbecco’s modified Eagle’s medium (DMEM) (Cultilab, SP, Brazil) supplemented with 10% heat-inactivated foetal bovine serum (FBS) (LGC

Biotecnologia, SP, Brazil) and penicillin (100 units/mL)/streptomycin (100 μg/mL) (GIBCO, Auckland, New York, USA) at 37 °C in an atmosphere of high humidity and 5% CO2. Initially, the cells were plated at a concentration of 2 × 105/mL in multi-well plates and cultured for 72 h until they had reached a confluent state. In order to assess whether different ways of PTH treatment could modulate the MDPC-23 response, confluent Montelukast Sodium cells were cultured in the presence of 50 ng/mL hPTH (1–34) (Sigma–Aldrich, St. Louis, MO, USA) diluted in H2O for 1 or 24 h within a 48-h incubation cycle, and then cultured without PTH for the remaining time of the each cycle. These cycles were carried out three or ten times (mineralization assay) and the analyses were performed at the end of experiment period. This intermittent treatment regimen was used to mimic the potentially anabolic effects of PTH.17 and 18 In parallel, the cells were subjected to continuous PTH exposure throughout the entire experimental period. During this experimental period the culture medium was supplemented with 2% FBS (except for mineralization assay) and changed each 48 h. Vehicle-treated for each group and untreated cultures served as controls.