Effect of MTA and CEM on Mineralization-Associated Gene Expression in Stem Cells Derived from Apical Papilla

Introduction: This study assessed the effect of mineral trioxide aggregate (MTA) and calcium-enriched mixture (CEM) cement on odontogenic differentiation and mineralization of stem cells. Methods and Materials: After confirmation of stemness and homogeneity of stem cells derived from apical papilla (SCAPs) using flow cytometry, the cells were exposed for 3 weeks to either osteogenic medium (OS) or CEM extract+OS (CEM+OS) or MTA extract in OS (MTA+OS) or DMEM based regular culture media (negative control). Relative expression of alkaline phosphatase (ALP), dentine sialophosphoprotein (DSPP), osteocalcin (OSC), and osterix (SP7) were measured at days 14 and 21 using RT-qPCR method. At the same time points Alizarin Red staining method was used to assess mineralization potential of SCAPS. Gene expression changes analysis were made automatically using REST® software and a P<0.05 was considered significant. Results: After 2 weeks of exposure, expression of all genes were between 3 and 52 times the expression of GADPH (all were upregulated except SP7 in the control, P<0.05). After 3 weeks, relative expressions of the genes: ALP, SP7, DSPP, and OSC were respectively 275.9, 528.3, 98.4, and 603.7 times the expression of GADPH in the control group (OS). These were respectively 17.405, 29.2, 11.8, and 6.5 in CEM+OS group, and 163.8, 119.7, 102.5, and 723.9 in MTA+OS group. All of these were confirmed as upregulated (P<0.05) except for ALP and OSC of DM+CEM group. After 2 weeks, alizarin red staining showed similar mineralized nodules in OS, MTA+OS, and CEM+OS. In third week, larger nodules were seen in MTA+OS and OS, but not in CEM+OS. Conclusion: After 2 weeks, gene expressions were almost comparable in OS, CEM+OS, and MTA+OS. After 3 weeks, OS and MTA+OS upregulated genes much greater than in 2nd week. However, upregulation in CEM+OS might not increase in 3rd week compared to those in 2nd week.Keywords: Biomaterials; Calcium-Enriched Mixture (CEM Cement); Mineral Trioxide Aggregate; Relative Gene Expression; Retrograde Root Filling Materials; Stem Cells from Apical Papill

Histologic evaluation of pulpal response to MTA and capsaicin in cats

Introduction: In direct pup capping, the exposed pulp is directly capped with a capping material to provoke a dentinal bridge formation to seal the exposed area.The aim of this study was to evaluate the pulpal response to MTA and capsaicin in cats. Methods: The sample for this experimental study consisted of 24 canines of cats, weighed approximately 3-4kg. After sedation, the teeth were cut and exposed at 3mm above cementoenamel junction, then divided into two groups: 1) direct pulp capping with MTA 2) direct pulp capping with Capsaicin. The cavities were filled with glass ionomer cement (Fuji IX). The cats were sacrificed in the first, second, and fourth weeks 4 canine teeth were extracted for the purpose of histologic analysis. 6Serial sections were cut parallel to the longitudinal axis of the canines. The sections were stained with hematoxylin & eosin, and were observed and analyzed using a light microscope. Results: In both groups treated with MTA and capsaicin, the inflammation decreased during weeks1, 2, and 4. Less inflammation was seen in MTA group but there was no statistical difference between the two groups (p=0.22). In the group treated with capsaicin, necrosis was observed in every 12 samples, but in the other group treated with MTA, only 1 pulp necrosis was seen in the first week (p=0.000). There was no significant difference in dentinal barrier formation, inflammatory response, and soft tissue changes between the two groups. Conclusions: The results of this study showed that capsaicin can decrease the severity of inflammation, but it is ineffective in dentinal barrier formation