Cryopreservation induces macrophage colony stimulating factor from human periodontal ligament cells in vitro.

Cryopreservation is used to protect vital periodontal ligaments during the transplantation of teeth. We investigated which gene products implicated in root resorption are upregulated in human periodontal ligament cells by cryopreservation, and whether cryopreservation affects the expression of macrophage-colony stimulating factor (M-CSF) in human periodontal ligament cells. We used customized microarrays to compare gene expression in human periodontal ligament cells cultured from teeth immediately after extraction and from cryopreserved teeth. Based on the result of these assays, we examined M-CSF expression in periodontal ligament cells from the immediately extracted tooth and cryopreserved teeth by real-time PCR, enzyme-linked immunosorbent assay (ELISA), Western blot analysis, and immunofluorescence. We also investigated whether human bone marrow cells differentiate into tartrate-resistant acid phosphatase (TRAP) positive osteoclasts when stimulated with RANKL (Receptor Activator for Nuclear Factor κ B Ligand) together with any secreted M-CSF present in the supernatants of the periodontal ligament cells cultured from the various groups of teeth. M-CSF was twofold higher in the periodontal ligament cells from the rapid freezing teeth than in those from the immediately extracted group (p < 0.05). Cryopreservation increased M-CSF expression in the periodontal ligament cells when analyzed by real time PCR, ELISA, Western blotting, and immunofluorescence (p < 0.05). TRAP positive osteoclasts were formed in response to RANKL and the secreted M-CSF present in the supernatants of all the experimental groups except negative control. These results demonstrate that cryopreservation promotes the production of M-CSF, which plays an important role in root resorption by periodontal ligament cells.

The effects of humidity and serum on the surface microhardness and morphology of five retrograde filling materials.

The purpose of this study was to compare the surface morphology and surface hardness of five materials 24 h after filling, in conditions of 100% humidity, and fetal bovine serum. The five materials were ProRoot Mineral Trioxide Aggregate (MTA), Super-EBA, Intermediate Restorative Materials (IRM), Zinc Oxide Eugenol (ZOE), and Amalgam. The microhardness of these materials was evaluated by Vickers microhardness test, and their morphologies were compared by using scanning electron microscopy (SEM). To evaluate the microhardness, the mixed five materials were measured with Vickers microhardness test. Differences between the experimental groups were analyzed by two-way ANOVA and Duncan's multiple comparison tests. All analyses were performed using the Statistical Package for the Social Sciences (SPSS Inc., Chicago, IL). For the microstructural morphological evaluation, the cross cut and root-end cavity prepared surfaces followed by retrograde filling with five different materials were observed under a Scanning Electron Microscope (Steroscan 440; Leica, Cambridge, England) at ×500. To summarize, Super EBA was less influenced by storage medium than the other materials, especially MTA. However, further long-term studies considering other factors, such as biocompatibility (i.e. cellular toxicity) and retention, are needed to be collaborated with these findings in the clinical context.

Comparison of the microhardness and morphology of five different retrograde filling materials in aqueous and dry conditions.

The purpose of the present study was to compare the effect of dry and aqueous conditions on the surface morphology and surface hardness of five materials 24 h after being used as fillings without initial setting time in dry condition. The five materials were ProRoot mineral trioxide aggregate (MTA), super EBA, intermediate restorative materials (IRM), zinc oxide eugenol (ZOE), and amalgam. To evaluate microhardness, the five materials were submitted to the Vickers microhardness (VHN) test. We used a scanning electron microscope (Steroscan 440, Leica Cambridge, England) to observe the microstructural morphology of the five different materials. The VHN of MTA soaked in water showed five times lower than that of MTA soaked in dry condition. On the other hand, super EBA was less influenced by the medium of storage compared with the other materials. Scanning electron microscope (SEM) images showed the similar results with microhardness tests. The surface of MTA soaked in water appeared to be unstable compared with that of dry condition while super EBA showed similarly smooth surface in both conditions (aqueous and dry). In conclusion, the physical property of MTA is reduced after storage in water; however, super EBA is less influenced by aqueous condition.