CCL3 and CXCL12 production in vitro by dental pulp fibroblasts from permanent and deciduous teeth stimulated by Porphyromonas gingivalis LPS

ABSTRACT Objective: The aim of this study was to compare the production of the chemokines CCL3 and CXCL12 by cultured dental pulp fibroblasts from permanent (PDPF) and deciduous (DDPF) teeth under stimulation by Porphyromonas gingivalis LPS (PgLPS). Material and Methods: Primary culture of fibroblasts from permanent (n=3) and deciduous (n=2) teeth were established using an explant technique. After the fourth passage, fibroblasts were stimulated by increasing concentrations of PgLPS (0 – 10 µg/mL) at 1, 6 and 24 h. The cells were tested for viability through MTT assay, and production of the chemokines CCL3 and CXCL12 was determined through ELISA. Comparisons among samples were performed using One-way ANOVA for MTT assay and Two-way ANOVA for ELISA results. Results: Cell viability was not affected by the antigen after 24 h of stimulation. PgLPS induced the production of CCL3 by dental pulp fibroblasts at similar levels for both permanent and deciduous pulp fibroblasts. Production of CXCL12, however, was significantly higher for PDPF than DDPF at 1 and 6 h. PgLPS, in turn, downregulated the production of CXCL12 by PDPF but not by DDPF. Conclusion: These data suggest that dental pulp fibroblasts from permanent and deciduous teeth may present a differential behavior under PgLPS stimulation. .

Immunohistochemical assessment of lymphangiogenesis in inflamed human dental pulp

The present study aimed to investigate the effect of inflammation on lymphangiogenesis in human dental pulp using monoclonal mouse anti-human D2-40 antibody. Thirty samples of healthy dental pulps and twenty-seven pulps with irreversible pulpitis were included in this study. All cases were immunohistochemically stained for D2-40. Positively stained lymphatic vessels were counted by a light microscopy in an area of 1 mm2. The mean number and standard deviation of lymphatic vessels positive for D2-40 in the group with inflammation (8.04 ± 2.8) was significantly higher (p< 0.0001) than the mean number in the group without inflammation (3.93 ± 1.11). The study established an increased number of lymphatic vessels in the inflamed human dental pulp, which suggests that inflammation contributes lymphangiogenesis. The new lymphatic vessels can accelerate the removal of interstitial fluid and thus limit the effects of increased tissue pressure on the connective tissue. This may have importance in the process of regeneration of dental pulp and conservative procedures such as pulpotomy.