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. 2013 Mar-Apr;21(2):99-105.
doi: 10.1590/1678-7757201300004.

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

Carla Renata Sipert  1 , Ana Carolina de Faria Morandini, Karin Cristina da Silva Modena, Thiago José Dionísio, Maria Aparecida Andrade Moreira Machado, Sandra Helena Penha de Oliveira, Ana Paula Campanelli, Carlos Ferreira Santos
Affiliations

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

Carla Renata Sipert et al. J Appl Oral Sci. 2013 Mar-Apr.

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.

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Figures

Figure 1
Figure 1
Characterization of dental pulp fibroblasts by procollagen I and fibroblast surface protein staining. Cultured dental pulp fibroblasts from permanent (n=1) and deciduous (n=1) teeth were immunostained for procollagen I (A and B) or fibroblast surface protein (C and D). A and C: PDPF; B and D: DDPF. Images captured by a confocal microscope (representative bars: 20 μm).
Figure 2
Figure 2
Cell viability of dental pulp fibroblasts challenged with P. gingivalis LPS. Cultured dental pulp fibroblasts from permanent (n=1; panel A) and deciduous (n=1, panel B) teeth were stimulated by P. gingivalis LPS (PgLPS) at the indicated concentrations in triplicate. Cell viability was assessed by means of MTT assay (n=3).
Figure 3
Figure 3
Production of chemokines by different subtypes of dental pulp fibroblasts. Cultured dental pulp fibroblasts from permanent (n=3) and deciduous (n=2) teeth were stimulated by P. gingivalis LPS at the indicated concentrations (n=6). Cell supernatants were collected after 1 (A and D), 6 (B and E) and 24 (C and F) h. Production of CCL3 (A, B and C) and CXCL12 (D, E and F) was detected by means of ELISA. (*) p<0.05; (**) p<0.01 and (***) p<0.001 in comparison with culture medium alone (0). (##) p<0.01 and (###) p<0.001 in comparison with the other cellular subtype at the same experimental condition.
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Comment in

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