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Interactions of anaerobic bacteria with dental stem cells: an in vitro study.

Biedermann A, Kriebel K, Kreikemeyer B, Lang H - PLoS ONE (2014)

Bottom Line: Similar results were observed for host cell internalization (F. nucleatum: 0.002% internalization into hDFSC; 0.09% internalization into Ca9-22).The IL-10 response of the differentiated cells was found to be low in relation to their pro-inflammatory IL-8 response.The results indicate that dental stem cells are less prone to interactions with pathogenic bacteria than differentiated cells in an anaerobic environment.

View Article: PubMed Central - PubMed

Affiliation: Department of Operative Dentistry and Periodontology, University of Rostock, Rostock, Germany.

ABSTRACT

Background: In patients with periodontitis, it is highly likely that local (progenitor) cells encounter pathogenic bacteria. The purpose of this in vitro study was to elucidate how human dental follicle stem cells (hDFSC) react towards a direct challenge with anaerobic periodontal pathogens under their natural oxygen-free atmosphere. HDFSC were compared to human bone marrow mesenchymal stem cells (hBMSC) and differentiated primary human gingival fibroblasts (hGiF), as well as permanent gingival carcinoma cells (Ca9-22).

Methodology/principal findings: The different cell types were investigated in a co-culture system with Porphyromonas gingivalis (P. gingivalis) and Fusobacterium nucleatum (F. nucleatum). The viability of the cells and pathogens under anaerobic conditions, as well as interactions in terms of adherence and internalization, were examined. Additionally, the release of pro-inflammatory interleukin-8 (IL-8) and anti-inflammatory interleukin-10 (IL-10) was quantified via enzyme-linked immunosorbent assay. The bacteria adhered less efficiently to hDFSC compared to Ca9-22 (P. gingivalis: 0.18% adherence to hDFSC; 3.1% adherence to Ca9-22). Similar results were observed for host cell internalization (F. nucleatum: 0.002% internalization into hDFSC; 0.09% internalization into Ca9-22). Statistically significantly less IL-8 was secreted from hDFSC after stimulation with F. nucleatum and P. gingivalis in comparison with hGiF (F. nucleatum: 2080.0 pg/ml--hGiF; 19.7 pg/ml--hDFSC). The IL-10 response of the differentiated cells was found to be low in relation to their pro-inflammatory IL-8 response.

Conclusions/significance: The results indicate that dental stem cells are less prone to interactions with pathogenic bacteria than differentiated cells in an anaerobic environment. Moreover, during bacterial challenge, the stem cell immune response seems to be more towards an anti-inflammatory reaction. For a potential future therapeutic use of hDFSC, these findings support the idea of a save application.

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IL-8 and IL-10 secretion.The assays were carried out as described in the text. IL-8 and IL-10 levels were assayed by ELISA. Absorbance was read at 450 nm. Values represent the means ± SD, *p<0.05, **p = 0.01 (T-test), n = 4. Asterisks indicate statistically significant differences between hDFSC and hBMSC, hGiF, and Ca9-22. Cytokine secretion by hDFSC was compared with secretion by hBMSC, hGiF, and Ca9-22 cells at the same timepoint. A) IL-8 measured in the supernatant of the F.nucleatum ATCC 23727 and 25586 co-culture after 1, 2, 4, and 24 h. B) IL-8 measured in the supernatant of the P. gingivalis W50 and W83 co-culture after 1, 2, 4, and 24 h. C) IL-10 measured in the supernatant of the F.nucleatum ATCC 23727 and 25586 co-culture after 1, 2, 4, and 24 h. D) IL-10 measured in the supernatant of the P. gingivalis W50 and W83 co-culture after 1, 2, 4, and 24 h.
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pone-0110616-g004: IL-8 and IL-10 secretion.The assays were carried out as described in the text. IL-8 and IL-10 levels were assayed by ELISA. Absorbance was read at 450 nm. Values represent the means ± SD, *p<0.05, **p = 0.01 (T-test), n = 4. Asterisks indicate statistically significant differences between hDFSC and hBMSC, hGiF, and Ca9-22. Cytokine secretion by hDFSC was compared with secretion by hBMSC, hGiF, and Ca9-22 cells at the same timepoint. A) IL-8 measured in the supernatant of the F.nucleatum ATCC 23727 and 25586 co-culture after 1, 2, 4, and 24 h. B) IL-8 measured in the supernatant of the P. gingivalis W50 and W83 co-culture after 1, 2, 4, and 24 h. C) IL-10 measured in the supernatant of the F.nucleatum ATCC 23727 and 25586 co-culture after 1, 2, 4, and 24 h. D) IL-10 measured in the supernatant of the P. gingivalis W50 and W83 co-culture after 1, 2, 4, and 24 h.

Mentions: After co-culture with either live F. nucleatum or P. gingivalis (MOI 1∶100) for 1, 2, 4, and 24 h, IL-8 and IL-10 was detected in the supernatant of all four cell lines by ELISA (Fig. 4 and Tables S2 and S3). Unchallenged cells were used as a negative control.


Interactions of anaerobic bacteria with dental stem cells: an in vitro study.

Biedermann A, Kriebel K, Kreikemeyer B, Lang H - PLoS ONE (2014)

IL-8 and IL-10 secretion.The assays were carried out as described in the text. IL-8 and IL-10 levels were assayed by ELISA. Absorbance was read at 450 nm. Values represent the means ± SD, *p<0.05, **p = 0.01 (T-test), n = 4. Asterisks indicate statistically significant differences between hDFSC and hBMSC, hGiF, and Ca9-22. Cytokine secretion by hDFSC was compared with secretion by hBMSC, hGiF, and Ca9-22 cells at the same timepoint. A) IL-8 measured in the supernatant of the F.nucleatum ATCC 23727 and 25586 co-culture after 1, 2, 4, and 24 h. B) IL-8 measured in the supernatant of the P. gingivalis W50 and W83 co-culture after 1, 2, 4, and 24 h. C) IL-10 measured in the supernatant of the F.nucleatum ATCC 23727 and 25586 co-culture after 1, 2, 4, and 24 h. D) IL-10 measured in the supernatant of the P. gingivalis W50 and W83 co-culture after 1, 2, 4, and 24 h.
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pone-0110616-g004: IL-8 and IL-10 secretion.The assays were carried out as described in the text. IL-8 and IL-10 levels were assayed by ELISA. Absorbance was read at 450 nm. Values represent the means ± SD, *p<0.05, **p = 0.01 (T-test), n = 4. Asterisks indicate statistically significant differences between hDFSC and hBMSC, hGiF, and Ca9-22. Cytokine secretion by hDFSC was compared with secretion by hBMSC, hGiF, and Ca9-22 cells at the same timepoint. A) IL-8 measured in the supernatant of the F.nucleatum ATCC 23727 and 25586 co-culture after 1, 2, 4, and 24 h. B) IL-8 measured in the supernatant of the P. gingivalis W50 and W83 co-culture after 1, 2, 4, and 24 h. C) IL-10 measured in the supernatant of the F.nucleatum ATCC 23727 and 25586 co-culture after 1, 2, 4, and 24 h. D) IL-10 measured in the supernatant of the P. gingivalis W50 and W83 co-culture after 1, 2, 4, and 24 h.
Mentions: After co-culture with either live F. nucleatum or P. gingivalis (MOI 1∶100) for 1, 2, 4, and 24 h, IL-8 and IL-10 was detected in the supernatant of all four cell lines by ELISA (Fig. 4 and Tables S2 and S3). Unchallenged cells were used as a negative control.

Bottom Line: Similar results were observed for host cell internalization (F. nucleatum: 0.002% internalization into hDFSC; 0.09% internalization into Ca9-22).The IL-10 response of the differentiated cells was found to be low in relation to their pro-inflammatory IL-8 response.The results indicate that dental stem cells are less prone to interactions with pathogenic bacteria than differentiated cells in an anaerobic environment.

View Article: PubMed Central - PubMed

Affiliation: Department of Operative Dentistry and Periodontology, University of Rostock, Rostock, Germany.

ABSTRACT

Background: In patients with periodontitis, it is highly likely that local (progenitor) cells encounter pathogenic bacteria. The purpose of this in vitro study was to elucidate how human dental follicle stem cells (hDFSC) react towards a direct challenge with anaerobic periodontal pathogens under their natural oxygen-free atmosphere. HDFSC were compared to human bone marrow mesenchymal stem cells (hBMSC) and differentiated primary human gingival fibroblasts (hGiF), as well as permanent gingival carcinoma cells (Ca9-22).

Methodology/principal findings: The different cell types were investigated in a co-culture system with Porphyromonas gingivalis (P. gingivalis) and Fusobacterium nucleatum (F. nucleatum). The viability of the cells and pathogens under anaerobic conditions, as well as interactions in terms of adherence and internalization, were examined. Additionally, the release of pro-inflammatory interleukin-8 (IL-8) and anti-inflammatory interleukin-10 (IL-10) was quantified via enzyme-linked immunosorbent assay. The bacteria adhered less efficiently to hDFSC compared to Ca9-22 (P. gingivalis: 0.18% adherence to hDFSC; 3.1% adherence to Ca9-22). Similar results were observed for host cell internalization (F. nucleatum: 0.002% internalization into hDFSC; 0.09% internalization into Ca9-22). Statistically significantly less IL-8 was secreted from hDFSC after stimulation with F. nucleatum and P. gingivalis in comparison with hGiF (F. nucleatum: 2080.0 pg/ml--hGiF; 19.7 pg/ml--hDFSC). The IL-10 response of the differentiated cells was found to be low in relation to their pro-inflammatory IL-8 response.

Conclusions/significance: The results indicate that dental stem cells are less prone to interactions with pathogenic bacteria than differentiated cells in an anaerobic environment. Moreover, during bacterial challenge, the stem cell immune response seems to be more towards an anti-inflammatory reaction. For a potential future therapeutic use of hDFSC, these findings support the idea of a save application.

Show MeSH
Related in: MedlinePlus