Limits...
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.

Show MeSH

Related in: MedlinePlus

Cell survival under anaerobic conditions.Survival of hDFSC, hBMSC, hGiF and Ca9-22 cells under anaerobic compared with aerobic conditions. Anaerobic atmosphere: (10% CO2, 10% H2, 80% N2) and 37°C. Viable cells were counted at timepoints of 24, 48 and 72 h in a Neubauer hemacytometer by exclusion of trypan blue. Values are expressed as means ± SD (standard deviation), *p<0.05 (T-test), n = 4. Asterisks indicate statistically significant differences between hDFSC and hBMSC, hGiF, and Ca9-22.
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4219685&req=5

pone-0110616-g001: Cell survival under anaerobic conditions.Survival of hDFSC, hBMSC, hGiF and Ca9-22 cells under anaerobic compared with aerobic conditions. Anaerobic atmosphere: (10% CO2, 10% H2, 80% N2) and 37°C. Viable cells were counted at timepoints of 24, 48 and 72 h in a Neubauer hemacytometer by exclusion of trypan blue. Values are expressed as means ± SD (standard deviation), *p<0.05 (T-test), n = 4. Asterisks indicate statistically significant differences between hDFSC and hBMSC, hGiF, and Ca9-22.

Mentions: The establishment of the anaerobic co-culture system as described by Kriebel et al. [21] included the verification of the cell survival under an anaerobic atmosphere prior to a bacterial challenge. Survival of the cells was evaluated after 12, 24 and 72 h (Fig. 1). Compared to aerobic conditions, 81.7% of the hDFSC, 83.2% of the hBMSC, 54.0% of the Ca9-22 cells, and 58.9% of the hGiF remained viable 24 h of an oxygen-free incubation. After 2 days, the hDFSC still showed higher survival rates than the Ca9-22: only 33.8% of Ca9-22 cells survived the 48-h incubation, whereas 50.9% of hDFSC cells were alive. The cell numbers decreased significantly for all cells after 72 h: hDFSC showed a survival of 40.2%, hBMSC 35.0%, hGiF 16.1%, and Ca9-22 13.5% (Fig. 1).


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

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

Cell survival under anaerobic conditions.Survival of hDFSC, hBMSC, hGiF and Ca9-22 cells under anaerobic compared with aerobic conditions. Anaerobic atmosphere: (10% CO2, 10% H2, 80% N2) and 37°C. Viable cells were counted at timepoints of 24, 48 and 72 h in a Neubauer hemacytometer by exclusion of trypan blue. Values are expressed as means ± SD (standard deviation), *p<0.05 (T-test), n = 4. Asterisks indicate statistically significant differences between hDFSC and hBMSC, hGiF, and Ca9-22.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4219685&req=5

pone-0110616-g001: Cell survival under anaerobic conditions.Survival of hDFSC, hBMSC, hGiF and Ca9-22 cells under anaerobic compared with aerobic conditions. Anaerobic atmosphere: (10% CO2, 10% H2, 80% N2) and 37°C. Viable cells were counted at timepoints of 24, 48 and 72 h in a Neubauer hemacytometer by exclusion of trypan blue. Values are expressed as means ± SD (standard deviation), *p<0.05 (T-test), n = 4. Asterisks indicate statistically significant differences between hDFSC and hBMSC, hGiF, and Ca9-22.
Mentions: The establishment of the anaerobic co-culture system as described by Kriebel et al. [21] included the verification of the cell survival under an anaerobic atmosphere prior to a bacterial challenge. Survival of the cells was evaluated after 12, 24 and 72 h (Fig. 1). Compared to aerobic conditions, 81.7% of the hDFSC, 83.2% of the hBMSC, 54.0% of the Ca9-22 cells, and 58.9% of the hGiF remained viable 24 h of an oxygen-free incubation. After 2 days, the hDFSC still showed higher survival rates than the Ca9-22: only 33.8% of Ca9-22 cells survived the 48-h incubation, whereas 50.9% of hDFSC cells were alive. The cell numbers decreased significantly for all cells after 72 h: hDFSC showed a survival of 40.2%, hBMSC 35.0%, hGiF 16.1%, and Ca9-22 13.5% (Fig. 1).

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