Different concentrations of fetal bovine serum affect cytokine modulation in Lipopolysaccharide-activated apical papilla cells in vitro
Abstract Fetal bovine serum (FBS) is the most used supplement in culture media; however, it may interfere with in vitro assays via effects on cell proliferation and cytokine production. The ideal FBS concentration for assays using apical papilla cells (APCs) remains unknown. Therefore, this study ai...
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| Format: | Article |
| Language: | English |
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University of São Paulo
2023-07-01
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| Series: | Journal of Applied Oral Science |
| Subjects: | |
| Online Access: | http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1678-77572023000100435&lng=en&tlng=en |
| _version_ | 1797773064371437568 |
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| author | Letícia Martins SANTOS Patricia e Silva CARDOSO Elisa Abreu DINIZ Juliana Garuba RAHHAL Carla Renata SIPERT |
| author_facet | Letícia Martins SANTOS Patricia e Silva CARDOSO Elisa Abreu DINIZ Juliana Garuba RAHHAL Carla Renata SIPERT |
| author_sort | Letícia Martins SANTOS |
| collection | DOAJ |
| description | Abstract Fetal bovine serum (FBS) is the most used supplement in culture media; however, it may interfere with in vitro assays via effects on cell proliferation and cytokine production. The ideal FBS concentration for assays using apical papilla cells (APCs) remains unknown. Therefore, this study aimed to evaluate the effects of FBS on APC activation, cell viability/proliferation, and cytokine production. Methodology Human APCs were cultured, plated, and maintained in media containing increasing concentrations of FBS for 24 h, 48 h, 72 h, 7 days, and 14 days in the presence of Lipopolysaccharide (LPS - 1 µg/mL). At each time point, the cells were subjected to the MTT assay. The cytokines transforming growth factor (TGF)-β1, osteoprotegerin (OPG), and interleukin (IL)-6, along with the chemokine CCL2, were quantified using the enzyme-linked immunosorbent assay at the 24-h time-point. Statistical analysis was performed using two-way analysis of variance (ANOVA) followed by Tukey's post-hoc test (p<0.05). Results In general, APCs exhibited increasing metabolic activity in an FBS concentration-dependent fashion, regardless of the presence of LPS. In contrast, FBS interfered with the production of all the cytokines evaluated in this study, affecting the response induced by the presence of LPS. Conclusion FBS increased APC metabolism in a concentration-dependent manner and differentially affected the production of TGF-β1, OPG, IL-6, and CCL2 by APCs in vitro. |
| first_indexed | 2024-03-12T22:00:51Z |
| format | Article |
| id | doaj.art-348c07bbadc645ebbc91a6f135258978 |
| institution | Directory Open Access Journal |
| issn | 1678-7765 |
| language | English |
| last_indexed | 2024-03-12T22:00:51Z |
| publishDate | 2023-07-01 |
| publisher | University of São Paulo |
| record_format | Article |
| series | Journal of Applied Oral Science |
| spelling | doaj.art-348c07bbadc645ebbc91a6f1352589782023-07-25T07:46:15ZengUniversity of São PauloJournal of Applied Oral Science1678-77652023-07-013110.1590/1678-7757-2023-0020Different concentrations of fetal bovine serum affect cytokine modulation in Lipopolysaccharide-activated apical papilla cells in vitroLetícia Martins SANTOShttps://orcid.org/0000-0002-7492-3671Patricia e Silva CARDOSOhttps://orcid.org/0000-0002-6339-4057Elisa Abreu DINIZJuliana Garuba RAHHALhttps://orcid.org/0000-0003-2761-8188Carla Renata SIPERThttps://orcid.org/0000-0002-5719-6505Abstract Fetal bovine serum (FBS) is the most used supplement in culture media; however, it may interfere with in vitro assays via effects on cell proliferation and cytokine production. The ideal FBS concentration for assays using apical papilla cells (APCs) remains unknown. Therefore, this study aimed to evaluate the effects of FBS on APC activation, cell viability/proliferation, and cytokine production. Methodology Human APCs were cultured, plated, and maintained in media containing increasing concentrations of FBS for 24 h, 48 h, 72 h, 7 days, and 14 days in the presence of Lipopolysaccharide (LPS - 1 µg/mL). At each time point, the cells were subjected to the MTT assay. The cytokines transforming growth factor (TGF)-β1, osteoprotegerin (OPG), and interleukin (IL)-6, along with the chemokine CCL2, were quantified using the enzyme-linked immunosorbent assay at the 24-h time-point. Statistical analysis was performed using two-way analysis of variance (ANOVA) followed by Tukey's post-hoc test (p<0.05). Results In general, APCs exhibited increasing metabolic activity in an FBS concentration-dependent fashion, regardless of the presence of LPS. In contrast, FBS interfered with the production of all the cytokines evaluated in this study, affecting the response induced by the presence of LPS. Conclusion FBS increased APC metabolism in a concentration-dependent manner and differentially affected the production of TGF-β1, OPG, IL-6, and CCL2 by APCs in vitro.http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1678-77572023000100435&lng=en&tlng=enCytokinesFetal bovine serumApical papilla cells |
| spellingShingle | Letícia Martins SANTOS Patricia e Silva CARDOSO Elisa Abreu DINIZ Juliana Garuba RAHHAL Carla Renata SIPERT Different concentrations of fetal bovine serum affect cytokine modulation in Lipopolysaccharide-activated apical papilla cells in vitro Journal of Applied Oral Science Cytokines Fetal bovine serum Apical papilla cells |
| title | Different concentrations of fetal bovine serum affect cytokine modulation in Lipopolysaccharide-activated apical papilla cells in vitro |
| title_full | Different concentrations of fetal bovine serum affect cytokine modulation in Lipopolysaccharide-activated apical papilla cells in vitro |
| title_fullStr | Different concentrations of fetal bovine serum affect cytokine modulation in Lipopolysaccharide-activated apical papilla cells in vitro |
| title_full_unstemmed | Different concentrations of fetal bovine serum affect cytokine modulation in Lipopolysaccharide-activated apical papilla cells in vitro |
| title_short | Different concentrations of fetal bovine serum affect cytokine modulation in Lipopolysaccharide-activated apical papilla cells in vitro |
| title_sort | different concentrations of fetal bovine serum affect cytokine modulation in lipopolysaccharide activated apical papilla cells in vitro |
| topic | Cytokines Fetal bovine serum Apical papilla cells |
| url | http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1678-77572023000100435&lng=en&tlng=en |
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