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Increased Eotaxin and MCP-1 Levels in Serum from Individuals with Periodontitis and in Human Gingival Fibroblasts Exposed to Pro-Inflammatory Cytokines.

Boström EA, Kindstedt E, Sulniute R, Palmqvist P, Majster M, Holm CK, Zwicker S, Clark R, Önell S, Johansson I, Lerner UH, Lundberg P - PLoS ONE (2015)

Bottom Line: Moreover, body mass index correlated significantly with serum MCP-1 and CRP, but not with eotaxin.We detected higher MCP-1 protein levels in inflamed gingival connective tissue compared to healthy but the eotaxin levels were undetectable.Primary human gingival fibroblasts displayed strongly increased expression of MCP-1 and eotaxin mRNA and protein when challenged with tumor necrosis factor-α (TNF-α and interleukin-1β (IL-1β), key mediators of periodontal inflammation.

View Article: PubMed Central - PubMed

Affiliation: Karolinska Institutet, Division of Periodontology, Department of Dental Medicine, Stockholm, Sweden.

ABSTRACT
Periodontitis is a chronic inflammatory disease of tooth supporting tissues resulting in periodontal tissue destruction, which may ultimately lead to tooth loss. The disease is characterized by continuous leukocyte infiltration, likely mediated by local chemokine production but the pathogenic mechanisms are not fully elucidated. There are no reliable serologic biomarkers for the diagnosis of periodontitis, which is today based solely on the degree of local tissue destruction, and there is no available biological treatment tool. Prompted by the increasing interest in periodontitis and systemic inflammatory mediators we mapped serum cytokine and chemokine levels from periodontitis subjects and healthy controls. We used multivariate partial least squares (PLS) modeling and identified monocyte chemoattractant protein-1 (MCP-1) and eotaxin as clearly associated with periodontitis along with C-reactive protein (CRP), years of smoking and age, whereas the number of remaining teeth was associated with being healthy. Moreover, body mass index correlated significantly with serum MCP-1 and CRP, but not with eotaxin. We detected higher MCP-1 protein levels in inflamed gingival connective tissue compared to healthy but the eotaxin levels were undetectable. Primary human gingival fibroblasts displayed strongly increased expression of MCP-1 and eotaxin mRNA and protein when challenged with tumor necrosis factor-α (TNF-α and interleukin-1β (IL-1β), key mediators of periodontal inflammation. We also demonstrated that the upregulated chemokine expression was dependent on the NF-κΒ pathway. In summary, we identify higher levels of CRP, eotaxin and MCP-1 in serum of periodontitis patients. This, together with our finding that both CRP and MCP-1 correlates with BMI points towards an increased systemic inflammatory load in patients with periodontitis and high BMI. Targeting eotaxin and MCP-1 in periodontitis may result in reduced leukocyte infiltration and inflammation in periodontitis and maybe prevent tooth loss.

No MeSH data available.


Related in: MedlinePlus

Time- and dose-dependent increase of eotaxin and MCP-1 protein expression in gingival fibroblasts stimulated by TNF-α and IL-1β.(A and C) TNF-α (50 ng/ml) stimulates eotaxin and MCP-1 protein expression, and (B and D) IL-1β (100 pg/ml) stimulates eotaxin and MCP-1 protein expression a time-dependent manner. (E and G) TNF-α stimulates eotaxin and MCP-1 protein expression, and (F and H) IL-1β stimulates eotaxin and MCP-1 protein expression in a dose-dependent manner.
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pone.0134608.g003: Time- and dose-dependent increase of eotaxin and MCP-1 protein expression in gingival fibroblasts stimulated by TNF-α and IL-1β.(A and C) TNF-α (50 ng/ml) stimulates eotaxin and MCP-1 protein expression, and (B and D) IL-1β (100 pg/ml) stimulates eotaxin and MCP-1 protein expression a time-dependent manner. (E and G) TNF-α stimulates eotaxin and MCP-1 protein expression, and (F and H) IL-1β stimulates eotaxin and MCP-1 protein expression in a dose-dependent manner.

Mentions: Given that gingival fibroblasts are the most abundant cell in gingival connective tissue they are potential contributors to chemokine production in periodontitis. We therefore assessed if isolated gingival fibroblasts constitutively expressed eotaxin and MCP-1 and if their expression was regulated in response to pro-inflammatory stimuli. Human gingival fibroblasts were cultured at increasing time-points (1h, 3h, 6h, 24h, 48h and 72h) in the absence or presence of TNF-α (50 ng/ml) or IL-1β (100 pg/ml). Protein analysis using ELISA showed that the cells constitutively expressed eotaxin and MCP-1 protein. Treatment of the cells with IL-1β or TNF-α resulted in a time-dependent upregulation of eotaxin expression with statistically significant increase at 6h in the presence of either TNF-α (P<0.05) (Fig 3A), or IL-1β (P<0.01) (Fig 3B). A time-dependent upregulation of MCP-1 protein expression was also seen in cells incubated with TNF-α (Fig 3C) or IL-1β (Fig 3D) with a significant (P<0.001) effect observed after 3h with TNF-α or IL-1β exposure.


Increased Eotaxin and MCP-1 Levels in Serum from Individuals with Periodontitis and in Human Gingival Fibroblasts Exposed to Pro-Inflammatory Cytokines.

Boström EA, Kindstedt E, Sulniute R, Palmqvist P, Majster M, Holm CK, Zwicker S, Clark R, Önell S, Johansson I, Lerner UH, Lundberg P - PLoS ONE (2015)

Time- and dose-dependent increase of eotaxin and MCP-1 protein expression in gingival fibroblasts stimulated by TNF-α and IL-1β.(A and C) TNF-α (50 ng/ml) stimulates eotaxin and MCP-1 protein expression, and (B and D) IL-1β (100 pg/ml) stimulates eotaxin and MCP-1 protein expression a time-dependent manner. (E and G) TNF-α stimulates eotaxin and MCP-1 protein expression, and (F and H) IL-1β stimulates eotaxin and MCP-1 protein expression in a dose-dependent manner.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4524692&req=5

pone.0134608.g003: Time- and dose-dependent increase of eotaxin and MCP-1 protein expression in gingival fibroblasts stimulated by TNF-α and IL-1β.(A and C) TNF-α (50 ng/ml) stimulates eotaxin and MCP-1 protein expression, and (B and D) IL-1β (100 pg/ml) stimulates eotaxin and MCP-1 protein expression a time-dependent manner. (E and G) TNF-α stimulates eotaxin and MCP-1 protein expression, and (F and H) IL-1β stimulates eotaxin and MCP-1 protein expression in a dose-dependent manner.
Mentions: Given that gingival fibroblasts are the most abundant cell in gingival connective tissue they are potential contributors to chemokine production in periodontitis. We therefore assessed if isolated gingival fibroblasts constitutively expressed eotaxin and MCP-1 and if their expression was regulated in response to pro-inflammatory stimuli. Human gingival fibroblasts were cultured at increasing time-points (1h, 3h, 6h, 24h, 48h and 72h) in the absence or presence of TNF-α (50 ng/ml) or IL-1β (100 pg/ml). Protein analysis using ELISA showed that the cells constitutively expressed eotaxin and MCP-1 protein. Treatment of the cells with IL-1β or TNF-α resulted in a time-dependent upregulation of eotaxin expression with statistically significant increase at 6h in the presence of either TNF-α (P<0.05) (Fig 3A), or IL-1β (P<0.01) (Fig 3B). A time-dependent upregulation of MCP-1 protein expression was also seen in cells incubated with TNF-α (Fig 3C) or IL-1β (Fig 3D) with a significant (P<0.001) effect observed after 3h with TNF-α or IL-1β exposure.

Bottom Line: Moreover, body mass index correlated significantly with serum MCP-1 and CRP, but not with eotaxin.We detected higher MCP-1 protein levels in inflamed gingival connective tissue compared to healthy but the eotaxin levels were undetectable.Primary human gingival fibroblasts displayed strongly increased expression of MCP-1 and eotaxin mRNA and protein when challenged with tumor necrosis factor-α (TNF-α and interleukin-1β (IL-1β), key mediators of periodontal inflammation.

View Article: PubMed Central - PubMed

Affiliation: Karolinska Institutet, Division of Periodontology, Department of Dental Medicine, Stockholm, Sweden.

ABSTRACT
Periodontitis is a chronic inflammatory disease of tooth supporting tissues resulting in periodontal tissue destruction, which may ultimately lead to tooth loss. The disease is characterized by continuous leukocyte infiltration, likely mediated by local chemokine production but the pathogenic mechanisms are not fully elucidated. There are no reliable serologic biomarkers for the diagnosis of periodontitis, which is today based solely on the degree of local tissue destruction, and there is no available biological treatment tool. Prompted by the increasing interest in periodontitis and systemic inflammatory mediators we mapped serum cytokine and chemokine levels from periodontitis subjects and healthy controls. We used multivariate partial least squares (PLS) modeling and identified monocyte chemoattractant protein-1 (MCP-1) and eotaxin as clearly associated with periodontitis along with C-reactive protein (CRP), years of smoking and age, whereas the number of remaining teeth was associated with being healthy. Moreover, body mass index correlated significantly with serum MCP-1 and CRP, but not with eotaxin. We detected higher MCP-1 protein levels in inflamed gingival connective tissue compared to healthy but the eotaxin levels were undetectable. Primary human gingival fibroblasts displayed strongly increased expression of MCP-1 and eotaxin mRNA and protein when challenged with tumor necrosis factor-α (TNF-α and interleukin-1β (IL-1β), key mediators of periodontal inflammation. We also demonstrated that the upregulated chemokine expression was dependent on the NF-κΒ pathway. In summary, we identify higher levels of CRP, eotaxin and MCP-1 in serum of periodontitis patients. This, together with our finding that both CRP and MCP-1 correlates with BMI points towards an increased systemic inflammatory load in patients with periodontitis and high BMI. Targeting eotaxin and MCP-1 in periodontitis may result in reduced leukocyte infiltration and inflammation in periodontitis and maybe prevent tooth loss.

No MeSH data available.


Related in: MedlinePlus