Matrix metalloproteinase response of dendritic cell, gingival epithelial keratinocyte, and T-cell transwell co-cultures treated with porphyromonas gingivalis hemagglutinin-B

Amber M. Bates, Carol L. Fischer, Vrushali Abhyankar, Georgia K. Johnson, Janet M. Guthmiller, Ann Progulske-Fox, Kim A. Brogden

Research output: Contribution to journalArticle

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Abstract

Matrix metalloproteinases (MMPs) are enzymes involved in periodontal tissue destruction. Hemagglutinin B (HagB) from the periodontal pathogen Porphyromonas gingivalis induces an elevated MMP response in dendritic cells, but responses from cultures of single-cell types do not reflect the local tissue environment. The objective of this study was to measure HagB-induced MMP responses in a transwell co-culture system containing dendritic cells, gingival epithelial (GE) keratinocytes, and CD4+ T-cells. Transwell co-cultures were assembled and treated with or without HagB. Immunoassays were used to determine production of MMP1, MMP7, MMP9, and MMP12 in response to HagB up to 64 h. Control responses were subtracted from HagB-induced responses. A two-way fixed effect ANOVA was fit to log-transformed concentrations and pairwise group comparisons were conducted (p < 0.05). At 64 h, dendritic cells produced elevated MMP1 and MMP9 responses, which were attenuated in the 3-cell co-culture (p < 0.05). There were also significant differences in MMP7 and MMP12 production between single-cell cultures and co-cultures. These results support the need to use multiple cell types in culture models to evaluate a more representative response to proinflammatory agonists. This three-cell transwell co-culture model may help us better understand the inflammatory process in periodontal disease and test novel therapeutic approaches.

Original languageEnglish (US)
Article number3923
JournalInternational journal of molecular sciences
Volume19
Issue number12
DOIs
StatePublished - Dec 1 2018

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T-cells
Hemagglutinins
Coculture Techniques
Matrix Metalloproteinases
Keratinocytes
Cell culture
Dendritic Cells
T-Lymphocytes
Cell Culture Techniques
matrices
cells
Tissue
Pathogens
Analysis of variance (ANOVA)
Enzymes
Porphyromonas gingivalis
Periodontal Diseases
Immunoassay
Analysis of Variance
immunoassay

All Science Journal Classification (ASJC) codes

  • Catalysis
  • Molecular Biology
  • Spectroscopy
  • Computer Science Applications
  • Physical and Theoretical Chemistry
  • Organic Chemistry
  • Inorganic Chemistry

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Matrix metalloproteinase response of dendritic cell, gingival epithelial keratinocyte, and T-cell transwell co-cultures treated with porphyromonas gingivalis hemagglutinin-B. / Bates, Amber M.; Fischer, Carol L.; Abhyankar, Vrushali; Johnson, Georgia K.; Guthmiller, Janet M.; Progulske-Fox, Ann; Brogden, Kim A.

In: International journal of molecular sciences, Vol. 19, No. 12, 3923, 01.12.2018.

Research output: Contribution to journalArticle

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abstract = "Matrix metalloproteinases (MMPs) are enzymes involved in periodontal tissue destruction. Hemagglutinin B (HagB) from the periodontal pathogen Porphyromonas gingivalis induces an elevated MMP response in dendritic cells, but responses from cultures of single-cell types do not reflect the local tissue environment. The objective of this study was to measure HagB-induced MMP responses in a transwell co-culture system containing dendritic cells, gingival epithelial (GE) keratinocytes, and CD4+ T-cells. Transwell co-cultures were assembled and treated with or without HagB. Immunoassays were used to determine production of MMP1, MMP7, MMP9, and MMP12 in response to HagB up to 64 h. Control responses were subtracted from HagB-induced responses. A two-way fixed effect ANOVA was fit to log-transformed concentrations and pairwise group comparisons were conducted (p < 0.05). At 64 h, dendritic cells produced elevated MMP1 and MMP9 responses, which were attenuated in the 3-cell co-culture (p < 0.05). There were also significant differences in MMP7 and MMP12 production between single-cell cultures and co-cultures. These results support the need to use multiple cell types in culture models to evaluate a more representative response to proinflammatory agonists. This three-cell transwell co-culture model may help us better understand the inflammatory process in periodontal disease and test novel therapeutic approaches.",
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