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Hyaluronan-mediated mononuclear leukocyte binding to gingival fibroblasts

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Abstract

Objectives

Binding of mononuclear leukocytes to hyaluronan cable structures is a well-known pathomechanism in several chronic inflammatory diseases, but has not yet described for chronic oral inflammations. The aim of this study was to evaluate if and how binding of mononuclear leukocytes to pathologic hyaluronan cable structures can be induced in human gingival fibroblasts.

Material and methods

Experiments were performed with human gingival fibroblasts and peripheral blood mononuclear cells (PBMCs) from three healthy blood donors. Gingival fibroblasts were stimulated with (1) tunicamycin, (2) polyinosinic/polycytidylic acid (Poly:IC), and (3) lipopolysaccharides (LPS) to simulate (1) ER stress and (2) viral and (3) bacterial infections, respectively. Fibroblasts were then co-incubated with PBMCs, and the number of bound and fluorescently labeled PBMCs was assessed using a fluorescence reader and microscopy. For data analysis, a linear mixed model was used.

Results

Hyaluronan-mediated binding of PBMCs to gingival fibroblasts was increased by tunicamycin and Poly(I:C) but not by LPS. Hyaluronidase treatment and co-incubation with hyaluronan transport inhibitors reduced this binding.

Conclusions

Results suggest that hyaluronan-mediated binding of blood cells might play a role in oral inflammations. A potential superior role of viruses needs to be confirmed in further clinical studies.

Clinical relevance

The linkage between pathological hyaluronan matrices and oral infections opens up potential applications of hyaluronan transport inhibitors in the treatment of chronic oral inflammations.

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Acknowledgements

The authors thank Aline Saehr and Sandra Foermer for excellent technical assistance and Tess Blundell for English language editing. Special thanks go to Klaus Heeg for providing infrastructure for cell culture experiments in Heidelberg.

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Authors and Affiliations

  1. Section of Periodontology, Department of Conservative Dentistry, Clinic for Oral, Dental and Maxillofacial Diseases, Heidelberg University Hospital, Im Neuenheimer Feld 400, 69120, Heidelberg, Germany

    Daniel Hagenfeld & Ti-Sun Kim

  2. Department of Periodontology and Conservative Dentistry, Muenster University Hospital, Albert Schweitzer Campus 1, Building W30, 48149, Münster, Germany

    Daniel Hagenfeld & Inga Harks

  3. Department of Infectious Diseases, Heidelberg University Hospital, Im Neuenheimer Feld 324, 69120, Heidelberg, Germany

    Nico T. Mutters

  4. Institute of Biostatistics and Clinical Research, University of Muenster, Schmeddingstraße 56, 48149, Münster, Germany

    Raphael Koch

  5. Hylitis, Rudolf-Harbig-Straße 5, 48301, Nottuln, Germany

    Peter Prehm

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  1. Daniel Hagenfeld
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Contributions

Conceived and designed this study: DH and PP. Performed experiments: DH. Conceived and designed analysis: DH. Statistical analysis: DH and RK. First draft of manuscript: DH and NM. Contributed and approved the final version of the manuscript: DH, NM, IH, RK, TSK, and PP.

Corresponding author

Correspondence to Daniel Hagenfeld.

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Conflict of interest

The authors declare that they have no conflicts of interest.

Funding

The Medical Faculty of Heidelberg supported this work with a postdoc fellowship for DH.

Ethical approval

All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional research committee and with the 1964 Helsinki Declaration and its later amendments or comparable ethical standards.

Informed consent

Informed consent was obtained from all individual participants included in the study.

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Suppl. Fig. 1

Hyaluronan distribution in inflamed and non-inflamed gingival tissue Upper part: Confocal laser scanning microscopy of connective tissue of a patient after conservative periodontal treatment (a) and a patient without periodontal treatment (b). Nuclei stained with HOECHST 33342 (blue). Auto-fluorescence of gingival connective tissue (green). Hyaluronan stained with biotinylated hyaluronan binding protein (red). Lower part: Staining of fibroblast-PBMC co-culture after tunicamycin treatment for 10 minutes (c) and incubation after tunicamycin treatment with 5U hyaluronidase for 10min (d) nuclei (blue), hyaluronan binding protein (green) and CD44 surface marker (red). (JPEG 98 kb)

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Hagenfeld, D., Mutters, N.T., Harks, I. et al. Hyaluronan-mediated mononuclear leukocyte binding to gingival fibroblasts. Clin Oral Invest 22, 1063–1070 (2018). https://doi.org/10.1007/s00784-017-2188-x

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  • DOI: https://doi.org/10.1007/s00784-017-2188-x

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