Antimicrobial and anti-inflammatory activity of Cystatin C on human gingival fibroblast incubated with Porphyromonas gingivalis

PeerJ. 2022 Oct 25:10:e14232. doi: 10.7717/peerj.14232. eCollection 2022.

Abstract

Background: Periodontal disease is considered one of the most prevalent chronic infectious diseases, often leading to the disruption of tooth-supporting tissues, including alveolar bone, causing tooth mobility and loss. Porphyromonas gingivalis is considered the major etiological agent of this disease, having a plethora of virulence factors, including, lipopolysaccharides (LPS), hemolysins, and proteinases. Antimicrobial peptides are one of the main components of the innate immune response that inhibit the growth of P. gingivalis. The aim of this study was to analyze the antimicrobial activity of cystatin C and to assess the effect on the inflammatory and anti-inflammatory cytokines, the production of reactive oxygen species, and in the release of nitric oxide by human gingival fibroblasts incubated with P. gingivalis in the presence and absence of cystatin C.

Methods: P. gingivalis ATCC 33277 was exposed to cystatin C for 24h and co-cultured with human gingival fibroblasts (HGFs) ATCC CRL-2014. The effect of cystatin on growth of P. gingivalis and HGFs was evaluated. Pro-inflammatory (TNFα, IL-1β) and anti-inflammatory (IL-10) cytokines were determined by ELISA in the supernatants of HGFs incubated with P. gingivalis exposed to cystatin C. Additionally, nitrites and reactive oxygen species (ROS) production were evaluated.

Results: Cystatin Cinhibited the growth of P. gingivalis without affecting HGFs. Incubation of HGFs with P. gingivalis led to a significant increase of TNF-α and IL-1β. In contrast, HGFs incubated with P. gingivalis exposed to cystatin C showed a decreased production of both cytokines, whereas IL-10 was enhanced. Incubation of HGFs with P. gingivalis led to an increase of nitric oxide (NO) and ROS production, which was reduced in the presence of the peptide.

Conclusions: Cystatin C inhibits the growth of P. gingivalis and decreases the inflammatory cytokines, ROS, and NO production during infection of HGFs with P. gingivalis. Knowledge on the antimicrobial and immunomodulatory properties of cystatin C could aid in the design of new therapeutic approaches to facilitate the elimination of this bacterium to improve the treatment of periodontal disease.

Keywords: Cystatin C; Cytokines; Human gingival fibroblasts; Nitric oxide; Porphyromonas gingivalis; Reactive Oxigen Species.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Anti-Infective Agents* / pharmacology
  • Anti-Inflammatory Agents / pharmacology
  • Cystatin C / pharmacology
  • Cytokines / pharmacology
  • Fibroblasts
  • Humans
  • Interleukin-10 / pharmacology
  • Nitric Oxide / pharmacology
  • Periodontal Diseases*
  • Porphyromonas gingivalis
  • Reactive Oxygen Species / pharmacology

Substances

  • Interleukin-10
  • Reactive Oxygen Species
  • Cystatin C
  • Nitric Oxide
  • Cytokines
  • Anti-Inflammatory Agents
  • Anti-Infective Agents

Grants and funding

This work was supported by grant #IN218419 from PAPITT, DGAPA, UNAM, Mexico City, and by Universidad Anahuac México Campus Norte. Blanca Esther Blancas-Luciano is supported by CONACYT grant #424031 for her doctoral studies. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.