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

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. 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 . 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 in the presence and absence of cystatin C.

METHODS

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 and HGFs was evaluated. Pro-inflammatory (TNF, IL-1) and anti-inflammatory (IL-10) cytokines were determined by ELISA in the supernatants of HGFs incubated with exposed to cystatin C. Additionally, nitrites and reactive oxygen species (ROS) production were evaluated.

RESULTS

Cystatin Cinhibited the growth of without affecting HGFs. Incubation of HGFs with led to a significant increase of TNF- and IL-1. In contrast, HGFs incubated with exposed to cystatin C showed a decreased production of both cytokines, whereas IL-10 was enhanced. Incubation of HGFs with 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 . 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.