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.