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

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.

Cystatin C, Ammonia, and Bicarbonate Measurements in the Saliva of Pigs: Analytical Validation and Changes in S. suis Infection

“…It can inhibit the growth of bacteria, bind to the active sites of cysteine and thiol proteases, reduce their function, and decrease the production of mediators of inflammation and also ROS and NO in bacterial infection. Therefore, it has antimicrobial and immunomodulation properties [ 28 ]. Immunomodulation is specially focused on the immune innate response with modulation of cathepsin activity, antigen presentation, and synthesis of tumor necrosis factor-alpha and interleukin 10 [ 29 ].…”

Type 2 Cystatins and Their Roles in the Regulation of Human Immune Response and Cancer Progression

“…The anti-microbial mechanism of SD-type cystatins is not fully understood, but SD-type cystatins may be inhibiting proteolytic activity necessary for growth promotion in the bacteria [24]. This antimicrobial effect is also observed in human cystatin C and cystatins isolated from other species [25]. Given the conserved nature of this antimicrobial effect, type 2 cystatins may have originally served as antimicrobials [26].…”

Association between Serum Cys C and PTB Cavitation

“…Therefore, we speculate that Cys C may participate in PTB cavitation via additional mechanisms. Recent studies have shown that Cys C plays a crucial role in regulating antigen presentation, protecting the host from microbial invasion, and inhibiting viral replication and growth [20][21][22][23]. Elevated Cys C levels associated with high viral loads have been found in HIV patients [24].…”