Expression of complement factor C1r and C1s in human gingival fibroblasts

Sammanfattning: Background: Periodontitis is an inflammatory condition rendering in degradation of tooth supporting tissue. In the inflammatory process cytokines, amongst others TNF-a, IL-1b and IL-6 play an important role in regulating the immune response. Periodontal Ehlers Danlos syndrome (pEDS) is a rare connective tissue disorder characterized by distinct oral manifestations such as periodontitis. Studies has shown that individuals with pEDS exhibit a mutation in the complement factors C1r and C1s. Aim: The aim of this study was to analyze the gene expression of C1r and C1s in human gingival fibroblasts and to investigate if the expression of these genes’ changes during inflammatory conditions. Methods: Human gingival fibroblasts (hGFBs) were cultured with TNF-a or IL-1b for 24h. RNA isolation, extraction and complementary DNA synthesis was made at the timepoint of 6h and 24h. An qRT-PCR analyses of the genes C1r, C1s and IL-6 were performed and the relative gene expression of C1r, C1s and IL-6 were calculated. Results: The expression of C1r and C1s increased after 24h in TNF-a and IL-1b compared to control by measuring the relative fold change of the genes by culturing fibroblast over time. The result was taken from three parallel samples and the result was significant. Conclusion: The findings point to a link between the inflammatory classical complement pathway and the connective tissue homeostasis. Hopefully, our findings will be one piece of the puzzle regarding the molecular events rendering in pEDS and thereby a potential treatment target.