In Vitro Regulation of CCL3 and CXCL12 by Bacterial By-products Is Dependent on Site of Origin of Human Oral Fibroblasts

Abstract

Introduction Production of chemokines by tissue resident cells is one of the main mechanisms involved in the inflammatory infiltrate formation during inflammation. The specific ability of fibroblasts from different oral tissues such as gingiva, periodontal ligament and dental pulp from permanent and deciduous teeth in producing the chemokines CCL3 and CXCL12 under stimulation by bacterial products commonly found in endodontic infections was investigated. Methods Cultures of fibroblasts from gingiva and periodontal ligament as well as from dental pulp from permanent and deciduous teeth were established by using an explant technique and stimulated with increasing concentrations of Escherichia coli lipopolysaccharide (EcLPS) and Enterococcus faecalis lipoteichoic acid (EfLTA) for 1, 6 and 24 hours. Supernatants were tested for CCL3 and CXCL12 by enzyme-linked immunosorbent assay. Results In general, CCL3 production was induced by EcLPS in the 4 fibroblast subtypes and by EfLTA in fibroblasts from gingiva and periodontal ligament. Constitutive CXCL12 synthesis decreased in all fibroblast subtypes especially under stimulation with EcLPS. Fibroblast from permanent deciduous teeth was the cell type presenting the most expressive reduction in CXCL12 release by both stimuli. On the basis of computational matching of CXCL12 mRNA with the microRNAs miR-141 and miR-200a, their expression was also investigated. Although detected in the fibroblasts, these molecules remained unaltered by bacterial by-product stimulation. Conclusions EcLPS and EfLTA induced the production of CCL3 and unbalanced the synthesis of CXCL12 in a manner dependent on the specific tissue origin of fibroblasts.

Angiotensin II Type 1 Receptor Knockdown Impairs Interleukin‐1β‐Induced Cytokines in Human Periodontal Fibroblasts

“…Cultured cells were characterized as fibroblasts by their morphology and positivity for fibroblast surface protein (FSP)‐1 as previously described 10 , 32 . Immunostaining was performed by using mouse antihuman FSP‐1 (1B10), ‡ followed by fluorescein‐conjugated antimouse immunoglobulin G § .…”

Expression of the stem cell factor in fibroblasts, endothelial cells, and macrophages in periapical tissues in human chronic periapical diseases

“…LPS in infected pulp tissues of pulp canal could activate LYMs and FBs producing pro-inflammatory cytokines such as TNF and IL-1 (Chang et al, 2005). Some studies have suggested that FBs could be activated to secret inflammatory cytokines, such as NLRP3 and IL-33, after stimulation of inflammation and participate in infiltration, cell transport, and tissues fibrosis in the tissue repair process (Liu and Liu, 2013;Sipert et al, 2014;Velickovic et al, 2016).…”

MicroRNA-146a and microRNA-155 show tissue-dependent expression in dental pulp, gingival and periodontal ligament fibroblasts in vitro

“…With written informed consent, teeth and gingival tissues were donated by patients undergoing lower third molar removal. Cultures of human fibroblasts from dental pulp, gingiva and periodontal ligament from the same donors (n = 3) (2 females and 1 male, 16-26 years old) were established by means of the explant technique as described elsewhere (5,7,23). Immediately after removal, the tissues were kept separately under aseptic conditions.…”

“…However, it has been clearly demonstrated that the biological behavior of fibroblasts is related to the type of connective tissue from which they originate (2)(3)(4). Such properties include membrane receptor expression (3) and cytokine release kinetics (2,3,(5)(6)(7).…”