Abstract
BackgroundConsidering that grafted gingival tissue might have to be adapted to the receptor area and that fibroblasts have the ability to respond to bacterial stimuli through the release of various cytokines, this study investigated whether fibroblasts from the palatal mucosa behave differently when grafted onto the gingival margin regarding cytokine secretion.MethodsBiopsies from the palatal mucosa were collected at the time of free gingival graft surgery, and after four months re-collection was performed upon surgery for root coverage. Fibroblasts were isolated by the explant technique, cultured and stimulated with Porphyromonas gingivalis (Pg) and Escherichia coli (Ec) LPS for 24 or 48 h for comparative evaluation of the secretion of cytokines and chemokines, such as IL-6, IL-8/CXCL8, MIP-1α/CCL3, TGF-β, VEGF and CXCL16. Unstimulated cells were used as the control group. Cells were tested for viability through MTT assay, and secretion of cytokines and chemokines was evaluated in the cell supernatants by Enzyme-Linked Immunosorbent Assay (ELISA).ResultsFibroblasts from the palatal mucosa maintained the same secretion pattern of IL-6 when grafted onto the gingival margin. On the contrary, fibroblasts from the marginal gingival graft showed increased secretion of IL-8/CXCL8 even in the absence of stimulation. Interestingly, MIP-1α/CCL3 secretion by fibroblasts from the marginal gingival graft was significantly increased after 48 hours of stimulation with Pg LPS and after 24 h with Ec LPS. Only fibroblasts from the marginal gingival graft showed secretion of TGF-β. VEGF and CXCL16 secretion were not detected by both subsets of fibroblasts.ConclusionFibroblasts from the palatal mucosa seem to be adapted to local conditions of the site microenvironment when grafted onto the gingival marginal area. This evidence supports the effective participation of fibroblasts in the homeostasis of the marginal periodontium through secretion modulation of important inflammatory mediators.
Highlights
Considering that grafted gingival tissue might have to be adapted to the receptor area and that fibroblasts have the ability to respond to bacterial stimuli through the release of various cytokines, this study investigated whether fibroblasts from the palatal mucosa behave differently when grafted onto the gingival margin regarding cytokine secretion
It has been shown that fibroblasts, besides being the most important cells in the maintenance and remodeling of connective tissue extracellular matrix, are involved in immune and inflammatory host response in periodontal disease, as they are the predominant structural cells found in periodontal infections by anaerobic bacteria such as Porphyromonas gingivalis (Pg) and can be stimulated to release inflammatory cytokines upon Pg and its subproducts challenge [7,8,9,10,11]
For IL-6 and IL-8/CXCL8, a well-known chemokine required for neutrophils recruitment, it was reported that these cells are able to secrete huge amounts when stimulated by Pg LPS [8,13,14,15]
Summary
Considering that grafted gingival tissue might have to be adapted to the receptor area and that fibroblasts have the ability to respond to bacterial stimuli through the release of various cytokines, this study investigated whether fibroblasts from the palatal mucosa behave differently when grafted onto the gingival margin regarding cytokine secretion. VEGF is another cytokine related to the etiology of periodontal disease due to the ability to increase vascular permeability that contributes to the dissemination of inflammation, allowing inflammatory mediator release by cells in the inflamed tissue and enabling the formation of new blood vessels [18]. To the best of our knowledge, no previous work has addressed the secretion profile of the aforementioned cytokines comparing the fibroblasts from palatal mucosa (usually used as an autogenous donor site for gingival graft procedures) with cells originated from marginal gingival area
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