| BMC Oral Health | |
| Palatal mucosa derived fibroblasts present an adaptive behavior regarding cytokine secretion when grafted onto the gingival margin | |
| Sebastião Luiz Aguiar Greghi1  Adriana Campos Passanezi Sant’Ana1  Maria Lúcia Rubo de Rezende1  Carla Andreotti Damante1  Carlos Ferreira Santos2  Thiago José Dionísio2  Carla Renata Sipert2  Ana Carolina Faria Morandini2  Fabíola Pontes Azevedo1  | |
| [1] Department of Prosthodontics, Bauru School of Dentistry, University of São Paulo, Bauru, São Paulo, Brazil;Department of Biological Sciences, Bauru School of Dentistry, University of São Paulo, Bauru, São Paulo, Brazil | |
| 关键词: Inflammation; Chemokine; Cytokines; Gingival fibroblasts; Periodontitis; | |
| Others : 1118683 DOI : 10.1186/1472-6831-14-21 |
|
| received in 2013-12-13, accepted in 2014-03-07, 发布年份 2014 | |
PDF
|
|
【 摘 要 】
Background
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.
Methods
Biopsies 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).
Results
Fibroblasts 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.
Conclusion
Fibroblasts 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.
【 授权许可】
2014 Azevedo et al.; licensee BioMed Central Ltd.
【 预 览 】
| Files | Size | Format | View |
|---|---|---|---|
| 20150207025526240.pdf | 1123KB | ||
| Figure 6. | 55KB | Image | |
| Figure 5. | 58KB | Image | |
| Figure 4. | 58KB | Image | |
| Figure 3. | 57KB | Image | |
| Figure 2. | 46KB | Image | |
| Figure 1. | 62KB | Image |
【 图 表 】
Figure 1.
Figure 2.
Figure 3.
Figure 4.
Figure 5.
Figure 6.
【 参考文献 】
- [1]Mariotti A: Dental plaque-induced gingival diseases. Ann Periodontol 1999, 4:7-19.
- [2]Löe H, Anerud A, Boysen H: The natural history of periodontal disease in man: prevalence, severity, and extent of gingival recession. J Periodontol 1992, 63:489-495.
- [3]Kassab MM, Cohen RE: The etiology and prevalence of gingival recession. J Am Dent Assoc 2003, 134:220-225.
- [4]Toker H, Ozdemir H: Gingival recession: epidemiology and risk indicators in a university dental hospital in Turkey. Int J Dent Hyg 2009, 7:115-120.
- [5]Greenwell H, Fiorellini J, Giannobile W, Offenbacher S, Salkin L, Townsend C, Sheridan P, Genco R: Oral reconstructive and corrective considerations in periodontal therapy. J Periodontol 2005, 76:1588-1600.
- [6]Marquez IC: The role of keratinized tissue and attached gingiva in maintaining periodontal/peri-implant health. Gen Dent 2004, 52:74-79.
- [7]Brunner J, Scheres N, El Idrissi NB, Deng DM, Laine ML, van Winkelhoff AJ, Crielaard W: The capsule of Porphyromonas gingivalis reduces the immune response of human gingival fibroblasts. BMC Microbiol 2010, 10:1-11. BioMed Central Full Text
- [8]Morandini AC, Sipert CR, Ramos-Junior ES, Brozoski DT, Santos CF: Periodontal ligament and gingival fibroblasts participate in the production of TGF-β, interleukin (IL)-8 and IL-10. Braz Oral Res 2011, 25:157-162.
- [9]Morandini AC, Sipert CR, Gasparoto TH, Greghi SL, Passanezi E, Rezende ML, Sant'ana AP, Campanelli AP, Garlet GP, Santos CF: Differential production of macrophage inflammatory protein 1-α, stromal-derived factor-1, and IL-6 by human cultured periodontal ligament and gingival fibroblasts challenged with lipopolyssaccharide from P. gingivalis. J Periodontol 2010, 81:310-317.
- [10]Morandini AC, Souza PPC, Ramos-Junior ES, Costa CAS, Santos CF: MyD88 or TRAM knockdown regulates interleukin (IL)-6, IL-8 and CXCL12 mRNA expression in human gingival and periodontal ligament fibroblasts. J Periodontol 2013, 84:1353-1360.
- [11]Morandini ACF, Souza PPC, Ramos-Junior ES, Brozoski DT, Sipert CR, Souza Costa CA, Santos CF: Toll-like receptor 2 knockdown modulates interleukin (IL)-6 and IL-8 but not stromal derived factor-1 (SDF-1/CXCL12) in human periodontal ligament and gingival fibroblasts. J Periodontol 2013, 84:535-544.
- [12]Bartold PM, Walsh LJ, Narayanan AS: Molecular and cell biology of the gingiva. Periodontol 2000 2000, 24:28-35.
- [13]Ara T, Kurata K, Hirai K, Uchihashi T, Uematsu T, Imamura Y, Furusawa K, Kurihara S, Wang PL: Human gingival fibroblasts are critical in sustaining inflammation in periodontal disease. J Periodontal Res 2009, 44:21-27.
- [14]Scheres N, Laine ML, de Vries TJ, Everts V, van Winkelhoff AJ: Gingival and periodontal ligament fibroblasts differ in their inflammatory response to viable Porphyromonas gingivalis. J Periodontal Res 2010, 45:262-270.
- [15]Herath TD, Wang Y, Seneviratne CJ, Lu Q, Darveau RP, Wang CY, Jin L: Porphyromonas gingivalis lipopolysaccharide lipid A heterogeneity differentially modulates the expression of IL-6 and IL-8 in human gingival fibroblasts. J Clin Periodontol 2011, 38:694-701.
- [16]Garlet GP, Martins W Jr, Ferreira BR, Milanezi CM, Silva JS: Patterns of chemokines and chemokine receptors expression in different forms of human periodontal disease. J Periodontal Res 2003, 38:210-217.
- [17]Arancibia R, Oyarzún A, Silva D, Tobar N, Martínez J, Smith PC: Tumor necrosis factor-α inhibits transforming growth factor-β-stimulated myofibroblastic differentiation and extracellular matrix production in human gingival fibroblasts. J Periodontol 2013, 84:683-693.
- [18]Johnson RB, Serio FG, Dai X: Vascular endothelial growth factors and progression of periodontal diseases. J Periodontol 1999, 70:848-852.
- [19]Hosokawa Y, Hosokawa I, Ozaki K, Nakae H, Matsuo T: CXC chemokine ligand 16 in periodontal diseases: expression in diseased tissues and production by cytokine-stimulated human gingival fibroblasts. Clin Exp Immunol 2007, 149:146-154.
- [20]Hosokawa Y, Hosokawa I, Ozaki K, Nakae H, Matsuo T: Human gingival fibroblasts express functional chemokine receptor CXCR6. Clin Exp Immunol 2009, 156:413-418.
- [21]Sullivan HC, Atkins JH: Free autogenous gingival grafts. III. Utilization of grafts in the treatment of gingival recession. Periodontics 1968, 6:152-160.
- [22]Scheres N, Laine ML, Sipos PM, Bosch-Tijhof CJ, Crielaard W, de Vries TJ, Everts V: Periodontal ligament and gingival fibroblasts from periodontitis patients are more active in interaction with Porphyromonas gingivalis. J Periodontal Res 2011, 46:407-416.
- [23]Hosokawa Y, Hosokawa I, Ozaki K, Nakae H, Murakami K, Miyake Y, Matsuo T: CXCL12 and CXCR4 expression by human gingival fibroblasts in periodontal disease. Clin Exp Immunol 2005, 141:467-474.
- [24]Uehara A, Takada H: Functional TLRs and NODs in human gingival fibroblasts. J Dent Res 2007, 86:249-254.
- [25]Nishi H, Ohta K, Takechi M, Yoneda S, Hiraoka M, Kamata N: Wound healing effects of gingival fibroblasts cultured in animal-free medium. Oral Dis 2010, 16:438-444.
- [26]Agis H, Watzek G, Gruber R: Prolyl hydroxylase inhibitors increase the production of vascular endothelial growth factor by periodontal fibroblasts. J Periodontol Res 2012, 47:165-173.
- [27]Koka S, Reinhardt RA: Periodontal pathogen-related stimulation indicates unique phenotype of primary cultured human fibroblasts from gingiva and periodontal ligament: implications for oral health disease. J Prosthet Dent 1997, 77:191-196.
- [28]Ekhlassi S, Scruggs LY, Garza T, Montufar-Scolis D, Moretti AJ, Klein JR: Porphyromonas gingivalis lipopolysaccharide induces tumor necrosis factor-α and interleukin-6 (IL-6) secretion and CCL2 gene expression in mouse primary gingival cell lines: IL-6-driven activation of CCL2. J Periodontal Res 2008, 43:431-439.
- [29]Tamura M, Tokuda M, Nagaoka S, Takada H: Lipopolysaccharides of Bacteroides intermedius (Prevotella intermedia) and Bacteroides (Porphyromonas) gingivalis induce interleukin-8 gene expression in human gingival fibroblast cultures. Infect Immun 1992, 60:4932-4937.
- [30]Yamaji Y, Kubota T, Sasaguri K, Sato S, Suzuki Y, Kumada H, Umemoto T: Inflammatory cytokine gene expression in human periodontal ligament fibroblasts stimulated with bacterial lipopolysaccharides. Infect Immun 1995, 63:3576-3581.
- [31]Suthin K, Matsushita K, Machigashira M, Tatsuyama S, Imamura T, Torii M, Izumi Y: Enhanced expression of vascular endothelial growth factor by periodontal pathogens in gingival fibroblasts. J Periodontal Res 2003, 38:90-96.
- [32]Núñez MJ, Novío S, Balboa J, Seoane J, Suárez JA, Freire-Garabal M: Effects of resveratrol on expression of vascular endothelial growth factor in human gingival fibroblasts stimulated by periodontal pathogens. Acta Odontol Scand 2010, 68:239-247.
- [33]Abel S, Hundhausen C, Mentlein R, Schulte A, Berkhout TA, Broadway N, Hartmann D, Sedlacek R, Dietrich S, Muetze B, Schuster B, Kallen KJ, Saftig P, Rose-John S, Ludwig A: The transmembrane CXC-chemokine ligand 16 is induced by IFN-gamma and TNF-alpha and shed by the activity of the disintegrin-like metalloproteinase ADAM10. J Immunol 2004, 172:6362-6372.
PDF