Heat-killed Enterococcus faecalis Alters Nitric Oxide and CXCL12 Production but not CXCL8 and CCL3 Production by Cultured Human Dental Pulp Fibroblasts

Sipert, Carla Renata; Moraes, Ivaldo Gomes de; Bernardinelli, Norberti; Garcia, Roberto B.; Bramante, Clóvis Monteiro; Gasparoto, Thaís Helena; Figueira, Eduardo Aleixo; Dionísio, Thiago José; Campanelli, Ana Paula; Oliveira, Sandra Helena Penha de; Cunha, Fernando; Santos, Carlos

doi:10.1016/j.joen.2009.10.014

Cited by 29 publications

(32 citation statements)

References 40 publications

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“…E. faecalis attached in the form of cocci or chains, indicated with a white arrow, to the cells (LT-2) stimulated a high proliferative subpopulation of quiescent cells, possibly through NO-mediated mechanisms [34]. In contrary, viability and proliferation assays did not disclose any cell number changes when human dental pulp fibroblasts were seeded with whole heat-inactivated E. faecalis [35]. The results of the present study contribute to the above conflicting published data.…”

Section: Discussionmentioning

confidence: 41%

Enterococcus faecalis affects the proliferation and differentiation of ovine osteoblast-like cells

et al. 2011

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Enterococcus faecalis (E. faecalis) is a Gram-positive bacterium, mostly recovered from root-filled teeth with persistent periapical lesions. Bacterial contamination of root canals inevitably results in interaction between E. faecalis and periapical tissues during the dynamic process of periapical inflammation. This study investigated the impact of heat-inactivated endodontic E. faecalis on the proliferation and the differentiation of ovine osteoblast-like cells, in an attempt to elucidate its putative enhanced pathogenicity mechanisms. Therefore, two different concentrations of a heat-inactivated endodontic E. faecalis isolate (2 × 10(6) or 2 × 10(8) CFU/ml) were incubated with ovine osteoblast-like cells for 7 and 14 days, respectively. Cells without antigen served as control. The effects of antigen on cell growth were evaluated by a proliferation assay (EZ4U). Furthermore, the assessment of alkaline phosphatase (ALP) activity, calcium deposition, and osteocalcin (OCN) gene expression through quantitative real-time PCR determined the degree of osteogenic cell differentiation. Scanning electron microscopy (SEM) was also performed to detect alterations in cell morphology. Interestingly, although highly concentrated E. faecalis increased cellular reproduction after 14 days, ALP activity and OCN gene expression decreased in an antigen concentration-dependent and incubation time-independent way. SEM images revealed E. faecalis adhesion on cells, a fact that might contribute to its virulence. These results suggest that E. faecalis stimulated cell multiplication, whereas it likely restrained cell differentiation of ovine osteoblast-like cells. In conclusion, the presence of E. faecalis in root canals may negatively affect periapical new bone formation, and thus, the healing of periapical lesions.

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Section: Discussionmentioning

confidence: 41%

Enterococcus faecalis affects the proliferation and differentiation of ovine osteoblast-like cells

et al. 2011

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“…Primary culture of periodontal ligament fibroblasts was established using an explant technique as previously described (11,12). In brief, the periodontal ligament of a third molar was carefully removed and placed in Dulbecco's Modified Eagle's Medium (DMEM -Gibco/Invitrogen Corporation, CA, USA) under aseptic conditions for cell cultivation.…”

Section: Cell Culturementioning

confidence: 99%

“…For this purpose, the Griess method was employed (12). Aliquots of 50 µL of each cell supernatant were added to a 96-well plate in duplicate.…”

Section: Nitric Oxide (No) Measurementmentioning

confidence: 99%

In Vitro Cytotoxicity of White MTA, MTA Fillapex® and Portland Cement on Human Periodontal Ligament Fibroblasts

et al. 2013

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The aim of this study was to compare the in vitro cytotoxicity of white mineral trioxide aggregate (MTA), MTA Fillapex ® and Portland cement (PC) on human cultured periodontal ligament fibroblasts. Periodontal ligament fibroblast culture was established and the cells were used for cytotoxic tests after the fourth passage. Cell density was set at 1.25 x10 4 cells/well in 96-well plates. Endodontic material extracts were prepared by placing sealer/ cement specimens (5x3mm) in 1mL of culture medium for 72 h. The extracts were then serially two-fold diluted and inserted into the cell-seeded wells for 24, 48 and 72 h. MTT assay was employed for analysis of cell viability. Cell supernatants were tested for nitric oxide using the Griess reagent system. MTA presented cytotoxic effect in undiluted extracts at 24 and 72 h. MTA Fillapex ® presented the highest cytotoxic levels with important cell viability reduction for pure extracts and at ½ and ¼ dilutions. In this study, PC did not induce alterations in fibroblast viability. Nitric oxide was detected in extract-treated cell supernatants and also in the extracts only, suggesting presence of nitrite in the soluble content of the tested materials. In the present study, MTA Fillapex displayed the highest cytotoxic effect on periodontal ligament fibroblasts followed by white MTA and PC.

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“…In other words, there is evidence that SCF production occurs via tumor necrosis factor a and IL-6 production, but further experiments should be accomplished in order to address which cytokines and/or chemokines are involved in the process that mediates SCF production by ODs. The stromal cellderived factor 1a (SDF-1a) may be an important chemokine involved in the migration and differentiation of tissue-committed stem cells to injury sites under some pathological conditions (26); however, the relationship among SCFs, ODs, and LPS from E. coli is under investigation, but data from the literature described the regulation of the SDF-1a-chemokine (C-X-C motif) receptor 4 (CXCR4) axis in human dental pulp by LPS from E. coli or heat-killed Enterococcus faecalis, showing that LPS regulated SDF-1a production and CXCR4 expression in dental pulp cells (27)(28)(29).…”

Section: Discussionmentioning

confidence: 99%

Lipopolysaccharide-induced Stem Cell Factor Messenger RNA Expression and Production in Odontoblast-like Cells

Oliveira

Santos

2012

Journal of Endodontics

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Heat-killed Enterococcus faecalis Alters Nitric Oxide and CXCL12 Production but not CXCL8 and CCL3 Production by Cultured Human Dental Pulp Fibroblasts

Cited by 29 publications

References 40 publications

Enterococcus faecalis affects the proliferation and differentiation of ovine osteoblast-like cells

Enterococcus faecalis affects the proliferation and differentiation of ovine osteoblast-like cells

In Vitro Cytotoxicity of White MTA, MTA Fillapex® and Portland Cement on Human Periodontal Ligament Fibroblasts

Lipopolysaccharide-induced Stem Cell Factor Messenger RNA Expression and Production in Odontoblast-like Cells

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