Cytotoxicity and potential anti-inflammatory activity of velutin on RAW 264.7 cell line differentiation: Implications in periodontal bone loss.

OBJECTIVE

Hypoxia-inducible factor-1α (HIF-1α) has been implicated in periodontal tissue inflammation and possibly in osteoclast differentiation, while polyphenols are known to be anti-inflammatory natural compounds that are capable of regulating the NF-κB protein complex pathway. The objective of this study was to investigate cytotoxicity and HIF-1α expression through the NF-κB pathway by polyphenol velutin (Euterpe oleracea Mart.), found in the pulp of acai fruit, during inflammatory RAW 264.7 differentiation.

METHODS

RAW 264.7 mouse monocyte macrophage cells were stimulated with RANKL (30ng/mL) and Porphyromonas gingivalis lipopolysaccharide (1μg/mL). Cells were treated with various concentrations of velutin (0.5-2μM) to check for viability, morphology, osteoclast differentiation, and HIF-1α expression (Western blot).

RESULTS

Alamar blue cell viability assay showed no toxicity to RAW cells with the use of velutin in all concentrations tested (p>0.05). Velutin did not induce cell apoptosis based on caspase 3/7 assay (p>0.05). Fluorescence images stained by DAPI showed no alteration in the morphology of RAW cell nuclei (p>0.05) treated with velutin. TRAP assays demonstrated a dose-dependent reduction in osteoclast formation by velutin when compared with control (p<0.05). Velutin showed a reduction in HIF-1α expression related to IκB phosphorylation when compared with control (p<0.001).

CONCLUSIONS

At the tested concentrations, velutin was not cytotoxic to RAW 264.7 and differentiated cells. Velutin reduced osteoclast differentiation and downregulated HIF-1α through the NF-κB pathway.