Application of small interfering RNA for inhibition of lipopolysaccharide-induced osteoclast formation and cytokine stimulation

Journal of Endodontics
Farshid S FahidLarz S Spangberg

Abstract

RNA interference (RNAi) is a unique and powerful tool used for the study of gene function by suppressing its expression. Nuclear factor of activated T cells (NFATc1) is the most strongly induced transcription factor mediated by receptor activator for nuclear factor kappa B ligand stimulation and has shown to be a key regulator of osteoclastogenesis. To determine the application of small interfering RNA (siRNA) for inhibition of lipopolysaccharide (LPS)-induced cytokine stimulation and osteoclast formation, murine monocyte, RAW 264.7 cells as well as differentiated osteoclasts were transfected with NFATc1-specific siRNA and then stimulated with 100 ng/mL LPS. By using real-time polymerase chain reaction analysis and enzyme-linked immunosorbent assay, we confirmed that monocytes whose NFATc1 protein expression was silenced by using RNAi produced lower levels of inflammatory cytokines, fewer numbers evolved into mature osteoclasts, and osteoclasts expressed lower levels of osteoclast-specific gene markers such as tartrate-resistant acid phosphatase and cathepsin K. These results suggested that RNAi could be used to modulate the effects of LPS stimulation.

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Citations

Oct 15, 2013·Therapeutic Delivery·Yuwei Wang, David W Grainger
Nov 10, 2010·Expert Opinion on Biological Therapy·Qin ShiJulio C Fernandes
Sep 29, 2011·Advanced Drug Delivery Reviews·Yuwei Wang, David W Grainger
Jul 20, 2017·Journal of Biomaterials Science. Polymer Edition·Deniz Sezlev BilecenVasif Hasirci
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Jul 18, 2021·Inflammation Research : Official Journal of the European Histamine Research Society ... [et Al.]·Mouna FerbebouhJulio Fernandes

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