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  • TNF-α suppression and osteoprotegerin overexpression inhibits wear debris-induced inflammation and osteoclastogenesis in vitro.

TNF-α suppression and osteoprotegerin overexpression inhibits wear debris-induced inflammation and osteoclastogenesis in vitro.

The International journal of artificial organs (2015-11-07)
Li Peng, Haowei Zhang, Yongfei Zhou, Wenbo Li, Peng Jiang, Yunge Zhang, Keguan Song
ABSTRACT

Periprosthetic osteolysis, involving RANK/RANKL/osteoprotegerin (OPG) and TNF-α/NFκB signaling, contributes to bone resorption and inflammation. We constructed lentivirus vectors to inhibit TNF-α and enhance OPG expression and assessed their impacts on wear debris-induced inflammation and osteoclastogenesis in an osteoclast/osteoblast coculture system. We transduced mouse osteoblastic MC3T3-E1 cells with Lenti-negative control (Lenti-NC), Lenti-OPG or Lenti-siTNFα-OPG, and murine macrophage/monocyte RAW264.7 cells with Lenti-NC, Lenti-TNF-α siRNA or Lenti-siTNFα-OPG. Then, TNF-α and OPG protein levels were evaluated by enzyme-linked immunosorbent assay. We cocultured transduced MC3T3-E1 and RAW264.7 cells in transwell chambers in the presence of 0.1 mg/mL Ti particles to investigate the capacity of TNF-α inhibition to reduce wear debris-induced inflammation. We also assessed mRNA levels TNF-α, IL-1β, IL-6 and OPG by RT-PCR as well as osteoclastogenesis by tartrate-resistant acid phosphatase. Lenti-siTNFα-OPG ameliorated Ti-particle-induced expression of TNF-α, IL-1β, IL-6 in MC3T3-E1/RAW264.7 cocultures, while enhancing mRNA and protein levels of OPG, and reducing the fraction of tartrate-resistant acid phosphatase (TRAP)+ cells. Lenti-siTNFα-OPG can inhibit the wear debris-induced inflammatory responses and osteoclastogenesis in vitro, and may represent a promising therapeutic candidate for the treatment or prevention of wear particle-induced osteolysis.

MATERIALS
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Brand
Product Description

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Ser-Phe-Leu-Leu-Arg-Asn-Pro-Asn-Asp-Lys-Tyr-Glu-Pro-Phe, ≥97% (HPLC)
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MISSION® esiRNA, targeting mouse Tnf