Adenovirus-mediated siRNA targeting TNF-α and overexpression of bone morphogenetic protein-2 promotes early osteoblast differentiation on a cell model of Ti particle-induced inflammatory response in vitro

Abstract

Wear particles are phagocytosed by macrophages and other inflammatory cells, resulting in cellular activation and release of proinflammatory factors, which cause periprosthetic osteolysis and subsequent aseptic loosening, the most common causes of total joint arthroplasty failure. During this pathological process, tumor necrosis factor-alpha (TNF-α) plays an important role in wear-particle-induced osteolysis. In this study, recombination adenovirus (Ad) vectors carrying both target genes [TNF-α small interfering RNA (TNF-α-siRNA) and bone morphogenetic protein 2 (BMP-2)] were synthesized and transfected into RAW264.7 macrophages and pro-osteoblastic MC3T3-E1 cells, respectively. The target gene BMP-2, expressed on pro-osteoblastic MC3T3-E1 cells and silenced by the TNF-α gene on cells, was treated with titanium (Ti) particles that were assessed by real-time PCR and Western blot. We showed that recombinant adenovirus (Ad-siTNFα-BMP-2) can induce osteoblast differentiation when treated with conditioned medium (CM) containing RAW264.7 macrophages challenged with a combination of Ti particles and Ad-siTNFα-BMP-2 (Ti-ad CM) assessed by alkaline phosphatase activity. The receptor activator of nuclear factor-κB ligand was downregulated in pro-osteoblastic MC3T3-E1 cells treated with Ti-ad CM in comparison with conditioned medium of RAW264.7 macrophages challenged with Ti particles (Ti CM). We suggest that Ad-siTNFα-BMP-2 induced osteoblast differentiation and inhibited osteoclastogenesis on a cell model of a Ti particle-induced inflammatory response, which may provide a novel approach for the treatment of periprosthetic osteolysis.

The Effects of Biomaterial Implant Wear Debris on Osteoblasts

“…The communication between macrophages and osteoblasts during aseptic loosening has been studied in direct or indirect co-culture models. Upon particle stimulation, co-cultured macrophages secrete soluble factors such as TNFα, IL-6, IL-1β, and GM-CSF (Horowitz and Gonzales, 1996;Rodrigo et al, 2002;Vallés et al, 2008a;Guo et al, 2013) and stimulate the release of osteoblastic IL-6, PGE2, M-CSF, GM-CSF, MCP-1, and RANKL (Horowitz and Purdon, 1995;Haynes et al, 1997;Rodrigo et al, 2006;Vallés et al, 2008a). These inflammatory mediators lead to further macrophage recruitment, increased osteoclastogenesis, and suppression of osteoblast function (Horowitz et al, 1994;Horowitz and Purdon, 1995;Gallo et al, 2013;Guo et al, 2013).…”

“…Upon particle stimulation, co-cultured macrophages secrete soluble factors such as TNFα, IL-6, IL-1β, and GM-CSF (Horowitz and Gonzales, 1996;Rodrigo et al, 2002;Vallés et al, 2008a;Guo et al, 2013) and stimulate the release of osteoblastic IL-6, PGE2, M-CSF, GM-CSF, MCP-1, and RANKL (Horowitz and Purdon, 1995;Haynes et al, 1997;Rodrigo et al, 2006;Vallés et al, 2008a). These inflammatory mediators lead to further macrophage recruitment, increased osteoclastogenesis, and suppression of osteoblast function (Horowitz et al, 1994;Horowitz and Purdon, 1995;Gallo et al, 2013;Guo et al, 2013). In an indirect co-culture model, reduction of the TNFα level using neutralizing TNFα antibodies or TNFα-siRNA in the conditioned medium from particle-activated murine macrophages (J774/RAW 264.7) resulted in down-regulation of IL-6, PGE2, and GM-CSF in murine osteoblastic cells (MC3T3-E1) (Horowitz and Purdon, 1995;Guo et al, 2013).…”

“…These inflammatory mediators lead to further macrophage recruitment, increased osteoclastogenesis, and suppression of osteoblast function (Horowitz et al, 1994;Horowitz and Purdon, 1995;Gallo et al, 2013;Guo et al, 2013). In an indirect co-culture model, reduction of the TNFα level using neutralizing TNFα antibodies or TNFα-siRNA in the conditioned medium from particle-activated murine macrophages (J774/RAW 264.7) resulted in down-regulation of IL-6, PGE2, and GM-CSF in murine osteoblastic cells (MC3T3-E1) (Horowitz and Purdon, 1995;Guo et al, 2013). Similarly, TNFα and IL-1β antibody treatment induced similar effects in a THP-1 and human osteoblast co-culture model under challenge with TiO 2 rutile and pure Ti wear particles (Vallés et al, 2008a).…”

A bone-resorption surface-targeting nanoparticle to deliver anti-miR214 for osteoporosis therapy

“…8 To enhance bone-formation ability, drugs are delivered to the bone-formation area or osteoblasts. [9][10][11] To inhibit the function of osteoclasts efficiently, the delivery system should preferentially deliver drug to the bone-resorption area or osteoclasts. 12,13 Common medications used for osteoclast targeting OP treatments are bisphosphonates or modified bisphosphonates.…”

Inhibitory effect of quercetin on titanium particle induced endoplasmic reticulum stress related apoptosis and in vivo osteolysis

“…Total joint replacement (TJR), which is by the implantation of a permanent in-dwelling artificial prosthesis, is a highly successful procedure for promoting the agility in patients with joint dysfunction [ 1 ]. It has been proved that titanium (Ti) components have been widely used for joint replacement [ 2 ]. However, aseptic loosening due to periprosthetic osteolysis, induced by the adverse biological responses to wear particles, can damage the efficacy and longevity of the prosthetic components [ 3 , 4 ].…”