In vitro activation of human fibroblasts by retrieved titanium alloy wear debris

Abstract

Titanium-aluminum-vanadium wear particles isolated from the soft-issue membrane of a failed total hip arthroplasty were added to human fibroblasts in cell culture. The cellular response to particle challenge was determined by assaying for levels of interleukin-1 beta, interleukin-6, tumor necrosis factor-alpha, prostaglandin E2, basic fibroblast growth factor, platelet-derived growth factor-AB, and transforming growth factor-beta. Collagenase and gelatinase activities were analyzed by zymography and [3H]collagen degradation. Cell viability was assessed by measuring the uptake of [3H]thymidine. Over the range of particle concentrations tested, cell viability, as demonstrated by [3H]thymidine uptake, remained unaffected. Fibroblasts exhibited a dose-dependent release of interleukin-6 in response to exposure to titanium-aluminum-vanadium particles. At 6 and 48 hours, the highest concentration of titanium alloy particles (0.189% [vol/vol]) resulted in 7-fold and 16-fold increases in interleukin-6 release, respectively, when compared with negative controls. Neither interleukin-1 beta nor tumor necrosis factor-alpha was detected in the culture medium at any particle concentration tested for both dermal and foreskin fibroblasts. The pattern of prostaglandin E2 release by fibroblasts mirrored the pattern of interleukin-6 release. Fibroblasts exposed to the highest concentration of titanium alloy particles showed an increase in collagenase activity, starting at 12 hours. When medium samples were treated with amino phenylmercuric acetate to activate latent enzymes, a statistically significant increase in collagenase activity was observed as early as 6 hours (p < 0.001). Substrate gel analysis of medium from fibroblasts stimulated by high particle concentrations also showed an increase in gelatinolytic activity when compared with unstimulated controls. Analysis of medium samples for growth factors showed an increase in basic fibroblast growth factor at low particle concentrations, beginning at 12 hours. Levels of platelet-derived growth factor-AB and transforming growth factor-beta were not detectable in the controls or at any particle concentration tested. The results of this study showed that fibroblasts exposed to titanium alloy wear particles become activated and release proinflammatory mediators that influence bone metabolism. These data support the hypothesis that direct activation of fibroblasts by particulate wear may play a role in particle-mediated osteolysis. Fibroblast activation coupled with the biologic response of macrophages to wear debris in the loosening membrane may have a synergistic effect on pathologic bone resorption.

Cytokine receptor profile of arthroplasty macrophages, foreign body giant cells and mature osteoclasts

“…With regard to aseptic loosening, IL-1, TNF and IL-6 have all been identified in periprosthetic tissues surrounding loose implants (Chiba et al 1994, Horikoshi et al 1994. Several studies have shown that these cytokines are released from wear particle-stimulated macrophages and fibroblastic cells (Glant et al 1993, Haynes et al 1993, Manlapaz et al 1996. Strong expression of gp130 was shown on arthroplasty macrophages, FBGCs and osteoclasts in this study.…”

“…Cytokines such as interleukin-l( (IL-l), tumor necrosis factor ( (TNF) and interleukin-6 (IL-6) are potent stimulators of osteoclastic bone resorption (Bertolini et al 1986, Gowen and Mundy 1986, Black et al 1991 and have been identified in the arthroplasty pseudomembrane that surrounds failed implants (Chiba et al 1994, Horikoshi et al 1994. In vivo and in vitro studies have shown that these cytokines are released from wear particle-stimulated macrophages and fibroblastic cells (Glant et al 1993, Haynes et al 1993, Jiranek et al 1993, Manlapaz et al 1996. The osteoclast is a highly specialized multinucleated cell which is responsible for bone resorption.…”

Effects of hydroxyapatite participate debris on the production of cytokines and proteases in human fibroblasts

“…They are abundant in the periprosthetic membranes that surround failed implants, and have been shown to phagocytize particles and respond by increasing the secretion of cytokines and proteases that are associated with enhanced bone resorption and the degradation of extracellular bone matrix [32,34].…”

How Has the Introduction of New Bearing Surfaces Altered the Biological Reactions to Byproducts of Wear and Modularity?

“…Volume 472, Number 12, December 2014 Biological Reactions to Byproducts of Wear 3701 periprosthetic environment and their possible contribution to inflammatory osteolysis [12,47,53,62,76]. Other cells of potential interest in the pathology of debris-induced osteolysis include osteoclasts [28,58,60], osteoblasts [71], lymphocytes [7], and mesenchymal cells [62].…”