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Comparison of the cytotoxic and inflammatory responses of titanium particles with different methods for endotoxin removal in RAW264.7 macrophages

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Abstract

It is generally accepted that periprosthetic bone resorption is initiated through aseptic inflammation aggravated by wear particles that are generated from artificial joint. However, some studies have demonstrated that “endotoxin-free” wear particles are almost completely unable to stimulate the macrophage-mediated production of proinflammatory cytokines. Here, we compare the titanium particles with different methods of endotoxin removal. The results indicated that different titanium particle preparation dosages did not significantly change particle size, morphology, and chemical composition. But it could cause variations in the endotoxin concentration of titanium particles and inflammatory responses in RAW264.7 macrophages. The particles with higher endotoxin levels correlated with more extensive inflammatory responses. When testing endotoxins using the supernatant of particle suspensions, it would lead to false negative results compared with testing the particle themselves. And when using the particles themselves, all the particles should be removed by centrifugation to avoid particle interference before the absorbance value was determined. Therefore, we suggest that research concerning wear particles should completely describe the endotoxin testing process, including endotoxin removal from particles and the details of endotoxin testing. Moreover, future research should focus on the surface of wear particles (the potential role of adherent endotoxin) rather than the particles themselves.

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Acknowledgments

This work was supported by the National High Technology Research and Development Program of China (863 Program) (Grant No. 2006AA02A137), Program for the Shanghai Key Laboratory of Orthopaedic Implant (Grant No. 08DZ2230300) and the National Natural Science Foundation of China (Grant No. 81001529). We also thank Prof. Jiake Xu from the University of Western Australia for his donation of the RAW264.7 cells that were stably transfected with a luciferase reporter gene.

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Authors and Affiliations

  1. Department of Orthopaedic Surgery, Shanghai 9th People’s Hospital, School of Medicine, Shanghai Jiao Tong University, 639 Zhi Zao Ju Road, Shanghai, 200011, People’s Republic of China

    Huifeng Ding, Zhenan Zhu, Tingting Tang, Degang Yu, Bo Yu & Kerong Dai

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  1. Huifeng Ding
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Correspondence to Zhenan Zhu.

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Ding, H., Zhu, Z., Tang, T. et al. Comparison of the cytotoxic and inflammatory responses of titanium particles with different methods for endotoxin removal in RAW264.7 macrophages. J Mater Sci: Mater Med 23, 1055–1062 (2012). https://doi.org/10.1007/s10856-012-4574-x

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