Evaluation of Proinflammatory Response to Polymeric Materials Using a Macrophage Cell Line Genetically Tagged with a Luminescent Peptide
Tsuyoshi KIMURA, Hanako MAEDA, Moeko HAGIWARA, Yoshihide HASHIMOTO, Naoko NAKAMURA, Wataru NOMURA, Tadao TANABE, Mako KOBAYASHI, Masaya YAMAMOTO, Takahide MATSUSHIMA, Hiroshi ASAHARA, Akio KISHIDA
Vol. 13 (2024) p. 43-51
Investigation of biological response to materials is important in understanding their biocompatibility and cell-material interactions for biomaterial applications. Macrophages are important for early biological response. Responses of macrophages to materials have previously been investigated by quantitating inflammatory and anti-inflammatory cytokines using ELISA and RT-PCR assays, and by assessing phenotype changes using flow cytometry and immunohistochemistry. In this study, we developed a method to evaluate the proinflammatory response to polymeric materials using a macrophage cell line (THP-1) genetically tagged with a luminescent peptide (HiBiT). The gene for the luminescent peptide was inserted into IL-1β in THP-1 cells using the CRISPR/Cas9 system. Upon stimulation of HiBiT-tagged THP-1 cells with lipopolysaccharide, IL-1β secretion could be detected using highly sensitive measurement of luminescence as well as using ELISA and RT-PCR assays. We found that IL-1β production by HiBiT-tagged THP-1 cells differed in response to nylon, cellulose, and polytetrafluoroethylene. Moreover, the time course of IL-1β secretion also differed for these materials. These results indicate that IL-1β production over time in HiBiT-tagged THP-1 cells exposed to a material can be measured. We believe that this method for evaluation of proinflammatory response using genetically engineered macrophages would complement ELISA and RT-PCR in investigating cellular response to different materials.