![]() ![]() Reverse transcription of 500 ng of RNA to cDNA were performed via a high-capacity reverse transcriptase kit according to the manufacturer's instructions. Isolated RNA concentration was subsequently determined using a NanoDrop spectrophotometer. RNA were isolated from 8 × 106 cells using the RNeasy Mini Kit according to the manufacturer's instructions. After the incubation period at 37☌, cells were washed with sterile PBS and fixed for 30 minutes with 2% paraformaldehyde (PFA) for immunolabeling, or harvested with TRIzol lysis reagent for RNA assessment, respectively. For the cytokine challenge study, cells were exposed for 6 hours to 20 ng/ml IFN-γ and 100 ng/mL LPS, washed and then placed in 10% FBS 1% P/S DMEM for 24 hours. Mature macrophages were exposed to the following treatments for 24 hours: complete media (M0 control), 20 ng/ml IFN-γ and 100 ng/mL lipopolysaccharide LPS (M1 control), 20 ng/mL interleukin IL-4 (M2 control). ![]() It was hypothesized that IL-1β, IL-6, and TNF-α cytokine expression levels results would be different for PEEK and a novel zeolite filled PEEK composite (ZFuze, DiFusion Technologies). The combination of results from three separate kinds of studies would potentially weave a better understanding of PEEK fibrous encapsulation around retrieved implanted PEEK devices. The cell studies in combination with larger animal studies help understand the importance of appropriate choice in materials. Studies involving macrophages facilitate the understanding behind the mechanism of fibrous capsule formation anecdotally reported with PEEK implants. Based on the current understanding of the immune system, both the innate and adaptive components, a combination of tests on current design materials in spinal implant devices may help understand potential clinical complications and thus reduce the potential adverse events associated with specific design materials. Osteoimmunology, the study of the relationship between the musculoskeletal system and immune system, has emerged as an important consideration in biomaterial research. ![]()
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