The Effect of Neutrophil-Derived Products on the Function of Leukocytes Obtained after Titanium Implantation in the Ovine Model

SIMPLE SUMMARY: Titanium is one of the most commonly used biomaterials for implantation as a part of the orthopedic procedures. However, this biomaterial can cause an excessive inflammatory response, even leading to rejection of the implant. Therefore, the aim of our study was to assess the overall organism response after insertion of Ti implant and the activity of neutrophils and monocyte-derived macrophages (MDM), to evaluate the possible negative effect of this biomaterial on the host cells. Our study revealed that insertion of the Ti implant did not evoke systemic inflammatory response or activation of leukocytes. Additionally, we evaluated the activity of neutrophils and MDM after stimulation with autologous neutrophil products, namely, antimicrobial neutrophil extract and neutrophil degranulation product as two potential regulators of inflammatory response. Antimicrobial neutrophil extract appeared to be a factor causing the decrease of secretory neutrophil response and polarization of MDM towards pro-resolving phenotype, whereas the neutrophil degranulation product acted as pro-inflammatory. ABSTRACT: Titanium (Ti) is currently the most common biomaterial used for orthopedic implants; however, these implants may cause deleterious immune response. To investigate the possible mechanisms involved in excessive inflammation, we assessed the activity of neutrophils and monocyte-derived macrophages (MDMs) during the insertion of the Ti implant in a sheep model. The study was conducted on 12 sheep, 4 of which were control animals and 8 were in the experimental group with inserted Ti implant. Neutrophil secretory response was estimated at two time points T0 before surgery and T1 1 h after implantation and was based on the release of enzymes from neutrophil granules and reactive oxygen and nitrogen species (RONS) generation. MDM function was evaluated 5 months after implantation, on the basis of RONS generation arginase activity and morphological changes. Moreover, the influence of some autologous neutrophil derived products, namely, antimicrobial neutrophil extract (ANE) and neutrophil degranulation products (DGP) on leukocytes was estimated. Our study revealed that Ti implant insertion did not cause any adverse effects up to 5 months after surgical procedure. Stimulation of neutrophil cultures with ANE decreased the enzyme release as well as superoxide generation. Treatment of MDM with ANE diminished superoxide and NO generation and increased arginase activity. On the other hand, MDM stimulated with DGP showed elevated superoxide and NO generation as well as decreased arginase activity. To summarize, ANE exerted an anti-inflammatory and pro-resolving effect on studied leukocytes, whereas DGP acted as pro-inflammatory.