Biomaterials with mechanical or biological competence are ubiquitous in musculoskeletal disorders, and understanding the inflammatory response they trigger is key to guide tissue regeneration. While macrophage role has been widely investigated, immune response is regulated by other immune cells, including neutrophils, the most abundant leukocyte in human blood. As first responders to injury, infection or material implantation, neutrophils recruit other immune cells, and therefore influence the onset and resolution of chronic inflammation, and macrophage polarization. This response depends on the physical and chemical properties of the biomaterials, among other factors. In this study we report an in vitro culture model to describe the most important neutrophil functions in relation to tissue repair. We identified neutrophil survival and death, neutrophils extracellular trap formation, release of reactive oxygen species and degranulation with cytokines release as key functions and introduced a corresponding array of assays. These tests were suitable to identify clear differences in the response by neutrophils that were cultured on material of different origin, stiffness and chemical composition. Overall, substrates from biopolymers of natural origin resulted in increased survival, less neutrophil extracellular trap formation, and more reactive oxygen species production than synthetic polymers. Within the range of mechanical properties explored (storage modulus below 5 k Pa), storage modulus of covalently crosslinked hyaluronic acid hydrogels did not significantly alter neutrophils response, whereas polyvinyl alcohol gels of matching mechanical properties displayed a response indicating increased activation. Additionally, we present the effect of material stiffness, charge, coating and culture conditions in the measured neutrophils response. Further studies are needed to correlate the neutrophil response to tissue healing. By deciphering how neutrophils initiate and modulate the immune response to material implantation, we aim at introducing new principles to design immunomodulatory biomaterials for musculoskeletal disorders.
This work was supported by the AO Foundation, AO CMF, grant AOCMF-21-04S.
A study was undertaken to assess the degree of inter-observer error when a panel of observers classified the radiographs of patients with early Perthes' disease, using Catterall grouping and "at risk" signs. The anteroposterior and lateral radiographs, taken within three months of diagnosis of Perthes' disease, were available for 69 hips and were shown in turn to 10 observers. The radiological end-results were assessed at least four years from diagnosis. The results showed a poor ability of the observers to delineate Groups 1, 2 and 3, with a more satisfactory performance in Group 4 and when Groups 2 and 3 were combined. Interpretation of "at risk" signs was unsatisfactory except when there was an increase in medial joint space greater than two millimetres. The end-results correlated well with early Catterall grouping and "at risk" signs when these were correctly interpreted.
