Abstract
Purpose To study the effect of osteoplastic materials based on the extracellular xenomatrix of bovine and equine bone tissue on the formation of neutrophil extracellular traps (NETs) in the peripheral blood of rabbits in the early post-operative period after implantation. Materials and methods The study was carried out on 18 male rabbits of the Soviet Chinchilla breed, aged from 8 months to 1.2 years, weighing from 3.0 to 4.5 kg. A perforated bone defect of a cylindrical shape measuring 2 x 6 mm in the distal metaphysis of the right and left femurs was modeled in the animals. The rabbits were divided into three groups, six animals each. In group I, the bone defect was left unfilled; in group II, the defect was filled with a bovine bone tissue xenomatrix, and an equine bone tissue xenomatrix was implanted in group III animals. The implantation material had the appearance of a yellowish crumb with a particle size of 0.5– 1 mm. Blood smears stained according to Romanovsky-Giemsa were used for counting extracellular neutrophil traps (NETs). The percentage of neutrophils that passed the stages of nuclear transformation and emitted free chromatin into the extracellular space in the form of network-like structures was calculated. Results On days 3–7 of the experiment, the number of NETs increased in the early stages of NETosis in all groups. There were no significant differences between the groups. In group I, on days 7 and 14, the number of early forms of NETs (stages 1a and 1b) returned to the values of the preoperative period. In groups II and III, normalization of NETs (stage 1a) did not occur, and the content of NETs (stage 1b) returned to the initial level only by day 30 of the experiment. On days 3, 7, 14, the number of mature NETs increased in all groups. The highest values were noted in group II, where the bovine xenogeneic matrix was implanted. Conclusion Implantation materials based on the extracellular matrix of equine and bovine xenogeneic bone stimulate excessive formation of early NETs on days 14–30 of the experimental period in response to xenotransplantation. Xenomaterials of bovine bone tissue, in comparison with xenomaterials of equine bone tissue, induce a more pronounced inflammatory reaction in the nearest time after defect filling, which is manifested by higher production of mature NETs on days 3–14 of the experiment.
Highlights
Bone defects resulting from injuries, diseases, surgical interventions or congenital disorders remain challenging problems in contemporary orthopaedics and traumatology [1].There is an extremely high demand for bone grafts currently
Analysis of literature sources showed that the following immunological parameters and tests were used to assess the immune response and inflammation activity in xenoimplantation: TNFа, TNFβ, IFN-γ, IL-2, IL-4, IL-6, IL-10, TGF, IL- 12P70, IL-12, immunoglobulins, lymphocyte subpopulations, C-reactive protein, lymphocyte transformation test (LTT), memory lymphocyte immunostimulation assay (MELISA), leukocyte migration inhibition test (LIF), lymphocyte activation test (LAT) [11-15]
The probable reason may be the fact that the third protective strategy of neutrophils was discovered quite recently (2004), and the study of NETosis processes in various pathological conditions and surgical interventions has been at the stage of data accumulation [16]
Summary
Bone defects resulting from injuries, diseases, surgical interventions or congenital disorders remain challenging problems in contemporary orthopaedics and traumatology [1].There is an extremely high demand for bone grafts currently. An alternative to autoplasty is the use of xenomaterials from bone tissue, which, in turn, contain foreign antigens in their structure. It can cause biological incompatibility, leading to the development of an immune cell-mediated rejection, an acute humoral response, initially mediated by the formation of IgM antibodies, mainly to the gal epitope, with a subsequent increase in the level of IgG, overproduction of cytokines [3, 4]. The role of inflammation has not been fully understood if bone tissue xenomatrix is applied. The study of immunological parameters may be used to assess the course of the postoperative period after implantation xenomaterials from bone tissue and the "survival" of the implant
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