Effects of allogeneic and xenogeneic biomaterials on macrophage morphology and function in vitro and in vivo

Background. Determining the biocompatibility of biotransplants remains a pressing issue in tissue engineering. The degradation products of these materials elicit diverse cellular responses that ultimately influence healing outcomes. Among these responses, macrophages play a central role in regulating biocompatibility and guiding the regenerative process. Objective: to characterize the cellular composition and repertoire of macrophages induced by exogenous allogeneic and xenogeneic matrices, and to determine the morphofunctional profile of macrophages in vitro and in vivo following allogeneic and xenogeneic transplantation. Materials and methods. In vitro, donor-derived monocytes were cultured for 7 days on a substrate composed of a sponge matrix and a suspension of dispersed allogeneic biomaterials from the Alloplant® series, obtained from human cadaver tissue. In vivo, male outbred rats (200–250 g; n = 20) received subcutaneous injections of a suspension (10 mg) of dispersed allogeneic biomaterial (DAB) derived from rat tendons, and dispersed xenogeneic biomaterial (DXB) derived from rabbit tendons. Transplantation sites were excised at 7 and 14 days post-implantation. Morphological evaluation included histological and immunohistochemical analyses (VEGF-R1, CD206, CD86, TNF-α, CD163, TGF-β, CD68, FGF-1, MMP-9, TIMP-2, HLA-DR), as well as scanning electron microscopy. Results. In vitro, monocytes differentiated into mature macrophages exhibiting the following phenotype: CD68⁺, CD206⁺, VEGF-R⁺, CD86^–, TGF-β^–, CD163^–, FGF-1^–, MMP-9^–, and TIMP-2^–. In vivo, DAB underwent lysis with the release of glycosaminoglycans (GAG), followed by resorption and replacement without evidence of encapsulation by macrophages of the following phenotypes: M1 (CD86⁺) and M2 (CD206⁺, CD163⁺), as well as VEGF-R⁺ and TNF-α⁺ expression, while remaining negative for TGF-β and HLA-DR. In contrast, DXB induced encapsulation and granulomatous inflammation, with no detectable GAG release. Macrophages in this group predominantly exhibited an M2 phenotype (CD206⁺), with positive expression of TGF-β and HLA-DR, and negative expression of TNF-α and VEGF-R. Conclusion. The species specificity of biomaterials determines macrophage phenotype, cellular response, and healing outcomes. Allogeneic biomaterial was effectively resorbed and replaced by structurally complete regenerative tissue. Macrophages of both M1 and M2 phenotypes did not demonstrate antigenic or fibrogenic activity and promoted angiogenesis. In contrast, xenogeneic biomaterial elicited chronic inflammation and encapsulation, characterized by the presence of M2 macrophages with antigenic and fibrogenic properties.

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