Viscoelastic Hydrogel Modulates Phenotype of Macrophage‐Derived Multinucleated Cells and Macrophage Differentiation in Foreign Body Reactions

ABSTRACT Biomaterial‐induced macrophage‐derived multinucleated cells (MNCs) are often observed at or near material implantation sites, yet their subtypes and roles in tissue repair and wound healing remain unclear. This study compares material‐induced MNCs to cytokine‐induced MNCs using both in vitro and in vivo models. 3D‐embedded Raw264.7 cells and rat bone marrow‐derived monocytes (BMDMs), with or without cytokines such as IL‐4 and RANKL, were characterized for their MNC morphologies and subtypes via in situ immunocytochemistry and flow cytometry. Macrophage polarization and osteoclastic differentiation were assessed through NO production, arginase activity, and tartrate‐resistant acid phosphatase levels. 3D matrix‐induced MNCs expressed the same phenotypic heterogeneity as the IL‐4 and RANK‐treated ones. 3D matrix‐induced MNCs displayed the same phenotypic heterogeneity as those treated with IL‐4 and RANKL. A high viscoelastic matrix (1006.48 ± 92.29 Pa) induced larger populations of proinflammatory and osteoclast‐like MNCs, whereas a low viscoelastic matrix (38.61 ± 7.56 Pa) supported active differentiation and gene expression across pro‐, anti‐inflammatory, and osteoclast‐like macrophages. Matrix viscoelasticity also influenced the effects of IL‐4 and RANKL on macrophage‐derived MNC polarization. In an in vivo subcutaneous implantation model, medium to high viscoelastic matrices exhibited higher populations of CD86+ and RANK+ MNCs, while low viscoelastic matrices showed higher populations of CD206+ MNCs. These findings suggest that matrix viscoelasticity modulates macrophage differentiation and MNC phenotype, with low viscoelastic matrices potentially favoring anti‐inflammatory MNCs and macrophage differentiation suitable for subcutaneous implantation.