Crosstalk between macrophages and mesenchymal stem cells shape patterns of osteogenesis and immunomodulation in mineralized collagen scaffolds

Mesenchymal stem cells (MSCs) are highly plastic, with the capacity to differentiate into a spectrum of tissue-specific stromal cells. In the field of bone regeneration, MSCs have largely been considered for their osteogenic differentiation capacity. MSCs are increasingly being appreciated for their immunomodulatory potential following exposure to pro-inflammatory stimuli (licensing). Pro-inflammatory environments arise following bone injury via activation of resident immune cells like macrophages. We describe the use of a mineralized collagen scaffold as a bone-mimetic <i>in vitro</i> model to study the influence of paracrine versus direct cell-to-cell contact of THP-1 macrophages on MSC osteogenic and immunomodulatory potential. Paracrine stimuli from macrophages enhance MSC osteogenic and immunomodulatory potential via upregulation of key transcriptomic markers as well as via soluble biomolecule production. Direct co-culture of MSCs and macrophages decreased immunomodulatory potential in MSCs, especially for licensed MSCs, but enhanced matrix remodeling and expression of genes related to macrophage chemotaxis. These data demonstrate the significant effect macrophage-derived paracrine factors and direct contact have on MSC activity in a biomaterial model of bone regeneration. This work illuminates a critical need to further understand these processes in more clinically relevant cell models to inform biomaterial design.