TY - JOUR
T1 - Crosstalk between macrophages and mesenchymal stem cells shape patterns of osteogenesis and immunomodulation in mineralized collagen scaffolds
AU - Kolliopoulos, Vasiliki
AU - Polanek, Maxwell
AU - Wong Yan Ling, Melisande
AU - Tiffany, Aleczandria
AU - Spiller, Kara L.
AU - Harley, Brendan A.C.
N1 - This work shows macrophage secretome has the potential to educate MSCs towards a more immunomodulatory phenotype as well as the potential to enhance MSC osteogenesis regardless of initial licensing state. We used separated culture for its ability to resolve the paracrine effects of macrophages on MSCs in a bone mimicking environment. Basal MSCs displayed significantly greater OPG release (Fig. 2A), while licensed MSCs display significantly greater osteogenic gene expression (Fig. 3B), in the presence of M1 macrophage secretome compared to all other groups. Vasandan et al., reported that MSCs in the presence of M1 (IFN-\u03B3/LPS) macrophages in 2D culture, decrease their pro-inflammatory secreted markers and polarize them towards an M2-like phenotype [57]. Our data would support this argument as an increase in OPG release would demonstrate an enhanced osteoblastic activity indicative of the remodeling phase post inflammation. Licensed MSCs display significantly higher expression of PGE2 on Day 7 of culture in the presence of M0 macrophage secretome than all other groups (Fig. 2B), and significantly higher mid-to-late-culture expression of immunomodulatory genes such as IL1RN and IL-6 in the presence of M1 genes (Fig. 3C). Interestingly, when compared to our previous single culture study of MSCs on mineralized collagen scaffold in basal and licensed conditions [36], we observe that the presence of macrophage secretome upregulates MSC osteogenesis via enhanced OPG expression in all groups and decreases MSC immunomodulatory activity in licensed groups via IL-6 and PGE2 production [36]. However, MSC licensing had a greater effect on immunomodulatory gene expression, with a majority of assayed genes significantly (p < 0.05) increased on Day 1 in response to MSC licensing.The authors would like to acknowledge the following institutes for access to their facilities and services: the School of Chemical Sciences Microanalysis Laboratory, the Carl R. Woese Institute for Genomic Biology, Dr. Hui Xu, and the Tumor Engineering and Phenotyping Shared Resource (TEP) at the Cancer Center at Illinois, and the Beckman Institute for Advanced Science and Technology, all located at the University of Illinois. Research reported in this publication was supported by the National Institute of Dental and Craniofacial Research of the National Institutes of Health under Award Number R21 DE026582and R01 DE030491(BACH), as well as the National Institute of Arthritis and Musculoskeletal and Skin Diseases under Award Number R01 AR077858(BACH). We are also grateful for funds provided by the NSF Graduate Research Fellowship (DGE-174604to VK; DGE-1144245 to AST) and the Chemistry-Biology Interface Research Training Program at the University of Illinois (T32 GM070421, VK). Additional support was provided by the Carl R. Woese Institute for Genomic Biology and the Chemical and Biomolecular Engineering Dept. at the University of Illinois at Urbana-Champaign. The interpretations and conclusions presented are those of the authors and are not necessarily endorsed by the National Institutes of Health or the National Science Foundation.
The authors would like to acknowledge the following institutes for access to their facilities and services: the School of Chemical Sciences Microanalysis Laboratory, the Carl R. Woese Institute for Genomic Biology, Dr. Hui Xu, and the Tumor Engineering and Phenotyping Shared Resource (TEP) at the Cancer Center at Illinois, and the Beckman Institute for Advanced Science and Technology, all located at the University of Illinois. Research reported in this publication was supported by the National Institute of Dental and Craniofacial Research of the National Institutes of Health under Award Number R21 DE026582 and R01 DE030491 (BACH), as well as the National Institute of Arthritis and Musculoskeletal and Skin Diseases under Award Number R01 AR077858 (BACH). We are also grateful for funds provided by the NSF Graduate Research Fellowship (DGE-174604 to VK; DGE-1144245 to AST) and the Chemistry-Biology Interface Research Training Program at the University of Illinois (T32 GM070421, VK). Additional support was provided by the Carl R. Woese Institute for Genomic Biology and the Chemical and Biomolecular Engineering Dept. at the University of Illinois at Urbana-Champaign. The interpretations and conclusions presented are those of the authors and are not necessarily endorsed by the National Institutes of Health or the National Science Foundation.
PY - 2025/2
Y1 - 2025/2
N2 - 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 in vitro 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.
AB - 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 in vitro 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.
KW - Co-cultures
KW - Immunomodulation
KW - Macrophages
KW - Mesenchymal stem cells
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U2 - 10.1016/j.bioactmat.2024.09.030
DO - 10.1016/j.bioactmat.2024.09.030
M3 - Article
AN - SCOPUS:85205787317
SN - 2452-199X
VL - 44
SP - 34
EP - 45
JO - Bioactive Materials
JF - Bioactive Materials
ER -