Polymeric Three-dimensional Scaffold Promotes Osteogenic Precursor Cells Differentiation through Induction of Peripheral Macrophages Polarization

doi:10.13701/j.cnki.kqyxyj.2024.03.004

Abstract

Abstract: Objective: To investigate the effect of three-dimensional scaffold SF-CS-HA on the polarization of mouse macrophages, and explore the impact of bone immunomodulatory microenvironment regulation on the extracortical bone formation of mouse calvarial precursor cells. Methods: SF-CS-HA was prepared and co-cultured with mouse macrophages RAW264.7. Flow cytometry was performed to detect the polarization of macrophages into M2 phenotype. RT-qPCR was used to measure the expression of inflammatory cytokine genes, and western blot was conducted to evaluate the expression of TGF-β. After co-incubation of macrophages with SF-CS-HA 3D, conditioned medium was prepared. The proliferation, migration, ALP activity, and morphological changes of mouse calvarial precursor cells MC3T3-E1 were compared between complete medium and conditioned medium. Western blot was performed to detect the expression of bone formation-related proteins. An animal model was used to verify the effect of SF-CS-HA on the increase of skull bone mass in mice. Results: Scanning electron microscopy observations revealed that SF-CS-HA had a uniform porous structure both on the surface and internally. Flow cytometry analysis showed that SF-CS-HA promoted the polarization of peripheral macrophages towards M2 phenotype. RT-qPCR and western blot results demonstrated that SF-CS-HA increased the expression of IL-10 gene and TGF-β protein, while decreased the expression of IL-1β gene. Additionally, SF-CS-HA promoted the proliferation of MC3T3-E1 cells, significantly increased ALP activity, enhanced cell migration, and increased the production of cytoskeletal protein Factin. Furthermore, conditioned medium exhibited more significant enhancement in the proliferation and migration capabilities of MC3T3-E1 cells. Moreover, Western blot indicated that under the regulation of the bone immune microenvironment, the expression of bone formation proteins OPN, COLA1, RUNX2, OCN, and TGF-β increased significantly. The results of animal experiments showed that SF-CS-HA could promote new bone formation in the mouse skull in vivo. Conclusion: SF-CS-HA 3D scaffold can promote the polarization of peripheral macrophages and regulate the bone immune microenvironment, thereby promoting osteogenic differentiation and bone formation in the cortical bone exogenesis.

Key words: 3D scaffold material, macrophages, polarization, co-culture, calvarial precursor cells

LIU Yu, HUANG Weikun, LIANG Yu. Polymeric Three-dimensional Scaffold Promotes Osteogenic Precursor Cells Differentiation through Induction of Peripheral Macrophages Polarization[J]. Journal of Oral Science Research, 2024, 40(3): 206-213.

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