Mechanism by which magnesium implant-activated integrin α10β1 promotes osteogenic differentiation of periosteal stem cells

doi:10.12307/2026.456

Abstract

Abstract: BACKGROUND: Periosteal stem cells are the key cellular population in magnesium-induced osteogenesis. Integrin α10β1 is a magnesium-dependent heterodimeric adhesion molecule. Mg²⁺ binds to the MIDAS domain of integrin α10β1, functioning as a molecular switch to regulate downstream biological processes.
OBJECTIVE: To elucidate the mechanism by which magnesium-based implants regulate osteogenic differentiation of periosteal stem cells by activating integrin α10β1.
METHODS: Forty-two C57BL/6 mice were randomly divided into a titanium rod implantation group (n=21) and a magnesium rod implantation group (n=21). Titanium rods and magnesium rods were implanted into the medullary cavity of the intercondylar fossa of the left knee joint femur, respectively. Three days post-surgery, samples were collected for TUNEL staining to observe cell apoptosis around the implants. EdU staining was utilized to observe cell proliferation activity in the cortical bone thickening area. Fourteen days post-surgery, samples were collected for Micro-CT analysis of cortical bone thickening and osteogenesis. Hematoxylin-eosin staining was applied to observe the morphology and structure of newly formed bone in the cortical bone thickening area. Calcein green fluorescent double staining was employed to analyze the osteogenic differentiation of periosteal stem cells. qPCR was used to detect the expression of osteogenic marker genes Runx2, osteoblast-specific transcription factor, alkaline phosphatase, osteopontin, and integrin α10 and integrin β1. Western blot assay was utilized to detect the expression of integrin α10, integrin β1, focal adhesion kinase and phosphorylated focal adhesion kinase proteins. Transcriptome sequencing was applied to analyze the expression of Wnt/β-catenin and mitogen-activated protein kinase signaling pathways and the correlation between integrin α10β1 and osteogenic gene expression.
RESULTS AND CONCLUSION: (1) TUNEL staining and EdU staining showed a significant accumulation of apoptotic cells around the titanium rods, with only a few proliferating cells observed in the periosteum and no osteogenic differentiation. In contrast, no apoptotic cells were found around the magnesium rods, and a marked increase in proliferating cells was observed in the thickened periosteal region. (2) Micro-CT analysis revealed that the titanium rods did not degrade, while the magnesium rods showed significant degradation. The magnesium rod implantation group exhibited superior cortical bone thickening and osteogenesis compared with the titanium rod implantation group. Hematoxylin-eosin staining and calcein green fluorescence double staining further confirmed that osteogenesis was more pronounced in the magnesium rod implantation group. qPCR results indicated that the mRNA expression of Runx2, osteoblast-specific transcription factors, alkaline phosphatase, osteopontin, integrin α10, and integrin β1 was higher in the magnesium rod implantation group compared with the titanium rod implantation group. Western blot results showed higher expression of integrin α10, integrin β1, focal adhesion kinase (FAK), phosphorylated focal adhesion kinase, and Wnt/β-catenin in the magnesium rod implantation group, whereas mitogen-activated protein kinase (MAPK) expression was lower than in the titanium rod implantation group. Transcriptome sequencing analysis revealed a significant positive correlation between integrin α10β1 and the expression of Runx2, osteoblast-specific transcription factors, alkaline phosphatase, and osteopontin. (3) Conclusion: The results suggest that magnesium-based implants promote osteogenic differentiation of periosteal stem cells by activating the integrin α10β1-focal adhesion kinase/phosphorylated focal adhesion kinase signaling pathway. This activation upregulates Wnt/β-catenin signaling and inhibits certain MAPK signaling pathways, thus facilitating osteogenesis.

Key words: magnesium implants, integrin α10β1, osteogenic differentiation, periosteal stem cells, bone regeneration, Wnt/β-catenin signaling pathway, bone repair, osteoporosis

CLC Number:

Yang Yanjun, Zhu Lin, Gu Yongchun, Yan Zhanjun. Mechanism by which magnesium implant-activated integrin α10β1 promotes osteogenic differentiation of periosteal stem cells[J]. Chinese Journal of Tissue Engineering Research, 2026, 30(32): 8319-8326.

Figures/Tables 5

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