ADVANCED m-journal de CFO O-E/M CFO 10-E/M CFO O-E CFO 10-E e CFO 10-E/M CFO 10-E CFO O-E O O-M CFO 10-E/M 10-E/M VSI Pathway Enrichm d Figure 5. CFO/P(VDF-TrFE)magnetoelectric nanocomposite membrane mediated bone regeneration and immune response in vivo. a)Representa- tive CT images of bone regeneration within rat cranial defects at 8 weeks after membrane implantation, with the CFo 10-E/M group exhibiting the st abundant new bone formation. Yellow arrows indicate enhanced bone regeneration in the CFo 10-E/M group. Yellow triangles denote new bone denote the boundary between nascent bone and host bone. b)Quantitative analysis of the total volume of newly formed bone ted in the ceo 1o-E/M group. Blue arrow indicates the enriched genes in the cop at some immune-related proteins and pathways were ssue.(* VS CFO 10-E/M, p<0.05)c)KEGG pathway analysis, d )heat map demonstrating t ment and coagulation cascades pathway of the CFo 10-E/M group. e)Immunofluorescence images of CD206 expression, indicating that the CFo 10-E/M membrane promoted adherent macrophage M2 polarization(Scale bars: 50 um) bone regeneration. After initial inflammation, the phenotype 3. Conclusion was transitioned to M2 at the remodeling stage. M2 macro- phages are known as the pro- healing phenotype. 46 Many This study developed a flexible and reusable magnetoelec biomaterials have been designed to promote tissue regenera- tric nanocomposite membrane for bone regeneration which tion by activating M2 polarization of macrophages, I/Taken can be regulated by a remote dC magnetic field to mimic the together, these results thus indicated that the CFO 10.E/M natural magnetoelectric microenvironment. Based on MD membranes could activate the initial immune response and simulations together with biological evaluation, the 10 wt% accelerate the transition from M1 to M2 phenotype to further CFO/P(VDF-TrFE) magnetoelectric nanocomposite mem- promote bone regeneration. Therefore, our findings sug. branes were confirmed to be the optimal group for promoting gested that the CFo 10-E/M membrane could provide a mag. bone regeneration by increasing RGD exposure. Moreover, the netoelectric microenvironment with 12h remote DC magnetic magnetoelectric microenvironment provided by the magneto- field shifting to enhance bone regeneration by activating the electric nanocomposite membrane not only directly enhanced me response and accelerating the transition from the BM-MSC osteogenic differentiation, but also regulated the oste- Immune ammation stage to bone healing stage. iMmuno modulatory environment to improve bone regenera The remotely tuned magnetoelectric microenvironment pro- tion. The osteoimmunomodulatory microenvironment wit vided by the removable and easily shaping membrane have the the bone defect area triggered initial inflammation and then potential to be utilized clinically for space maintenance and subsequently promoted MI to M2 transition of macrophages Our research thus provides a novel strategy of remote tuning Adv Funct. Mater. 2020. 2006226 20062269of1) o 2020 Wiley-VCH GmbHwww.advancedsciencenews.com www.afm-journal.de 2006226 (9 of 11) © 2020 Wiley-VCH GmbH bone regeneration. After initial inflammation, the phenotype was transitioned to M2 at the remodeling stage. M2 macro￾phages are known as the pro-healing phenotype.[46] Many biomaterials have been designed to promote tissue regenera￾tion by activating M2 polarization of macrophages.[47] Taken together, these results thus indicated that the CFO 10-E/M membranes could activate the initial immune response and accelerate the transition from M1 to M2 phenotype to further promote bone regeneration. Therefore, our findings sug￾gested that the CFO 10-E/M membrane could provide a mag￾netoelectric microenvironment with 12h remote DC magnetic field shifting to enhance bone regeneration by activating the immune response and accelerating the transition from the acute inflammation stage to bone healing stage. The remotely tuned magnetoelectric microenvironment pro￾vided by the removable and easily shaping membrane have the potential to be utilized clinically for space maintenance and bone regeneration. 3. Conclusion This study developed a flexible and reusable magnetoelec￾tric nanocomposite membrane for bone regeneration which can be regulated by a remote DC magnetic field to mimic the natural magnetoelectric microenvironment. Based on MD simulations together with biological evaluation, the 10 wt% CFO/P(VDF-TrFE) magnetoelectric nanocomposite mem￾branes were confirmed to be the optimal group for promoting bone regeneration by increasing RGD exposure. Moreover, the magnetoelectric microenvironment provided by the magneto￾electric nanocomposite membrane not only directly enhanced BM-MSC osteogenic differentiation, but also regulated the oste￾oimmunomodulatory environment to improve bone regenera￾tion. The osteoimmunomodulatory microenvironment within the bone defect area triggered initial inflammation and then subsequently promoted M1 to M2 transition of macrophages. Our research thus provides a novel strategy of remote tuning Figure 5. CFO/P(VDF-TrFE) magnetoelectric nanocomposite membrane mediated bone regeneration and immune response in vivo. a) Representa￾tive CT images of bone regeneration within rat cranial defects at 8 weeks after membrane implantation, with the CFO 10-E/M group exhibiting the most abundant new bone formation. Yellow arrows indicate enhanced bone regeneration in the CFO 10-E/M group. Yellow triangles denote new bone. Yellow dotted lines denote the boundary between nascent bone and host bone. b) Quantitative analysis of the total volume of newly formed bone tissue. (* VS CFO 10-E/M, p < 0.05) c) KEGG pathway analysis, d) heat map demonstrating that some immune-related proteins and pathways were upregulated in the CFO 10-E/M group. Blue arrow indicates the enriched genes in the complement and coagulation cascades pathway of the CFO 10-E/M group. e) Immunofluorescence images of CD206 expression, indicating that the CFO 10-E/M membrane promoted adherent macrophage M2 polarization (Scale bars: 50 µm). Adv. Funct. Mater. 2020, 2006226