Small extracellular vesicles from deferoxamine-primed stem cells promote angiogenesis and tissue integration

Abstract

Aim: We investigated whether the secretome, particularly small extracellular vesicles (sEVs), obtained from deferoxamine (DFO)-pretreated stem cells from human exfoliated deciduous teeth (SHED), could promote angiogenesis in human umbilical vein endothelial cells (HUVECs) and support vascularized tissue integration under preliminary ectopic implantation conditions.

Methods: SHED were treated with DFO (2.5, 5, 10 μM), and RNA sequencing was performed to analyze gene expression changes in DFO-treated SHED. The effects of the DFO-pretreated SHED secretome and the isolated sEVs (DFO-sEVs) on HUVEC migration were evaluated. The angiogenic capacity was assessed using wound healing, Transwell migration, and tube formation assays. DFO-sEVs were also tested for their ability to regulate macrophage polarization using immunofluorescence staining. Finally, DFO-sEVs were encapsulated in hydrogels and subcutaneously implanted into Sprague-Dawley rats, and their pro-angiogenic capacity in vivo was assessed via histological staining.

Results: RNA-seq analysis indicated that DFO pretreatment modulated SHED pathways related to hypoxia response, cell adhesion, and inflammation. The secretome from SHED pretreated with 5 μM DFO significantly promoted HUVEC migration and tube formation. DFO-sEVs, particularly from the 5 μM DFO group, demonstrated superior pro-angiogenic effects compared to sEVs from untreated SHED both in vitro and in vivo. Furthermore, DFO-sEVs drove macrophages to adopt an M2-like reparative polarization.

Conclusion: DFO pretreatment functionally potentiates the pro-angiogenic and immunomodulatory properties of SHED-derived sEVs. The findings suggest that DFO-sEVs may provide a bioactive component for promoting angiogenesis and facilitating tissue integration.

Keywords

Angiogenesis / deferoxamine / stem cells from human exfoliated deciduous teeth / small extracellular vesicles / hydrogel

References

[1] Chang C,Yan J,Yao Z,Zhang C,Li X,Mao HQ. Effects of mesenchymal stem cell-derived paracrine signals and their delivery strategies Adv Healthc Mater 2021 10 e2001689 PMC7995150

[2] Trigo CM,Rodrigues JS,Camões SP,Solá S,Miranda JP. Mesenchymal stem cell secretome for regenerative medicine: where do we stand? J Adv Res 2025;70:103-24 PMC11976416

[3] Mahdavi-Jouibari F,Parseh B,Kazeminejad E,Khosravi A. Hopes and opportunities of stem cells from human exfoliated deciduous teeth (SHED) in cartilage tissue regeneration Front Bioeng Biotechnol 2023 11 1021024 PMC9968979

[4] Kunimatsu R,Hiraki T,Rikitake K.et al. Effects of human deciduous dental pulp-derived mesenchymal stem cell-derived conditioned medium on the metabolism of HUVECs, osteoblasts, and BMSCs Cells 2022 11 3222 PMC9600042

[5] Rogulska O,Vackova I,Prazak S.et al. Storage conditions affect the composition of the lyophilized secretome of multipotent mesenchymal stromal cells Sci Rep 2024 14 10243 PMC11068735

[6] Chouaib B,Haack-Sørensen M,Chaubron F,Cuisinier F,Collart-Dutilleul PY. Towards the standardization of mesenchymal stem cell secretome-derived product manufacturing for tissue regeneration Int J Mol Sci 2023 24 12594 PMC10454619

[7] Gao Y,Sang J,Wan Z.et al. Advanced small extracellular vesicles delivery systems for in situ tissue engineering Extracell Vesicles Circ Nucl Acids 2026 7 354 76 PMC13074283

[8] Welsh JA,Goberdhan DCI,O'Driscoll L.et al. ; MISEV Consortium. Minimal information for studies of extracellular vesicles (MISEV2023): from basic to advanced approaches J Extracell Vesicles 2024 13 e12404 PMC11082720

[9] Wu J,Chen L,Wang R.et al. Exosomes secreted by stem cells from human exfoliated deciduous teeth promote alveolar bone defect repair through the regulation of angiogenesis and osteogenesis ACS Biomater Sci Eng 2019 5 3561 71

[10] Gao Y,Yuan Z,Yuan X.et al. Bioinspired porous microspheres for sustained hypoxic exosomes release and vascularized bone regeneration Bioact Mater 2022 14 377 88 PMC8964815

[11] Xie Y,Yu L,Cheng Z.et al. SHED-derived exosomes promote LPS-induced wound healing with less itching by stimulating macrophage autophagy J Nanobiotechnology 2022 20 239 PMC9124392

[12] Jin S,Wang Y,Wu X.et al. Young exosome bio-nanoparticles restore aging-impaired tendon stem/progenitor cell function and reparative capacity Adv Mater 2023 35 e2211602

[13] Long R,Wang S. Exosomes from preconditioned mesenchymal stem cells: tissue repair and regeneration Regen Ther 2024 25 355 66 PMC10875222

[14] Ma M. Role of hypoxia in mesenchymal stem cells from dental pulp: influence, mechanism and application Cell Biochem Biophys 2024 82 535 47 PMC11344735

[15] Siavashi V,Nassiri SM,Farhadi Mahalli M,Kamarul T,Sharifi AM. Elucidating the effect of deferoxamine, a hypoxia mimetic agent, on angiogenesis restoration in endothelial progenitor cells (EPCs) from diabetic mice Iran J Basic Med Sci 2025 28 1589 97 PMC12571184

[16] Song YC,Park GT,Moon HJ.et al. Hybrid spheroids containing mesenchymal stem cells promote therapeutic angiogenesis by increasing engraftment of co-transplanted endothelial colony-forming cells in vivo Stem Cell Res Ther 2023 14 193 PMC10394850

[17] Liu P,Qin L,Liu C.et al. Exosomes derived from hypoxia-conditioned stem cells of human deciduous exfoliated teeth enhance angiogenesis via the transfer of let-7f-5p and miR-210-3p Front Cell Dev Biol 2022 10 879877 PMC9086315

[18] Zhu Y,Liao ZF,Mo MH,Xiong XD. Mesenchymal stromal cell-derived extracellular vesicles for vasculopathies and angiogenesis: therapeutic applications and optimization Biomolecules 2023 13 1109 PMC10377109

[19] Du W,Zhang K,Zhang S.et al. Enhanced proangiogenic potential of mesenchymal stem cell-derived exosomes stimulated by a nitric oxide releasing polymer Biomaterials 2017 133 70 81

[20] Huang D,Shen H,Xie F.et al. Role of mesenchymal stem cell-derived exosomes in the regeneration of different tissues J Biol Eng 2024 18 36 PMC11155050

[21] Ratajczak J,Miekus K,Kucia M.et al. Embryonic stem cell-derived microvesicles reprogram hematopoietic progenitors: evidence for horizontal transfer of mRNA and protein delivery Leukemia 2006 20 847 56

[22] Valadi H,Ekström K,Bossios A,Sjöstrand M,Lee JJ,Lötvall JO. Exosome-mediated transfer of mRNAs and microRNAs is a novel mechanism of genetic exchange between cells Nat Cell Biol 2007 9 654 9

[23] Huang K,Mi B,Xiong Y.et al. Angiogenesis during diabetic wound repair: from mechanism to therapy opportunity Burns Trauma 2025 13 tkae052 PMC11802347

[24] Yi C,Wu W,Zheng D.et al. Targeted inhibition of endothelial calpain delays wound healing by reducing inflammation and angiogenesis Cell Death Dis 2020 11 533 PMC7360547

[25] Galiano RD,Tepper OM,Pelo CR.et al. Topical vascular endothelial growth factor accelerates diabetic wound healing through increased angiogenesis and by mobilizing and recruiting bone marrow-derived cells Am J Pathol 2004 164 1935 47 PMC1615774

[26] Sharifiaghdam M,Shaabani E,Faridi-Majidi R,De Smedt SC,Braeckmans K,Fraire JC. Macrophages as a therapeutic target to promote diabetic wound healing Mol Ther 2022 30 2891 908 PMC9482022

[27] Peña OA,Martin P. Cellular and molecular mechanisms of skin wound healing Nat Rev Mol Cell Biol 2024 25 599 616

[28] Wetzler C,Kämpfer H,Stallmeyer B,Pfeilschifter J,Frank S. Large and sustained induction of chemokines during impaired wound healing in the genetically diabetic mouse: prolonged persistence of neutrophils and macrophages during the late phase of repair J Invest Dermatol 2000 115 245 53

[29] Bannon P,Wood S,Restivo T,Campbell L,Hardman MJ,Mace KA. Diabetes induces stable intrinsic changes to myeloid cells that contribute to chronic inflammation during wound healing in mice Dis Model Mech 2013 6 1434 47 PMC3820266