Inhibition of Ferroptosis by Adipose Stem Cell-Derived Apoptotic Vesicles Enhances Angiogenesis and Accelerates Diabetic Wound Healing

Abstract

Jingyi Zhang,1,2,* Jinxin Kuang,1,* Shengkai Gong,1,3,* Hanzhe Wang,1 Feng Ding,1,3 Lu Zhao,1,4 Lele Shi,1,4 Shiyu Liu,1,3 Yimin Zhao,1 Jiani Liu,1,5 Geng Dou1,3 1State Key Laboratory of Oral & Maxillofacial Reconstruction and Regeneration, National Clinical Research Center for Oral Diseases, School of Stomatology, The Fourth Military Medical University, Xi’an, Shaanxi, 710032, People’s Republic of China; 2School of Basic Medicine, The Fourth Military Medical University, Xi’an, Shaanxi, 710032, People’s Republic of China; 3State Key Laboratory of Oral & Maxillofacial Reconstruction and Regeneration, National Clinical Research Center for Oral Diseases, Shaanxi Key Laboratory of Stomatology, Department of Oral Biology & Clinic of Oral Rare Diseases and Genetic Disease, School of Stomatology, the Fourth Military Medical University, Xi’an, Shaanxi, 710032, People’s Republic of China; 4State Key Laboratory of Oral & Maxillofacial Reconstruction and Regeneration, National Clinical Research Center for Oral Diseases, Shaanxi Clinical Research Center for Oral Diseases, Department of Oral and Maxillofacial Surgery, School of Stomatology, The Fourth Military Medical University, Xi’an, Shaanxi, 710032, People’s Republic of China; 5State Key Laboratory of Oral & Maxillofacial Reconstruction and Regeneration, National Clinical Research Center for Oral Diseases, Shaanxi Key Laboratory of Stomatology, Digital Center, School of Stomatology, The Fourth Military Medical University, Xi’an, Shaanxi, 710032, People’s Republic of China*These authors contributed equally to this workCorrespondence: Geng Dou, State Key Laboratory of Oral & Maxillofacial Reconstruction and Regeneration, National Clinical Research Center for Oral Diseases, School of Stomatology, The Fourth Military Medical University, No. 145, West Changle Road, Xi’an, Province Shaanxi, 710032, People’s Republic of China, Email [email protected] Jiani Liu, State Key Laboratory of Oral & Maxillofacial Reconstruction and Regeneration, National Clinical Research Center for Oral Diseases, Shaanxi Key Laboratory of Stomatology, Digital Center, School of Stomatology, The Fourth Military Medical University, No. 145, West Changle Road, Xi’an, Province Shaanxi, 710032, People’s Republic of China, Email [email protected]: Impaired angiogenesis is a critical challenge in diabetic wound healing. While apoptotic derivatives of stem cells hold promise for regenerative therapy, their role in modulating angiogenesis within the diabetic wound microenvironment remains underexplored. This study aims to investigate whether adipose stem cell-derived apoptotic vesicles (ASCs-apoVs) promote angiogenesis and accelerate diabetic wound healing by inhibiting endothelial cell ferroptosis.Methods: Diabetic mice model was established by feeding with high-fat diet (HFD) for 3 months followed by full-thickness skin wound preparation. Adipose stem cells (ASCs) isolated from adipose tissue were treated with staurosporine (STS) to induce apoptosis in vitro. Apoptotic vesicles (apoVs) were isolated by differential centrifugation, characterized using TEM, dynamic light scattering (DLS), and Western blot, and applied topically to diabetic wounds. The therapeutic effects of apoVs on wound healing efficiency, vascularization level and endothelial cell ferroptosis were evaluated.Results: HFD-induced diabetes promoted lipid peroxidation (4HNE accumulation) and ferroptosis in endothelial cells (ECs), leading to reduced CD31+ and vWf+ vessel density and delayed wound closure. In vitro diabetic endothelial cell models confirmed increased lipid peroxidation and ferroptosis, which compromised the proliferation, migration and tube formation capacities of ECs. ASCs-apoVs, characterized by typical extracellular vesicle (EV) morphology and apoptotic markers, significantly inhibited lipid peroxidation and ferroptosis of ECs, thereby promoting angiogenesis and accelerating diabetic wound healing.Conclusion: Ferroptosis of endothelial cells contributes to impaired vascularization in diabetic wounds. ApoVs represent a promising cell-free therapeutic approach to mitigate ferroptosis, restore endothelial function and promote angiogenesis, offering a potential strategy for diabetic wound management.Keywords: apoptotic vesicles, wound healing, vascularization, adipose stem cells, ferroptosis, endothelial cells

Keywords

• apoptotic vesicles
• wound healing
• vascularization
• adipose stem cells
• ferroptosis
• endothelial cells.