多余牙源性牙髓干细胞来源的凋亡细胞外囊泡通过 PI3K/Akt/VEGF 通路转移 COL1A1 促进血管生成。
Apoptotic Extracellular Vesicles from Supernumerary Tooth-Derived Pulp Stem Cells Transfer COL1A1 to Promote Angiogenesis via PI3K/Akt/VEGF Pathway.
机构信息
Department of Pediatric Dentistry, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, College of Stomatology, Shanghai Jiao Tong University, National Center for Stomatology, National Clinical Research Center for Oral Diseases, Shanghai Key Laboratory of Stomatology, Shanghai Research Institute of Stomatology, Shanghai, People's Republic of China.
Shanghai Engineering Research Center of Advanced Dental Technology and Materials, Shanghai, People's Republic of China.
出版信息
Int J Nanomedicine. 2024 Jul 8;19:6811-6828. doi: 10.2147/IJN.S466136. eCollection 2024.
PURPOSE
Angiogenesis is a tightly controlled process that initiates the formation of new vessels and its dysfunction can lead to life-threatening diseases. Apoptotic extracellular vesicles (ApoEVs) have emerged as a proangiogenic agent with high safety and isolation efficiency profile, and ApoEVs from supernumerary tooth-derived pulp stem cells (SNTSC-ApoEVs) have their unique advantages with an easily accessible parental cell source and non-invasive cell harvesting. However, the detailed characteristics of SNTSC-ApoEVs are largely unknown. This study aimed to investigate the proangiogenic capacity and function molecule of SNTSC-ApoEVs.
METHODS
SNTSC-ApoEVs were isolated and characterized. In vitro effects of SNTSC-ApoEVs on the proliferation, migration, and tube formation of human umbilical vein endothelial cells (HUVECs) were evaluated by CCK-8, wound healing, transwell, and tube formation assays. The mRNA and protein levels of proangiogenic genes were quantified by qRT-PCR, Western blot, and immunofluorescence analysis. A Matrigel plug model was established in 6-week-old male nu/nu mice for one week, and the in vivo impact of SNTSC-ApoEVs on micro-vessel formation was assessed by histological analysis. Proteomic analysis and RNA sequencing were performed to explore the active ingredients and underlying mechanisms.
RESULTS
SNTSC-ApoEVs enhanced the proliferation, migration, and angiogenesis of HUVECs in vitro. In the Matrigel plug model in vivo, SNTSC-ApoEVs promoted CD31-positive luminal structure formation. Apart from expressing general ApoEV markers, SNTSC-ApoEVs were enriched with multiple proteins related to extracellular matrix-cell interactions. Mechanistically, SNTSC-ApoEVs transferred COL1A1 to HUVECs and promoted endothelial functions by activating the PI3K/Akt/VEGF cascade.
CONCLUSION
SNTSC-ApoEVs can promote angiogenesis by transferring the functional molecule COL1A1 and activating the PI3K/Akt/VEGF pathway, making SNTSC-ApoEVs a promising strategy for the treatment of angiogenesis-related diseases.
目的
血管生成是一个受到严密控制的过程，它启动新血管的形成，其功能障碍可导致危及生命的疾病。凋亡型细胞外囊泡（ApoEVs）作为一种具有高安全性和高分离效率的促血管生成剂而出现，而来源于额外牙源性牙髓干细胞的 ApoEVs（SNTSC-ApoEVs）具有易于获取亲本细胞源和非侵入性细胞采集的独特优势。然而，SNTSC-ApoEVs 的详细特征在很大程度上尚不清楚。本研究旨在探讨 SNTSC-ApoEVs 的促血管生成能力和功能分子。
方法
分离和鉴定 SNTSC-ApoEVs。通过 CCK-8、划痕愈合、Transwell 和管形成试验评估 SNTSC-ApoEVs 对人脐静脉内皮细胞（HUVECs）增殖、迁移和管形成的体外影响。通过 qRT-PCR、Western blot 和免疫荧光分析定量测定促血管生成基因的 mRNA 和蛋白水平。在 6 周龄雄性 nu/nu 小鼠中建立 Matrigel plugs 模型 1 周，通过组织学分析评估 SNTSC-ApoEVs 对微血管形成的体内影响。进行蛋白质组学分析和 RNA 测序以探索活性成分和潜在机制。
结果
SNTSC-ApoEVs 增强了 HUVECs 的体外增殖、迁移和血管生成。在体内 Matrigel plugs 模型中，SNTSC-ApoEVs 促进了 CD31 阳性管腔结构的形成。除了表达一般的 ApoEV 标志物外，SNTSC-ApoEVs 还富含与细胞外基质-细胞相互作用相关的多种蛋白质。从机制上讲，SNTSC-ApoEVs 将 COL1A1 转移到 HUVECs 中，并通过激活 PI3K/Akt/VEGF 级联促进内皮功能。
结论
SNTSC-ApoEVs 通过转移功能分子 COL1A1 并激活 PI3K/Akt/VEGF 途径促进血管生成，使 SNTSC-ApoEVs 成为治疗与血管生成相关疾病的有前途的策略。
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