Department of Surgery, University of California, San Francisco, 513 Parnassus Ave, San Francisco, CA, 94143, USA.
Department of Laboratory Medicine, University of California, San Francisco, 513 Parnassus Ave , San Francisco, CA, 94143, USA.
J Transl Med. 2024 Jun 21;22(1):588. doi: 10.1186/s12967-024-05406-1.
Patients with hemorrhagic shock and trauma (HS/T) are vulnerable to the endotheliopathy of trauma (EOT), characterized by vascular barrier dysfunction, inflammation, and coagulopathy. Cellular therapies such as mesenchymal stem cells (MSCs) and MSC extracellular vesicles (EVs) have been proposed as potential therapies targeting the EOT. In this study we investigated the effects of MSCs and MSC EVs on endothelial and epithelial barrier integrity in vitro and in vivo in a mouse model of HS/T. This study addresses the systemic effects of HS/T on multiorgan EOT.
In vitro, pulmonary endothelial cell (PEC) and Caco-2 intestinal epithelial cell monolayers were treated with control media, MSC conditioned media (CM), or MSC EVs in varying doses and subjected to a thrombin or hydrogen peroxide (HO) challenge, respectively. Monolayer permeability was evaluated with a cell impedance assay, and intercellular junction integrity was evaluated with immunofluorescent staining. In vivo, a mouse model of HS/T was used to evaluate the effects of lactated Ringer's (LR), MSCs, and MSC EVs on endothelial and epithelial intercellular junctions in the lung and small intestine as well as on plasma inflammatory biomarkers.
MSC EVs and MSC CM attenuated permeability and preserved intercellular junctions of the PEC monolayer in vitro, whereas only MSC CM was protective of the Caco-2 epithelial monolayer. In vivo, both MSC EVs and MSCs mitigated the loss of endothelial adherens junctions in the lung and small intestine, though only MSCs had a protective effect on epithelial tight junctions in the lung. Several plasma biomarkers including MMP8 and VEGF were elevated in LR- and EV-treated but not MSC-treated mice.
In conclusion, MSC EVs could be a potential cell-free therapy targeting endotheliopathy after HS/T via preservation of the vascular endothelial barrier in multiple organs early after injury. Further research is needed to better understand the immunomodulatory effects of these products following HS/T and to move toward translating these therapies into clinical studies.
患有出血性休克和创伤（HS/T）的患者易患创伤性内皮病（EOT），其特征为血管屏障功能障碍、炎症和凝血病。间充质干细胞（MSCs）和 MSC 细胞外囊泡（EVs）等细胞疗法已被提议作为针对 EOT 的潜在治疗方法。在这项研究中，我们在 HS/T 小鼠模型中研究了 MSC 和 MSC EV 对体外和体内内皮和上皮屏障完整性的影响。这项研究解决了 HS/T 对多器官 EOT 的全身影响。
在体外，用对照培养基、MSC 条件培养基（CM）或 MSC EV 以不同剂量处理肺内皮细胞（PEC）和 Caco-2 肠上皮细胞单层，并分别用凝血酶或过氧化氢（HO）进行处理。用细胞阻抗测定法评估单层通透性，并用免疫荧光染色评估细胞间连接的完整性。在体内，使用 HS/T 小鼠模型评估乳酸林格氏液（LR）、MSCs 和 MSC EV 对肺和小肠内皮和上皮细胞间连接以及血浆炎症生物标志物的影响。
MSC EVs 和 MSC CM 可减轻 PEC 单层的通透性并维持其细胞间连接，而仅 MSC CM 对 Caco-2 上皮单层具有保护作用。在体内，MSC EVs 和 MSCs 均减轻了肺和小肠内皮细胞黏附连接的丢失，但只有 MSCs 对肺上皮紧密连接具有保护作用。几种血浆生物标志物，包括 MMP8 和 VEGF，在 LR 和 EV 处理的小鼠中升高，但在 MSC 处理的小鼠中没有升高。
总之，MSC EVs 可作为一种潜在的无细胞疗法，通过在损伤后早期维持多个器官的血管内皮屏障，针对 HS/T 后的内皮病。需要进一步研究以更好地了解这些产品在 HS/T 后的免疫调节作用，并将这些疗法推向临床研究。
