From the Department of Surgery (M.B.), Department of Laboratory Medicine (A.T., P.P., B.Y.M., L.R.V., M.K., S.P.), University of California, San Francisco; Vitalant Research Institute (P.P.T., H.T., P.N.), San Francisco, California.
J Trauma Acute Care Surg. 2022 Mar 1;92(3):489-498. doi: 10.1097/TA.0000000000003487.
Hemorrhagic shock and trauma (HS/T)-induced gut injury may play a critical role in the development of multi-organ failure. Novel therapies that target gut injury and vascular permeability early after HS/T could have substantial impacts on trauma patients. In this study, we investigate the therapeutic potential of human mesenchymal stem cells (MSCs) and MSC-derived extracellular vesicles (MSC EVs) in vivo in HS/T in mice and in vitro in Caco-2 human intestinal epithelial cells.
In vivo, using a mouse model of HS/T, vascular permeability to a 10-kDa dextran dye and histopathologic injury in the small intestine and lungs were measured among mice. Groups were (1) sham, (2) HS/T + lactated Ringer's (LR), (3) HS/T + MSCs, and (4) HS/T + MSC EVs. In vitro, Caco-2 cell monolayer integrity was evaluated by an epithelial cell impedance assay. Caco-2 cells were pretreated with control media, MSC conditioned media (CM), or MSC EVs, then challenged with hydrogen peroxide (H2O2).
In vivo, both MSCs and MSC EVs significantly reduced vascular permeability in the small intestine (fluorescence units: sham, 456 ± 88; LR, 1067 ± 295; MSC, 765 ± 258; MSC EV, 715 ± 200) and lung (sham, 297 ± 155; LR, 791 ± 331; MSC, 331 ± 172; MSC EV, 303 ± 88). Histopathologic injury in the small intestine and lung was also attenuated by MSCs and MSC EVs. In vitro, MSC CM but not MSC EVs attenuated the increased permeability among Caco-2 cell monolayers challenged with H2O2.
Mesenchymal stem cell EVs recapitulate the effects of MSCs in reducing vascular permeability and injury in the small intestine and lungs in vivo, suggesting MSC EVs may be a potential cell-free therapy targeting multi-organ dysfunction in HS/T. This is the first study to demonstrate that MSC EVs improve both gut and lung injury in an animal model of HS/T.
出血性休克和创伤(HS/T)引起的肠道损伤可能在多器官衰竭的发展中起关键作用。针对 HS/T 后早期肠道损伤和血管通透性的新型疗法可能对创伤患者产生重大影响。在这项研究中,我们在体内研究了人类间充质干细胞(MSCs)和 MSC 衍生的细胞外囊泡(MSC EVs)在 HS/T 诱导的小鼠中的治疗潜力,并在体外研究了 Caco-2 人肠上皮细胞。
在体内,使用 HS/T 诱导的小鼠模型,测量了血管对 10 kDa 葡聚糖染料的通透性以及小肠和肺的组织病理学损伤。各组为(1)假手术组,(2)HS/T+乳酸林格氏液(LR)组,(3)HS/T+MSC 组和(4)HS/T+MSC EV 组。在体外,通过上皮细胞阻抗测定评估 Caco-2 细胞单层的完整性。用对照培养基、MSC 条件培养基(CM)或 MSC EV 预处理 Caco-2 细胞,然后用过氧化氢(H2O2)处理。
体内,MSC 和 MSC EV 均显著降低了小肠(荧光单位:假手术组 456 ± 88;LR 组 1067 ± 295;MSC 组 765 ± 258;MSC EV 组 715 ± 200)和肺(假手术组 297 ± 155;LR 组 791 ± 331;MSC 组 331 ± 172;MSC EV 组 303 ± 88)的血管通透性。MSC 和 MSC EV 也减轻了小肠和肺的组织病理学损伤。在体外,MSC CM 而非 MSC EV 减轻了 H2O2 处理的 Caco-2 细胞单层通透性的增加。
间充质干细胞 EV 再现了 MSC 降低体内小肠和肺血管通透性和损伤的作用,提示 MSC EV 可能是一种针对 HS/T 中多器官功能障碍的潜在无细胞治疗方法。这是第一项证明 MSC EV 可改善 HS/T 诱导的动物模型中肠道和肺损伤的研究。