Horie Shahd, Gaynard Sean, Murphy Mary, Barry Frank, Scully Michael, O'Toole Daniel, Laffey John G
Anaesthesia, School of Medicine, National University of Ireland, Galway, Ireland.
Regenerative Medicine Institute (REMEDI) at CÚRAM Centre for Research in Medical Devices, National University of Ireland Galway, Galway, Ireland.
Intensive Care Med Exp. 2020 Feb 5;8(1):8. doi: 10.1186/s40635-020-0295-5.
Human mesenchymal stem/stromal cells (hMSCs) represent a promising therapeutic strategy for ventilator-induced lung injury (VILI) and acute respiratory distress syndrome. Translational challenges include restoring hMSC efficacy following cryopreservation, developing effective xenogeneic-free (XF) hMSCs and establishing true therapeutic potential at a clinically relevant time point of administration. We wished to determine whether cytokine pre-activation of cryopreserved, bone marrow-derived XF-hMSCs would enhance their capacity to facilitate injury resolution following VILI and elucidate mechanisms of action.
Initially, in vitro studies examined the potential for the secretome from cytokine pre-activated XF-hMSCs to attenuate pulmonary epithelial injury induced by cyclic mechanical stretch. Later, anaesthetised rats underwent VILI and, 6 h following injury, were randomized to receive 1 × 10 XF-hMSC/kg that were (i) naive fresh, (ii) naive cryopreserved, (iii) cytokine pre-activated fresh or (iv) cytokine pre-activated cryopreserved, while control animals received (v) vehicle. The extent of injury resolution was measured at 24 h after injury. Finally, the role of keratinocyte growth factor (KGF) in mediating the effect of pre-activated XF-hMSCs was determined in a pulmonary epithelial wound repair model.
Pre-activation enhanced the capacity of the XF-hMSC secretome to decrease stretch-induced pulmonary epithelial inflammation and injury. Both pre-activated fresh and cryopreserved XF-hMSCs enhanced resolution of injury following VILI, restoring oxygenation, improving lung compliance, reducing lung leak and improving resolution of lung structural injury. Finally, the secretome of pre-activated XF-hMSCs enhanced epithelial wound repair, in part via a KGF-dependent mechanism.
Cytokine pre-activation enhanced the capacity of cryopreserved, XF-hMSCs to promote injury resolution following VILI, potentially via a KGF-dependent mechanism.
人间充质干/基质细胞（hMSCs）是呼吸机诱导性肺损伤（VILI）和急性呼吸窘迫综合征颇具前景的治疗策略。转化过程中的挑战包括冷冻保存后恢复hMSC的疗效、开发有效的无动物源（XF）hMSCs以及在临床相关给药时间点确定其真正的治疗潜力。我们希望确定细胞因子预激活冷冻保存的骨髓来源XF-hMSCs是否会增强其促进VILI后损伤修复的能力，并阐明其作用机制。
首先，体外研究检测细胞因子预激活的XF-hMSCs分泌组减轻周期性机械拉伸诱导的肺上皮损伤的潜力。随后，对麻醉大鼠造成VILI，损伤后6小时将其随机分为接受1×10 XF-hMSC/kg的组，分别为（i）未处理的新鲜细胞、（ii）未处理的冷冻保存细胞、（iii）细胞因子预激活的新鲜细胞或（iv）细胞因子预激活的冷冻保存细胞，而对照动物接受（v）载体。在损伤后24小时测量损伤修复程度。最后，在肺上皮伤口修复模型中确定角质形成细胞生长因子（KGF）在介导预激活的XF-hMSCs作用中的作用。
预激活增强了XF-hMSC分泌组减少拉伸诱导的肺上皮炎症和损伤的能力。预激活的新鲜和冷冻保存的XF-hMSCs均增强了VILI后损伤的修复，恢复了氧合，改善了肺顺应性，减少了肺渗漏并改善了肺结构损伤的修复。最后，预激活的XF-hMSCs分泌组增强了上皮伤口修复，部分通过KGF依赖性机制。
细胞因子预激活增强了冷冻保存的XF-hMSCs促进VILI后损伤修复的能力，可能通过KGF依赖性机制实现。
