1 Faculty of Pharmacy, The University of Sydney , Sydney, Australia .
2 Department Pathology and Medical Biology, University Medical Center Groningen, University of Groningen , Groningen, The Netherlands .
J Aerosol Med Pulm Drug Deliv. 2016 Dec;29(6):514-524. doi: 10.1089/jamp.2016.1289. Epub 2016 May 17.
Current treatment regimens for inhalation injury are mainly supportive and rely on self-regeneration processes for recovery. Cell therapy with mesenchymal stromal cells (MSCs) is increasingly being investigated for the treatment of inhalation injury. Human amniotic MSCs (hAMSCs) were used in this study due to their potential use in inflammatory and fibrotic conditions of the lung. This study aimed at demonstrating that hAMSCs can be atomized with high viability, for the purpose of achieving a more uniform distribution of cells throughout the lung. Another aim of this study was to set ground for future application to healthy and diseased lungs by demonstrating that hAMSCs were able to survive after being sprayed onto substrates with different stiffness.
Two methods of atomization were evaluated, and the LMA MAD780 device was selected for atomizing hAMSCs for optimized delivery. To mimic the stiffness of healthy and diseased lungs, gelatin gel (10% w/v) and tissue culture plastic were used as preliminary models. Poly-l-lysine (PLL) and collagen I coatings were used as substrates on which the hAMSCs were cultured after being sprayed.
The feasibility of atomizing hAMSCs was demonstrated with high cell viability (81 ± 3.1% and 79 ± 11.6% for cells sprayed onto plastic and gelatin, respectively, compared with 85 ± 4.8% for control/nonsprayed cells) that was unaffected by the different stiffness of substrates. The presence of the collagen I coating on which the sprayed cells were cultured yielded higher cell proliferation compared with both PLL and no coating. The morphology of sprayed cells was minimally compromised in the presence of the collagen I coating.
This study demonstrated that hAMSCs are able to survive after being sprayed onto substrates with different stiffness, especially in the presence of collagen I. Further studies may advance the effectiveness of cell therapy for lung regeneration.
目前吸入性损伤的治疗方案主要是支持性的，依赖于自身的再生过程来恢复。间充质基质细胞（MSCs）的细胞治疗越来越多地被用于吸入性损伤的治疗。本研究使用人羊膜间充质基质细胞（hAMSCs），因为它们在肺部炎症和纤维化条件下具有潜在的应用价值。本研究旨在证明 hAMSCs 可以以高活力进行雾化，以便更均匀地分布细胞在整个肺部。本研究的另一个目的是通过证明 hAMSCs 在喷洒到具有不同硬度的基质上后能够存活，为未来在健康和患病肺部的应用奠定基础。
评估了两种雾化方法，并选择 LMA MAD780 设备对 hAMSCs 进行雾化，以实现最佳的输送效果。为了模拟健康和患病肺部的硬度，使用明胶凝胶（10%w/v）和组织培养塑料作为初步模型。多聚赖氨酸（PLL）和胶原蛋白 I 涂层被用作喷洒后的 hAMSCs 培养的基质。
高细胞活力证明了雾化 hAMSCs 的可行性（分别喷洒到塑料和明胶上的细胞活力为 81±3.1%和 79±11.6%，而对照组/未喷洒的细胞活力为 85±4.8%），这不受基质硬度不同的影响。喷洒细胞在胶原蛋白 I 涂层上培养时，细胞增殖率高于 PLL 和无涂层。在胶原蛋白 I 涂层存在的情况下，喷洒细胞的形态受到的影响最小。
本研究表明，hAMSCs 能够在喷洒到具有不同硬度的基质上后存活，尤其是在存在胶原蛋白 I 的情况下。进一步的研究可能会提高细胞治疗肺部再生的效果。
