Nitric oxide diffusion through cystic fibrosis-relevant media and lung tissue
Nitric oxide diffusion through cystic fibrosis-relevant media and lung tissue
复制标题
DOI：
10.1039/c9ra07367a
复制
发表时间：
2019-12-02
期刊：
影响因子：
3.9
通讯作者：
Schoenfisch, Mark H.
中科院分区：
文献类型：
--
作者：
Hall, Jackson R.;Maloney, Sara E.;Schoenfisch, Mark H.
采用简化的扩散池方法来测量一氧化氮 (NO) 通过磷酸盐缓冲盐水 (PBS) 和人工痰液介质 (ASM)（气道粘液的体外类似物）的扩散系数。与 PBS 相比，通过蛋白质 ASM 的扩散产生了显着较低的扩散系数，这归因于粘蛋白网的物理阻碍和 NO 自由基对 ASM 中生物化合物的反应性质。为了进一步证实 ASM 限制 NO 自由扩散，采用大分子丙胺修饰的环糊精供体 (CD-PA) 来更缓慢地释放 NO。相对于 PBS，ASM 中通过 NO 供体的 NO 扩散特性也较慢。由于NO在穿过粘液屏障后可能与肺细胞相互作用，因此在有和没有ASM层的情况下研究了NO和CD-PA大分子NO供体通过分化肺组织的扩散。通过三个扩散屏障的 NO 扩散比较表明，与 PBS 和 ASM 相比，肺组织在实验过程中显着阻碍了 NO 渗透。事实上，CD-PA 通过肺组织的扩散受到阻碍，直到释放其 NO 有效负载后，这可能是由于 NO 供体结构的净电荷增加所致。重要的是，在杀菌浓度下释放 NO 的 CD-PA 不会影响组织的活力。
A simplified diffusion cell methodology was employed to measure the diffusion coefficient of nitric oxide (NO) through phosphate buffered saline (PBS) and artificial sputum medium (ASM)-an in vitro analog for airway mucus. Diffusion through the proteinaceous ASM yielded a significantly lower diffusion coefficient compared to PBS, which is attributed to both the physical obstruction by the mucin mesh and reactive nature of NO radicals towards the biological compounds in ASM. To further confirm that ASM was restricting NO from diffusing freely, a macromolecular propylamine-modified cyclodextrin donor (CD-PA) was employed to release the NO more slowly. The NO diffusion characteristics in ASM via the NO donor were also slower relative to PBS. As NO is likely to interact with lung cells after passing through the mucus barrier, the diffusion of both NO and the CD-PA macromolecular NO donor through differentiated lung tissue was investigated with and without an ASM layer. Comparison of NO diffusion through the three diffusion barriers indicated that the lung tissue significantly impeded NO penetration over the course of the experiment compared to PBS and ASM. In fact, the diffusion of CD-PA through the lung tissue was hindered until after the release of its NO payload, potentially due to the increased net charge of the NO donor structure. Of importance, the viability of the tissue was not influenced by the NO-releasing CD-PA at bactericidal concentrations.