Controlled Photo-Release of Nitric Oxide for Antimicrobial Inhalation Therapy

一氧化氮(NO)是一种强有力的内源性抗菌/抗病毒药物，通常存在于中等水平 (200-1000 ppbv)在健康患者的上呼吸道/鼻窦内，有助于预防慢性上呼吸道 感染。患有慢性鼻窦炎(CRS)等难治性疾病的患者较低 呼吸道感染--慢性阻塞性肺疾病(COPD)、囊性纤维化(CF)和其他--可能 可能受益于家庭吸入性NO疗法(INO)。而iNO的水平要高得多(10- 50ppmv)是医院治疗新生儿肺动脉高压和成人急性呼吸道疾病的常规药物。 窘迫综合症，目前iNO系统的极高成本(每天3,000美元)使其无法用于 慢性阻塞性肺疾病、慢性阻塞性肺疾病和慢性阻塞性肺疾病相关呼吸道感染的家庭治疗对于这样的障碍， 给药剂量模拟健康人上呼吸道正常水平的NO 可能是一种安全有效的防治上下呼吸道感染的方法。在这里， 我们建议研究一种新的低成本递送策略来产生用于iNO治疗的纯一氧化氮(NO)，该策略 可在医院或家中用于某些没有标准慢性治疗的临床情况 当前存在(例如，慢性阻塞性肺病、慢性阻塞性肺病和慢性阻塞性肺病)。我们假设通过封装稳定的非捐赠者S- 亚硝基-N-乙酰青霉胺(SNAP)，进入普通聚合物管中进行iNO治疗，光激活释放 可以实现在载体空气中产生受控的治疗性NO水平。为了检验这一假设，在 目的1我们将表征从装有SNAP的管路向气流中释放光活性NO的变量。 我们将SNAP装入聚合管内，启动时环境空气将通过管路输送 用泛光灯进行光解NO释放。将对NO进行监测，以评估配方变量在 卡扣式油管会影响光解NO释放的水平和寿命。在目标2中，我们将结合LED 光和新的NO传感器，带有来自Aim 1的最佳SNAP加载管材配方，以监测和 达到治疗性一氧化氮水平。我们将测试不同的加湿空气流量与最佳管型、 几何形状和SNAP负荷可以在200-2000的目标范围内提供持续、稳定的NO水平 在气流中保持至少10小时的ppbv，使用传感器信号控制LED强度。在《目标3》中，我们将 使用目标2中设计的系统检查iNO受控传递的体外杀菌能力。我们会 培养金黄色葡萄球菌和金黄色葡萄球菌的生物膜，并将它们暴露于不同时间的特定iNO水平 带湿空气的200-2000 ppbv范围。生物膜生物量和活细菌将被确定为评估 SNAP加载油管产生的INO效应。人气管上皮细胞生长在半透膜上 气/液界面的膜也将用iNO方法处理，以证明产生的NO 水平对这些细胞没有不良影响。SNAP负荷对不同NO水平的影响 还将检查生长在人类上皮细胞表面的生物膜上的管状结构。

Nitric oxide (NO) is a potent and endogenous antimicrobial/antiviral agent normally present at moderate levels (200-1000 ppbv) within the upper airways/sinuses of healthy patients, helping to prevent chronic upper airway infections. Patients suffering from chronic rhinosinusitis (CRS) and other conditions with difficult-to-treat lower respiratory infections—chronic obstructive pulmonary disease (COPD), cystic fibrosis (CF) and others—could potentially benefit from home treatment with inhaled NO therapy (iNO). While iNO at much higher levels (10- 50 ppmv) is used routinely in hospitals for neonatal pulmonary hypertension and adult acute respiratory distress syndrome, the extremely high cost of current iNO systems ($3,000 per day) precludes their use for home treatment of respiratory tract infections associated with CRS, COPD and CF. For such disorders, administration of doses of NO that mimic levels normally found in the upper airways of healthy individuals could be a safe and effective method of preventing or treating upper and lower respiratory infection. Herein, we propose to study a new low-cost delivery strategy to generate pure nitric oxide (NO) for iNO therapy, which could be used in the hospital or at home for certain clinical situations where no standard chronic treatment currently exists (e.g., CF, COPD, and CRS). We hypothesize that by encapsulating the stable NO-donor, S- nitroso-N-acetylpenicillamine (SNAP), into common polymeric tubing for iNO therapy, light-activated release of NO can be achieved to yield controlled therapeutic NO levels in the carrier air gas. To test this hypothesis, in Aim 1 we will characterize the variables of light active NO release from SNAP-loaded tubing into an airstream. We will load SNAP into polymeric tubing and ambient air will be delivered through the tubing while initiating photolytic NO release with a flood lamp. NO will be monitored to assess how formulation variables in the SNAP-loaded tubing influence the levels and lifetime of photolytic NO release. In Aim 2, we will combine LED light and a new NO sensor with the optimal SNAP-loaded tubing formulations from Aim 1 to monitor and achieve therapeutic NO levels. We will test how varying humidified air flow rates with optimal tubing type, geometry and SNAP loading can provide continuous, stable levels of NO within the target range of 200-2000 ppbv within an air stream for at least 10 h, using sensor signals to control LED intensity. In Aim 3, we will examine controlled iNO delivery using the system devised in Aim 2 for its ability to kill bacteria in vitro. We will grow biofilms of P. aureginosa and S. aureus and expose them for various times to specific iNO levels in a range of 200-2000 ppbv with humidified air. Biofilm biomass and viable bacteria will be determined to assess iNO effect derived from the SNAP-loaded tubing. Human tracheal epithelial cells grown on a semipermeable membrane at the air/liquid interface will also be treated with the iNO approach to prove that the produced NO levels have no adverse effects on these cells. The effect of various NO levels produced from the SNAP-loaded tubing on biofilms grown on the surface of the human epithelial cells will also be examined.