Gupta Akash, Burgess Janette K, Slebos Dirk-Jan, Pouwels Simon D
Department of Pulmonology, University Medical Center Groningen, University of Groningen, Groningen, Netherlands.
Department of Pathology and Medical Biology, University Medical Center Groningen, University of Groningen, Groningen, Netherlands.
Front Med (Lausanne). 2023 Jul 26;10:1172622. doi: 10.3389/fmed.2023.1172622. eCollection 2023.
Analysis of respiratory biomarkers or pharmaceutical drug concentrations in bronchial epithelial lining fluid (bELF) using a high-precision sampling method is crucial for effective clinical respiratory diagnostics and research. Here, we utilized a cellulose matrix as an absorptive probe for bELF sampling, subsequently testing the design of a device and sampling technique .
The absorptive matrix [Whatman® qualitative filter paper (Grade -12)] was first tested through tissue-contact experiments on porcine airway tissue. The absorption and elution capacity of the matrix, as well as the laboratory processing and analysis method, was validated with a range of Interleukin-8 (CXCL8) and C-Reactive protein (CRP) stock solutions. Subsequently, the device's design was optimized for universal in-house production and both, safe and efficient sampling. The airway sampling method was then tested in a group of 10 patients with Chronic Obstructive Pulmonary Disease (COPD). For each patient, a bELF sample was obtained using the newly developed bELF probe, as well as a reference 20 mL saline bronchial wash sample. Supernatants were assessed, using an immunoassay, for levels of the pro-inflammatory markers CXCL8, Myeloperoxidase (MPO), and CRP. The bELF samples were compared to bronchial wash.
The Whatman® qualitative filter paper (Grade -12) bELF probes adhered to porcine airway tissue, softening slightly upon wetting. The material maintained architectural integrity following the removal of the probes, leaving no residual fibers on the porcine airway mucosa. The bELF probe design was optimized for bronchoscopic delivery and in-house production. On average, a fully saturated bELF probe carried 32 μL of protein-rich fluid. The mean return of CXCL8 and CRP from samples collected from a serial dilution series (1, 5, 10, 20 ng/mL) was 69% (range 48%-87%). The bELF probe detected, on average, 7 (MPO), 14 (CRP), and 59 (CXCL8) times higher equivalent inflammatory protein concentrations in the collected bELF probe samples compared to the bronchial wash.
The bELF probe is an effective absorptive technology for high-precision bELF sampling without dilution. With a simple in-house production procedure and bronchoscopic sampling technique, this method can be introduced in any bronchoscopic center for a consistent sampling of bELF.
使用高精度采样方法分析支气管上皮衬液（bELF）中的呼吸生物标志物或药物浓度对于有效的临床呼吸诊断和研究至关重要。在此，我们使用纤维素基质作为bELF采样的吸收性探针，随后测试了一种装置的设计和采样技术。
首先通过对猪气道组织进行组织接触实验，对吸收性基质[Whatman®定性滤纸（12级）]进行测试。用一系列白细胞介素-8（CXCL8）和C反应蛋白（CRP）储备溶液验证了该基质的吸收和洗脱能力，以及实验室处理和分析方法。随后，对该装置的设计进行了优化，以便进行通用的内部生产以及安全高效的采样。然后在一组10例慢性阻塞性肺疾病（COPD）患者中测试气道采样方法。对于每位患者，使用新开发的bELF探针以及参考20 mL生理盐水支气管灌洗样本获取bELF样本。使用免疫测定法评估上清液中促炎标志物CXCL8、髓过氧化物酶（MPO）和CRP的水平。将bELF样本与支气管灌洗样本进行比较。
Whatman®定性滤纸（12级）bELF探针粘附在猪气道组织上，湿润后会稍微变软。移除探针后，该材料保持结构完整性，在猪气道黏膜上没有残留纤维。bELF探针设计针对支气管镜递送和内部生产进行了优化。平均而言，一个完全饱和的bELF探针携带32 μL富含蛋白质的液体。从系列稀释系列（1、5、10、20 ng/mL）收集的样本中，CXCL8和CRP的平均回收率为69%（范围48%-87%）。与支气管灌洗相比，bELF探针在收集的bELF探针样本中检测到的等效炎症蛋白浓度平均高出7倍（MPO）、14倍（CRP）和59倍（CXCL8）。
bELF探针是一种有效的吸收技术，可用于高精度的bELF采样且无需稀释。通过简单的内部生产程序和支气管镜采样技术，该方法可在任何支气管镜检查中心引入，以实现bELF的一致采样。
