Gamarra Fernando, Bergner Albrecht, Stauss Elisabeth, Stocker Ingrid, Grundler Stefan, Huber Rudolf Maria
Division of Pneumology, Department of Internal Medicine, University Hospital, Ludwig Maximilian University, Munich, Germany.
Respiration. 2006;73(5):664-72. doi: 10.1159/000092672. Epub 2006 Apr 7.
We evaluated a new in vitro model for mucociliary transport function. Spheroids of human respiratory epithelium show beating cilia at their surface. When cultured in their own mucus, spheroids can rotate along their axis due to coordinated ciliary beating.
To assess whether this setup yields meaningful results we measured rotation frequency (RF) of human bronchial or nasal epithelial spheroids under different temperatures and concentrations of isoproterenol. Isoproterenol was administered either as caged compound releasing active isoproterenol after illumination with UV light, or through a permeable membrane in a two-chamber system.
Under stable conditions, RF remained constant over 200 min. Between 27 and 35 degrees C, there was a temperature-dependent increase: RF(27)( degrees )(C) = 0.27 +/- 0.08 s(-1), and RF(37)( degrees )(C) = 0.43 +/- 0.10 s(-1) (means +/- SEM). Isoproterenol (10(-5), 10(-4) and 10(-3) mmol/l) induced concentration-dependent increases in RF (9, 20 and 25%, respectively; medians) if applied in the two-chamber system. The experiments with caged isoproterenol did not yield conclusive results, probably because the byproducts from photolysis negatively affected ciliary function. The transport velocity at the surface of bronchial and nasal spheroids was estimated to be 2.96 and 3.62 mm/min (medians), respectively, which is in the same range as mucus transport velocity measured in vivo in humans.
This setup can be used to study mucociliary transport function under controlled conditions in vitro, in particular as RF is likely to reflect not only ciliary beat frequency, but also the coordination of ciliary beating and the properties of the mucus.
我们评估了一种用于黏液纤毛运输功能的新型体外模型。人呼吸道上皮球体在其表面显示出跳动的纤毛。当在自身黏液中培养时，球体可因纤毛的协同跳动而沿其轴旋转。
为评估该装置是否能产生有意义的结果，我们测量了在不同温度和异丙肾上腺素浓度下人支气管或鼻上皮球体的旋转频率（RF）。异丙肾上腺素要么作为笼形化合物，在紫外线照射后释放活性异丙肾上腺素，要么通过两室系统中的渗透膜给药。
在稳定条件下，RF在200分钟内保持恒定。在27至35摄氏度之间，存在温度依赖性增加：RF（27）（摄氏度）=0.27±0.08秒-1，RF（37）（摄氏度）=0.43±0.10秒-1（均值±标准误）。如果在两室系统中应用，异丙肾上腺素（10-5、10-4和10-3毫摩尔/升）可诱导RF呈浓度依赖性增加（分别为9%、20%和25%；中位数）。笼形异丙肾上腺素的实验未得出确凿结果，可能是因为光解产生的副产物对纤毛功能产生了负面影响。支气管和鼻球体表面的运输速度估计分别为2.96和3.62毫米/分钟（中位数），这与在人体体内测量的黏液运输速度处于同一范围。
该装置可用于在体外可控条件下研究黏液纤毛运输功能，特别是因为RF可能不仅反映纤毛跳动频率，还反映纤毛跳动的协调性和黏液的特性。
