Centre de Recherche du Centre Hospitalier de l'Université de Montréal (CRCHUM), Montreal, Quebec, Canada; Division of Otorhinolaryngology, Ribeirão Preto Medical School, University of São Paulo (FMRP-USP), Ribeirão Preto, Brazil.
Centre de Recherche du Centre Hospitalier de l'Université de Montréal (CRCHUM), Montreal, Quebec, Canada; Département de Médecine, Université de Montréal, Montreal, Quebec, Canada.
J Allergy Clin Immunol. 2019 Feb;143(2):591-603.e3. doi: 10.1016/j.jaci.2018.05.035. Epub 2018 Jun 20.
The effect of Staphylococcus aureus on nasal epithelial repair has never been assessed in patients with chronic rhinosinusitis with nasal polyps (CRSwNP).
This study aimed to determine whether (1) nasal epithelial cell cultures from patients with CRSwNP and control subjects repair differently; (2) S aureus exoproducts compromise nasal epithelial repair; (3) S aureus alters lamellipodial dynamics; and (4) deleterious effects could be counteracted by the Rho-associated coiled-coil kinase inhibitor Y-27632.
Primary nasal epithelial cells (pNECs) collected during surgeries were cultured and injured under 3 conditions: (1) basal conditions, (2) exposed to S aureus exoproducts, and (3) exposed to S aureus exoproducts and Y-27632. Epithelial repair, lamellipodial dynamics, and cytoskeletal organization were assessed.
Under basal conditions, pNEC cultures from patients with CRSwNP presented significantly lower repair rates and reduced lamellipodial protrusion length and velocity than those from control subjects. S aureus exoproducts significantly decreased repair rates and protrusion dynamics in both control subjects and patients with CRSwNP; however, the effect of S aureus on cell protrusions was more sustained over time in patients with CRSwNP. Under basal conditions, immunofluorescence assays showed significantly reduced percentages of cells with lamellipodia at the wound edge in patients with CRSwNP compared with control subjects. S aureus altered cell polarity and decreased the percentage of cells with lamellipodia in both groups. Finally, Y-27632 prevented the deleterious effects of S aureus exoproducts on CRSwNP repair rates, as well as on lamellipodial dynamics and formation.
S aureus exoproducts significantly alter epithelial repair and lamellipodial dynamics on pNECs, and this impairment was more pronounced in patients with CRSwNP. Importantly, Y-27632 restored epithelial repair and lamellipodial dynamics in the presence of S aureus exoproducts.
金黄色葡萄球菌对慢性鼻-鼻窦炎伴鼻息肉（CRSwNP）患者鼻上皮修复的影响从未被评估过。
本研究旨在确定：（1）CRSwNP 患者和对照鼻上皮细胞培养物的修复是否不同；（2）金黄色葡萄球菌外产物是否会损害鼻上皮修复；（3）金黄色葡萄球菌是否会改变片状伪足动力学；以及（4）Rho 相关卷曲螺旋激酶抑制剂 Y-27632 是否可以对抗有害作用。
在手术期间收集的原代鼻上皮细胞（pNEC）进行培养，并在 3 种条件下进行损伤：（1）基础条件下；（2）暴露于金黄色葡萄球菌外产物下；（3）暴露于金黄色葡萄球菌外产物和 Y-27632 下。评估上皮修复、片状伪足动力学和细胞骨架组织。
在基础条件下，CRSwNP 患者的 pNEC 培养物的修复率明显低于对照组，且片状伪足突起的长度和速度也降低。金黄色葡萄球菌外产物显著降低了对照组和 CRSwNP 患者的修复率和突起动力学；然而，金黄色葡萄球菌对细胞突起的影响在 CRSwNP 患者中随时间推移更为持久。在基础条件下，免疫荧光检测显示，与对照组相比，CRSwNP 患者的伤口边缘具有片状伪足的细胞百分比明显降低。金黄色葡萄球菌改变了细胞极性，并降低了两组细胞的片状伪足百分比。最后，Y-27632 可防止金黄色葡萄球菌外产物对 CRSwNP 修复率、片状伪足动力学和形成的有害影响。
金黄色葡萄球菌外产物显著改变了 pNEC 上的上皮修复和片状伪足动力学，而在 CRSwNP 患者中这种损伤更为明显。重要的是，Y-27632 在存在金黄色葡萄球菌外产物的情况下恢复了上皮修复和片状伪足动力学。
