The development of nasal polyp disease involves early nasal mucosal inflammation and remodelling

Juan Meng, Peng Zhou, Yafeng Liu, Feng Liu, Xuelian Yi, Shixi Liu, Gabriele Holtappels, Claus Bachert, Nan Zhang, Juan Meng, Peng Zhou, Yafeng Liu, Feng Liu, Xuelian Yi, Shixi Liu, Gabriele Holtappels, Claus Bachert, Nan Zhang

Abstract

Chronic rhinosinusitis with nasal polyps (CRSwNP) is characterized by both a chronic inflammation and tissue remodelling; as indicated by extracellular matrix protein deposition, basement membrane thickening, goblet cell hyperplasia and subepithelial edema, with reduced vessels and glands. Although remodelling is generally considered to be consequence of persistent inflammation, the chronological order and relationship between inflammation and remodelling in polyp development is still not clear. The aim of our study was therefore to investigate the pathological features prevalent in the development of nasal polyps and to elucidate the chronological order and relationship between inflammation and remodelling, by comparing specific markers of inflammation and remodelling in early stage nasal polyps confined to the middle turbinate (refer to as middle turbinate CRSwNP) obtained from 5 CRSwNP patients with bilateral polyposis, mature ethmoidal polyps from 6 CRSwNP patients, and normal nasal mucosal tissue from 6 control subjects. Middle turbinate CRSwNP demonstrated significantly more severe epithelial loss compared to mature ethmoidal polyps and normal nasal mucosa. The epithelial cell junction molecules E-cadherin, ZO-1 and occludin were also expressed in significantly lower amounts in mature ethmoidal polyps compared to healthy mucosa. Middle turbinate CRSwNP were further characterized by significantly increased numbers of subepithelial eosinophils and M2 type macrophages, with a distinct lack of collagen and deposition of fibronectin in polyp part. In contrast, the turbinate area of the middle turbinate CRSwNP was characterized by an increase in TGF-β activated myofibroblasts expressing α-SMA and vimentin, an increase in the number of pSmad2 positive cells, as well as increased deposition of collagen. These findings suggest a complex network of processes in the formation of CRSwNP; including gross epithelial damage and repair reactions, eosinophil and macrophage cell infiltration, and tissue remodelling. Furthermore, remodelling appears to occur in parallel, rather than subsequent to inflammation.

Conflict of interest statement

Competing Interests: The authors have declared that no competing interests exist.

Figures

Figure 1. Diagram of middle turbinate polyps…
Figure 1. Diagram of middle turbinate polyps in patients with CRSwNP.
As the diagram shows, for the samples of middle turbinate CRSwNP, special attention was paid to the collection of the whole polyp with the stalk and parts of the surrounding mucosa. This allowed us to discriminate into polyp area and turbinate area for analysis.
Figure 2. Epithelial loss (%) in control,…
Figure 2. Epithelial loss (%) in control, mature CRSwNP and middle turbinate CRSwNP.
MT, middle turbinate.
Figure 3. Immunostaining of E-cadherin, ZO-1 and…
Figure 3. Immunostaining of E-cadherin, ZO-1 and occludin in control and mature CRSwNP.
E-cadherin (A, B), ZO-1(C, D), occludin (E, F) for control (A, C, E) and mature CRSwNP (B, D, F).Original magnification 400x.
Figure 4. Quantification of the staining intensity…
Figure 4. Quantification of the staining intensity for E-cadherin, ZO-1 and occludin in control and mature CRSwNP.
Figure 5. Quantification of celluar and extracelluar…
Figure 5. Quantification of celluar and extracelluar components.
Eosinophils(A), CD68-(B), MMR-(C), α-SMA-(D), vimentin-(E), and pSmad2(F) positive cells, Fn (G) and collagen(H) in control interior turbinates (control IT), mature CRSwNP and middle turbinate CRSwNP, separated into turbinate and polyp areas.
Figure 6. Immunostaining of celluar and extracelluar…
Figure 6. Immunostaining of celluar and extracelluar components.
HE (A1-A5), CD68 (B1-B5),MMR (C1-C5), α-SMA (D1-D5), vimentin (E1-E5), pSmad2 (F1-F5), Fn (G1-G5) picrosirius red staining viewed in bright-field (H1-H5) and viewed under polarized light (I1-I5) in mature CRSwNP (A1-G1), polyp area of middle turbinate CRSwNP (A2-G2),turbinate area of middle turbinate CRSwNP (A4-G4) and control mucosa (A5-G5)(final magnification 400×). The overview of middle turbinate CRSwNP was shown in A3-G3 (composed by Photoshop software with pictures in 40×magnification).
Figure 7. α-SMA expression in CRSwNP fibroblasts…
Figure 7. α-SMA expression in CRSwNP fibroblasts as measured by flow cytometry.
Without TGF-β1 stimulation (A), with TGF-β1 stimulation (B), green marks α-SMA positive cells and red marks α-SMA negative cells.
Figure 8. Fn mRNA expression and Fn…
Figure 8. Fn mRNA expression and Fn protein concentration.

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