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Although airway epithelium is known to be modified during chronic respiratory diseases, epithelial cells have rarely been precisely quantified. We therefore intended to evaluate epithelial cell distribution in inflammatory airways, using a cytological approach. Nasal airway cells in 12 patients with nonallergic chronic rhinitis were sampled by brushing, quantified after cytocentrifugation and compared to those from eight controls. Cell populations were quantified after May-Grunwald Giemsa staining and alpha-tubulin immunolabelling to demonstrate ciliary differentiation. When compared to controls, rhinitis patients exhibited lower percentages of ciliated cells (59 +/- 4 versus 32 +/- 2%, respectively), and higher percentages of goblet (24 +/- 3 versus 37 +/- 2%) and basal cells (9 +/- 1 versus 18 +/- 2%). After tubulin immunolabelling, positive staining was specifically detected in cells with cilia (LC+), and in the cytoplasm of some small round cells without obvious cilia (LC-). Fewer immunolabelled cells were detected in rhinitis patients than in controls (with significantly lower percentages of LC+ and higher percentages of LC-). Nasal brushing is an effective technique for quantification of airway epithelial cells. Tubulin immunolabelling is useful to detect ciliated cells and distinguishes another cell population, possibly preciliated cells. These cytological findings suggest the presence of modifications of epithelial differentiation and proliferation, possibly related to local chronic inflammation.

The modifications of epithelial differentiation and proliferation observed in nasal polyps (NP) could be related to local secretion of growth factors, among which platelet-derived growth factor (PDGF) could play a key role. We therefore prospectively studied, by immunohistochemistry, proliferating cell nuclear antigen (PCNA, an S-phase cell marker), PDGF, and CD-68 (activated macrophages marker) expression in NP and inferior turbinate mucosa (NM) in 11 patients. Our data show that PCNA and PDGF expression are increased in NP epithelium, while CD-68 expression is increased in NP epithelium and lamina propria when compared to NM. Increased local PDGF secretion by numerous activated macrophages could therefore be involved in epithelial cell proliferation up-regulation in NP. PDGF could also be involved in the pathogenesis of NP via its connective tissue remodeling actions.

Objective: To detect, quantify, and compare respiratory epithelial cell proliferation in nasal mucosa and polyps from patients with nasal polyposis. Design: Cohort study. Setting: Patients and samples were selected at the Hopital Intercommunal de Creteil (France). Flow cytofluorometry and immunohistochemistry were performed at Hopitaux Tenon and Mondor (Universite Paris [France] VI et XII). Patients: Twenty-one patients undergoing endoscopic ethmoidectomy for treatment of nasal polyposis. Methods: In 10 cases, epithelial cells were removed from frozen inferior turbinate mucosa and polyps by mechanical disaggregation and were then analyzed by flow cytofluorometry, providing the cell DNA content (propidium iodide labeling) and the percentage of S-phase cells. In 11 cases, inferior turbinate mucosa and polyps were fixed in formaldehyde and embedded in paraffin. Proliferating cell nuclear antigen expression in the epithelium was quantified by immunohistochemistry: a proliferating cell nuclear antigen index was calculated for each sample in the basal area, suprabasal area, and full height of the epithelium. Results: All cell populations studied were diploid, and percentages of S-phase cells were significantly higher in nasal polyps than in mucosa. Proliferating cell nuclear antigen indexes were significantly higher in nasal polyps than in the suprabasal area and full height of the mucosal epithelium. Conclusion: Cell proliferation is increased in epithelium from nasal polyps. Epithelial damage caused by inflammatory mediators could induce this increased cell proliferation via epithelial repair processes. Inflammatory cells could up-regulate epithelial cell proliferation by secreting growth factors. (Arch Otolaryngol Head Neck Surg. 1996;122:432-436)

Analysis of nasal cytology provides useful information for the diagnosis of rhinitis. This analysis represents an interesting tool for objective assessment of nasal pathology. The purpose of this study was to describe the brushing method for obtaining nasal cytologic specimen. Samples were carried out from nine adult patients with chronic nasal obstruction. In all cases, the cytologic specimen had a good amount of well-preserved cells, easy to identify on morphological criteria. A differential cell count was performed on the different epithelial cell types present in the specimen and the results are reported in the present study. In conclusion, brushing method is non invasive and allows to precisely evaluate both epithelial and inflammatory cells present in the nasal mucosa.

The cytological study of the nasal mucosa is one the essential steps toward a better understanding of the physicopathological mechanisms involved in chronic affections of the nose. The development of a reliable, reproducible noninvasive technique to obtain cells in the prerequisite for this analysis. The brush technique is based on the use of a small cylindric nylon brush which is rotated as it is moved back and forth across the nasal mucosa to harvest the various cell types present on the surface of the mucosa. The specimen obtained is centrifuged and examined under an optical microscope. A semi-quantitative analysis is carried out to determine the relative richness of the specimen in epithelial cells (ciliated, qoblet and basal cells) and in inflammatory cells (neutrophils, basophils, eosinophils, lymphocytes, and monocytes). A precise differential cell count is then performed. According to the results obtained, an immunologic marker study of lymphocyte subsets may also be done using the same specimen. This technique may be used in both children and adults and permits cell harvest from various sites within the nasal fossa. Already tested in the study of the activity of cilia, this method is promising for the cytological study of the nasal mucosa. The brush method and other techniques of cell harvest are described and compared.