Histochemical, immunohistological and scanning electron microscope analysis of tissue retained on spontaneously...

The aim of this study was to analyse the tissue surrounding prematurely extruded ventilation tubes (grommets). Thirty-one ventilation tubes, including 21 Shah and six Shepherd tubes, that had been extruded naturally into the ear canal were examined. After formalin fixation, material adhering to the tubes was retrieved and processed to paraffin wax. Five tubes were processed for scanning electron microscope (SEM) analysis. Sections were stained using histochemical methods for collagen, keratin and keratohyaline. In addition, the presence of collagen (type I and III), keratin, vimentin, fibronectin, tenascin, factor VIII-related antigen, CD31 and CD45 was tested for by immunohistochemistry. Results showed that all specimens consisted of an acellular fibrous material, oriented in concentric rings parallel to the tube surface that was often associated with small collections of CD45+ inflammatory cells. Two specimens contained collagen that was detectable by histochemical and immunohistological methods. Twelve specimens contained identifiable desquamated epithelial cells containing keratin and keratohyaline. Only one specimen stained positively for connective tissue markers (vimentin, fibrous fibronectin, tenascin) or showed the presence of vascular epithelium. SEM revealed adherent clusters or sheets of plate-like structures, consistent with the presence of epithelial squames, on three of the five ventilation tubes examined. It is concluded that tissue retained on extruded grommets consists of orthokeratinitizing epithelium infiltrated by inflammatory cells, a finding that is consistent with the theory that tube extrusion occurs as a result of squamous epithelial proliferation, medial migration, differentiation and desquamation.