bromochloroacetic-acid and Otitis-Media

Canine aural cholesteatoma is an epidermoid cyst that forms in the middle ear cavity as a rare complication of otitis media but the aetiopathogenesis remains controversial. In the present study, 13 cases of canine aural cholesteatoma were investigated histologically and immunohistochemically and compared with cases of chronic otitis. The immunohistochemical investigation was performed using the following monoclonal antibodies: anti-cytokeratins (CK) 14, 16, 8/18, and 19, and anti-Ki67. The proliferative indexes (PIs) of cholesteatomata and otitis epithelium were calculated as the percentage of Ki67 positive nuclei/total nuclei. Histologically, the cholesteatomata were composed of a hyperplastic, hyperkeratotic epithelium (matrix) resting on a fibrous perimatrix, infiltrated by inflammatory cells and devoid of cutaneous adnexa. Immunohistochemically, the cholesteatoma epithelium was CK14- and CK16-positive, and CK8/18- and CK19-negative. A similar pattern of CK expression was found in otitis externa. In otitis media, ciliated epithelium stained CK8/18- and CK19-positive in all layers, CK14-positive in the basal layers, and CK16-negative. The mean PIs in cholesteatomata and otitides were 18.8 and 17.8, respectively. The immunohistochemical pattern of CK expression in cholesteatomata, when compared with chronic otitis, was suggestive of hyperproliferative epithelium, but its origin could not be demonstrated. Comparable PI values were obtained in cholesteatoma and in chronic otitis, which confirmed that Ki67 is a valuable indicator of a hyperproliferative state, but not a predictor of aggressiveness.

Topics: Animals; Cholesteatoma, Middle Ear; Dog Diseases; Dogs; Ear Canal; Female; Immunohistochemistry; Keratins; Ki-67 Antigen; Male; Otitis Externa; Otitis Media

Established middle-ear cleft cholesteatoma is associated with keratinous debris, which is likely to be an ideal medium for saprophytic fungal colonisation. This prospective case study aimed to explore the incidence and nature of fungal elements in cholesteatoma keratin samples obtained during primary mastoid surgery.. All cases of middle-ear cleft cholesteatoma treated with primary mastoid surgery at the El-Sahel Teaching Hospital over a seven-month period were included. Keratinous debris obtained from the mastoid antrum was subjected to mycological analysis at the Department of Medical Microbiology and Immunology, Faculty of Medicine, Cairo University. A literature search was performed to determine the clinical and pathological relevance of fungal colonisation in cholesteatoma.. Eighteen patients underwent primary mastoid surgery for cholesteatoma (nineteen ears in total) in a seven-month period starting 30 March 2013. Patients included 13 males and 5 females, with an age range of 9 to 45 years (mean 23 years). Fungal cultures were obtained from 17 keratin samples (89 per cent). Of these, five fungal isolates belonged to the dermatophyte group (21 per cent).. Fungal colonisation in middle-ear cleft cholesteatoma probably plays a significant role in disease progression. Moreover, saprophytic fungal colonisation in cholesteatoma keratin may be responsible for the fetor commonly associated with the ear discharge.

Topics: Adolescent; Adult; Child; Cholesteatoma, Middle Ear; Chronic Disease; Disease Progression; Female; Fungi; Granulation Tissue; Humans; Keratins; Male; Mastoid; Middle Aged; Otitis Media; Prospective Studies; Young Adult

Inflammatory conditions of the ear, otitis media, are one of the most common disease entities in children. In this study, the role of the plasminogen (plg)/plasmin system for the spontaneous development of chronic otitis media was investigated by the analysis of plg-deficient mice. Whereas essentially all of the wild-type control mice kept a healthy status of the middle ear, all the plg-deficient mice gradually developed chronic otitis media with various degrees of inflammatory changes during an 18-week observation period. Five bacterial strains were identified in materials obtained from the middle ear cavities of six plg-deficient mice. Morphological studies revealed the formation of an amorphous mass tissue and inflammatory changes in the middle ears of plg-deficient mice. Immunohistochemical studies further indicate a mass infiltration of neutrophils and macrophages as well as the presence of T and B cells in the middle ear mucosa of these mice. Extensive fibrin deposition and an abnormal keratin formation were also observed in the tympanic membrane, the middle ear cavity and external ear canal in these mice. These results suggest that plg plays an essential role in protecting against the spontaneous development of chronic otitis media. Our findings also suggest the possibility of using plg for clinical therapy of certain types of otitis media.

Topics: Animals; B-Lymphocytes; Bacteria; Disease Models, Animal; Ear, External; Ear, Middle; Fibrin; Hematologic Diseases; Immunohistochemistry; Keratins; Macrophages; Mice; Mice, Inbred C57BL; Mice, Inbred DBA; Mucous Membrane; Neutrophil Infiltration; Otitis Media; Plasminogen; T-Lymphocytes; Tympanic Membrane

Rat middle ear epithelial cells were infected with the adeno 12-SV40 hybrid virus. The cell line thus obtained displays features of primary cultured epithelial cells in both light microscopic and ultrastructural examinations. The immortalized cells have been in continuous proliferation for 40 passages and more than 17 months. Immunohistochemical analysis of the immortalized cells was positive for the SV40 T antigen and the tumor suppressor protein p53. The cells also stained positive for cytokeratin, an epithelial cell marker, and negative for vimentin, a fibroblast marker. These results, together with karyotype analysis, indicate that this cell line originated from rat middle ear epithelial cells and retains the characteristics of epithelial cells. This cell line will be useful for studying the normal cellular biology of middle ear epithelial cells, as well as the cellular and molecular mechanisms involved in the bacteria-middle ear epithelial cell interaction.

Topics: Animals; Antigens, Polyomavirus Transforming; Blotting, Western; Cell Culture Techniques; Cell Line; Disease Models, Animal; Ear, Middle; Epithelium; Immunohistochemistry; Karyotyping; Keratins; Male; Microscopy, Electron, Scanning; Otitis Media; Rats; Rats, Wistar; Simian virus 40; Tumor Suppressor Protein p53

The distribution of collagen types I, III, and IV and of laminin, fibronectin, and keratin was studied in otitis media experimentally induced by Streptococcus pneumoniae in the chinchilla. The expression of interstitial collagen types I and III and of fibronectin was increased in the subepithelial space that was thickened by inflammation in the acute period of infection. The expression of collagen type IV in the subepithelial space could be seen in the early period. The epithelial cells in the middle ear changed from flat cuboidal to pseudostratified columnar in pneumococcus-inoculated ears, and the number of keratin-positive epithelial cells in the middle ear increased remarkably after infection. These results indicate that changes in epithelial cell differentiation effected by the extracellular matrix correlate with changes in expression of keratin. It is proposed that the extracellular matrix may contribute to tissue repair in the middle ear after bacterial infection by interfering with cell proliferation of epithelial cells and fibroblasts.

Topics: Animals; Cell Differentiation; Chinchilla; Collagen; Ear, Middle; Epithelial Cells; Extracellular Matrix; Extracellular Matrix Proteins; Fibronectins; Keratins; Mucous Membrane; Otitis Media; Streptococcal Infections; Streptococcus pneumoniae

By introducing certain irritants into the middle ear it is possible to produce cholesteatoma. Propylene glycol, the main agent used for this purpose, produces a long-standing inflammation that causes hyperplasia and migration of the epithelium through an intact tympanic membrane. In the present investigation topical prednisolone was used in order to inhibit the production of cholesteatoma. The results indicate that there is a marked decrease in inflammation and hence experimental cholesteatoma production when prednisolone is administered into the middle ear.

Topics: Animals; Anti-Inflammatory Agents; Cell Movement; Cholesteatoma, Middle Ear; Ear, Middle; Epithelium; Fibrosis; Glucocorticoids; Granulation Tissue; Hyperplasia; Inflammation; Injections; Irritants; Keratins; Otitis Media; Prednisolone; Propylene Glycol; Rats; Rats, Wistar; Temporal Bone; Tympanic Membrane

The presence of keratin in the middle ear cavity is usually associated with the diagnosis of cholesteatoma or epidermoid metaplasia. Analysis of a series of 18 cases suggests that it may correspond to a specific entity developing in the course of severe or long-lasting opened chronic otitis. This condition, we called mallear epidermosis, is characterized by: i) a perforation of the tympanic membrane lining the handle of the malleus and the umbo; ii) a proliferation of keratin surrounding the handle of the malleus and diffusing into the mesotympanum on the internal side of the tympanic membrane, without matrix; and iii) a mild inflammation of the middle ear epithelium. The process is usually limited to the mesotympanum and does not extend towards the attic and the posterior cavities. Epidemiological, clinical, pathophysiological, and histological features allow this entity to be distinguished from cholesteatoma and epidermoid metaplasia. Management is either medical consisting of local treatment and microaspiration, or surgical including resection of the umbo, removal of the tympanic membrane invaded by the adherent hyperkeratotic layers and repair by conventional underlay myringoplasty. This report emphasizes the need for a clear identification of the various types of chronic otitis media presenting with keratin in the middle ear, as they do not share the same course and do not require the same therapeutic management.

Topics: Adolescent; Adult; Aged; Cholesteatoma, Middle Ear; Chronic Disease; Diagnosis, Differential; Ear, Middle; Epithelium; Female; Humans; Keratins; Male; Malleus; Metaplasia; Middle Aged; Myringoplasty; Otitis Media; Suction; Tympanic Membrane; Tympanic Membrane Perforation

The nature and origin of the epithelial layers in acquired cholesteatoma is still unclear. Although previous morphological studies comparing external meatal and cholesteatoma epithelium have shown no significant difference, bone resorption is generally much more severe with cholesteatoma than with chronic otitis media without cholesteatoma. It is possible that cholesteatoma epithelium has undergone transformation leading to its enhanced bone destroying role. In this study the cytokeratin patterns of aural and cholesteatoma epithelia grown in cell culture were compared using monoclonal antibodies. No significant difference in staining patterns were found suggesting that there has been no change in cell phenotype which maintains that of external auditory meatus epithelium. This study therefore supports the immigration theory of cholesteatoma genesis.

Topics: Antibodies, Monoclonal; Bone Resorption; Cell Movement; Cells, Cultured; Cholesteatoma, Middle Ear; Chronic Disease; Ear Canal; Epithelium; Gene Expression; Humans; Immunohistochemistry; Keratins; Otitis Media; Phenotype; Skin

Several factors seem to contribute to the series of events in the pathogenesis of otitis media and cholesteatoma. Endotoxin is likely to be one of these factors, since it has been found in human middle ear effusions and since injection of this substance into the middle ear, in animal experiments, gave rise to prominent reactions. Provoking of epithelial cells in vitro with endotoxin led to distinct cell responses that might be associated with cholesteatoma formation. In this study the effect of endotoxin on serially cultured rat middle ear epithelium, rat meatal epidermis, and human keratinocytes was investigated. Endotoxin strongly stimulated the proliferation of middle ear epithelium and human keratinocytes and inhibited that of meatal epidermis. Furthermore, endotoxin affected the morphology of the three types of tissue. Rat middle ear epithelium revealed epithelial cell tracks with interconnecting bridge-like structures protruding above the culture plane, whereas rat meatal epidermis showed increased terminal differentiation expressing large areas of blister-like structures detaching from the culture dish. Cross-linked envelope analysis of human keratinocytes showed an increased terminal differentiation that was morphologically confirmed but was not confirmed by cytokeratin analysis. The results of this study support the hypothesis that endotoxin may play an important role in the pathogenesis of otitis media and cholesteatoma.

Topics: Animals; Antibodies, Monoclonal; Cell Movement; Cholesteatoma; Culture Techniques; Ear Ossicles; Ear, Middle; Endotoxins; Epidermis; Epithelium; Humans; Keratinocytes; Keratins; Otitis Media; Rats

The immunohistological characteristics of retraction pockets, cholesteatoma matrix and granulomatous tissue were compared in 14 samples from pediatric cholesteatoma. The junction between epidermis and the middle ear mucosa appeared as the most inflammatory area, displaying the characteristics of delayed type hypersensitivity. CD1 + Langerhans cells were observed in all epidermic areas, but expressed class II molecules only in the vicinity of polymorphonuclear infiltrates. Numerous mast cells and IgA producing cells were also observed, suggesting that defenses from the mucosal immune system are summoned and contribute to the pathogenesis of cholesteatoma.

Topics: Adolescent; Antigen-Presenting Cells; Child; Child, Preschool; Cholesteatoma; Ear, Middle; Female; Fluorescent Antibody Technique; Granuloma; Humans; Immunoglobulins; Keratins; Langerhans Cells; Male; Otitis Media; T-Lymphocyte Subsets

The expression of cytokeratins in the epithelium of the middle ear and external auditory meatus of the rat was studied on cryosections of ethylenediaminetetraacetic acid-decalcified specimens by use of a panel of monoclonal antibodies. The normal middle ear epithelium revealed a complex cytokeratin profile, including regional differences. The induction of sterile middle ear effusions resulted in increased cytokeratin expression. Infective effusions were accompanied by both quantitative and qualitative changes in the cytokeratin expression patterns. The differences observed between the cytokeratin profiles of external meatal skin and those of middle ear epithelium may form a useful tool for research into cholesteatoma development.

Topics: Animals; Antibodies, Monoclonal; Ear, Middle; Epithelial Cells; Hyperplasia; Hypertrophy; Immunohistochemistry; Keratins; Male; Mucous Membrane; Otitis Media; Rats; Rats, Inbred Strains

Amniotic fluid cellular content (AFCC), especially keratinized, non-nucleated squamous epithelial cells and lanugo hair, have been detected in the fetal and neonatal ear. We wished to evaluate the incidence, amount and histological findings of AFCC in the neonatal and early infant period. Temporal bones (n = 63) from children aged 10 min to 70 days were examined. AFCC was discovered in 39 of 43 bones in neonates. In children of 31-70 days of age, AFCC was present in 11 of 20 bones. A measurement of the amount of AFCC was performed; the maximum was 11.47 mm3 with a mean of 2.40 mm3. Histological findings varied from the mere presence of AFCC to the formation of epithelialized granulation tissue. This granulation tissue was found in 19 of the bones. Theoretical clinical implications are considered for middle ear adhesions, otitis media and cholesteatoma.

Topics: Amniotic Fluid; Child, Preschool; Cholesteatoma; Ear Diseases; Ear, Middle; Epidermal Cells; Epithelium; Foreign-Body Reaction; Granulation Tissue; Hair; Humans; Infant; Infant, Newborn; Keratins; Otitis Media; Skin; Temporal Bone; Tissue Adhesions

Keratin debris is a constant feature in middle-ear cholesteatoma. Keratin prepared from rat skin induced a foreign-body granuloma in the subcutaneous space in the rat. In vitro this granuloma produced high levels of bone-resorbing factors: prostaglandin E2, osteoclast-activating factor, and leucine aminopeptidase. In the in vivo study, keratin-induced granuloma in the rat middle ear caused partial resorption of the cochlear wall. Macrophages, fibroblasts, and osteoclastlike cells were found at bone-resorption areas. These cells appeared to be responsible for bone resorption through production of prostaglandin E2, osteoclast-activating factor, and proteases.

Topics: Animals; Bone Resorption; Carrageenan; Cholesteatoma; Dinoprostone; In Vitro Techniques; Keratins; Leucyl Aminopeptidase; Lymphokines; Male; Otitis Media; Prostaglandins E; Rats

In a study of the behavior of the keratin layer of the drumhead epidermis in some otologic conditions, a marker dye was applied to the tympanic membrane, and migration and/or desquamation of the dotted spots observed. Decelerated migration with desquamation in situ was observed during the quiescent phase of recurrent otitis externa, with or without myringitis. The same findings were observed in most cases of chronic secretory otitis media and its sequelas (retraction pockets, atrophic membranes, and scarring). The remnants of drumhead with keratotic and hyperplastic changes in the middle ear cleared themselves of the dye very slowly.

Topics: Adolescent; Adult; Aged; Child; Ear Diseases; Humans; Inflammation; Keratins; Middle Aged; Otitis Externa; Otitis Media; Skin; Tympanic Membrane

Topics: Adult; Aged; Autopsy; Child; Cholesteatoma; Chronic Disease; Connective Tissue; Ear, Middle; Epithelial Cells; Epithelium; Granulation Tissue; Humans; Hyperplasia; Hypertrophy; Keratins; Male; Metaplasia; Microtomy; Middle Aged; Mucous Membrane; Otitis Media; Staining and Labeling; Temporal Bone; Tympanic Membrane

Topics: Bone and Bones; Cholesteatoma; Chronic Disease; Ear Diseases; Ear Ossicles; Epithelium; Granulation Tissue; Humans; Keratins; Mucus; Otitis Media

Topics: Biopsy; Cell Differentiation; Ear, Middle; Epithelium; Humans; Infant; Keratins; Mucous Membrane; Otitis Media

Topics: Ear, External; Humans; Keratins; Male; Middle Aged; Otitis Media; Tympanic Membrane

Topics: Cholesteatoma; Humans; Keratins; Otitis Media; Surgical Procedures, Operative