In vitro study on immune response modifiers as novel medical treatment options for cholesteatoma.

OBJECTIVES: To investigate cytokine profile of cholesteatoma and to collect information about important intercellular signaling pathways by establishing two different cell culture models, to block important intercellular signaling pathways in cholesteatoma by applying immune system modifier drugs to develop alternative medical therapy options for cholesteatoma. METHODS: To observe the pathogenesis of cholesteatoma and to apply the immunomodulatory drugs, cholesteatoma tissue culture models were constituted with HEKa cells and cholesteatoma keratinocytes, which were obtained from 3 patients who underwent operations for cholesteatoma. Medicines including 5-fluorourasil, imiquimod, cyclosporine, and tacrolimus were applied on both cholesteatoma keratinocytes and HEKa cells. After 48 h of incubation, IL-1, IL-6, IL-8, IL-10, TNF-α, and Ki67 levels were measured to determine cell viability rates. RESULTS: In the cholesteatoma control group, IL-6 and TNF-α levels were found higher than in the HEKa control group. All repurposed drugs in the study demonstrated anti-inflammatory, anti-proliferative, and cytotoxic effects on cholesteatoma. Imiquimod and tacrolimus in particular are potential treatment prospects for cholesteatoma due to their strong anti-inflammatory and cytotoxic effects. CONCLUSION: Medical therapy options for cholesteatoma are still missing and surgery is not the ultimate solution. We have focused on intercellular inflammatory processes, which play significant roles in the pathogenesis of cholesteatoma in our paper. Inflammation and proliferation of cholesteatoma decreased after all repurposed drug applications in our study. Anti-inflammatory and anti-proliferative effects of tacrolimus and imiquimod was more significant than other drugs in the study. For this reason, tacrolimus and imiquimod should be examined in depth with in vivo studies in terms of efficacy and safety for medical treatment of cholesteatoma.

Hsa_circ_0074491 regulates the malignance of cholesteatoma keratinocytes by modulating the PI3K/Akt pathway by binding to miR-22-3p and miR-125a-5p: An observational study.

ABSTRACT: Cholesteatoma is a benign cystic lesion that can continue to grow like a tumor. Circular ribonucleic acid (RNA) hsa_circ_0074491 (circ_0074491) has been reported to be down-regulated in cholesteatoma tissues. However, the role and regulatory mechanism of circ_0074491 in the growth of cholesteatoma are unclear.The expression of circ_0074491, microRNA (miR)-22-3p, and miR-125a-5p in cholesteatoma tissues was detected by quantitative real-time polymerase chain reaction. The proliferation, cell cycle, apoptosis, migration, and invasion of cholesteatoma keratinocytes were evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, plate clone, flow cytometry, or transwell assays. Several protein levels were examined by western blotting. The targeting relationship between miR-22-3p or miR-125a-5p and circ_0074491 was verified via dual-luciferase reporter and RNA pull-down assays.We observed the downregulation of circ_0074491 in cholesteatoma tissues. Furthermore, circ_0074491 knockdown facilitated cell proliferation, migration, invasion, and repressed cell apoptosis in cholesteatoma keratinocytes. Circ_0074491 was verified as a decoy for miR-22-3p and miR-125a-5p in cholesteatoma keratinocytes. Both miR-22-3p and miR-125a-5p silencing reversed the impacts of circ_0074491 silencing on proliferation, apoptosis, migration, and invasion of cholesteatoma keratinocytes. Also, circ_0074491 knockdown activated the PI3K/Akt pathway in cholesteatoma keratinocytes via miR-22-3p and miR-125a-5p.Circ_0074491 played a suppressive role in cholesteatoma through inactivating the PI3K/Akt pathway via binding to miR-22-3p and miR-125a-5p, which provided a novel evidence for the involvement of circRNA in the development of cholesteatoma.

[Burow's solution treatment for external auditory canal and mastoid cavity cholesteatoma].

Burow's solution, 13% aluminum acetate dissolved in water developed as ear drops by German Dr. Karl August Burow in the mid-1800s, was confirmed by Mahoney (1980) and Thorp et al. (2000) to act on chronic suppurative otitis media without ototoxicity. We have found it satisfactory in treating otitis media and other intractable inflammation and fungal infection of the external auditory canal. We report its potent effect in 2 cases of external auditory canal cholesteatoma (EACC) and 1 of mastoid cavity cholesteatoma (MCC)-the first such report insofar as we knew. Case 1 of EACC involved an 8-year-old boy with cholesteatoma debris in a bony groove evidencing defective skin just posteroinferior to the left tympanic membrane. EACC was cured after a single Burow's solution instillation. Case 2 of EACC was a 31-year-old woman whose left ear canal was filled by a keratin mass with pus positive for methicillin-resistant staphylococcus aureus (MRSA) and granulation and experiencing otalgia and dysgeusia. Computed tomography (CT) showed that the bony anterior and posterosuperior wall of the external auditory canal had been destroyed. The EACC was removed and Burow's solution instilled once a week, effecting a cure about one month later, leaving the bony groove of the anterior and inferior wall intact and recovering gustatory sensation. Case 3 of MCC was a 47-year-old man undergoing right canal wall down tympanoplasty three times. One year after the last surgery, the mastoid cavity and posterior external canal wall was covered by a cholesteatoma matrix with granulation and pus. Instilling Burow's solution for 5 months resulted in a completely dry cholesteatoma-free cavity. All three subjects had intact tympanic membranes. Burow's solution alone proving effective against EACC and MCC suggests its great usefulness as first-choice conservative therapy. Although this usually cannot completely cure middle-ear and attic cholesteatoma, its effectiveness in cases of EACC and MCC should be recognized in infection and inflammation with cholesteatoma.

Clinical efficacy of 5-fluorouracil (5-FU) topical cream for treatment of cholesteatoma.

OBJECTIVE: In order to clarify clinical efficacy of commercially available 5-fluorouracil (5-FU) topical cream, a clinical study was conducted. METHODS: Two to three cubic millimetres of 5-FU topical cream (Kyowa, Roche) was applied on 50 cases of various types of cholesteatoma (50 patients) two to five times with the interval of 2 weeks, and its clinical efficacy was evaluated by the criteria we developed. RESULTS: In total, 59% of the cholesteatomas showed good effect, 29% of them showed fair effect, and the effect was poor in the remaining 12%. It was particularly effective in cholesteatomas in the EAC, attic cholesteatomas with an aerated mastoid, and in recurrent-type cholesteatomas. CONCLUSION: 5-FU topical cream appeared effective for the treatment of cholesteatomas.

The topical use of 5-fluorouracil in the ear in the management of cholesteatoma and excessive mucous secretion.

5-Fluorouracil was introduced in the early 1960s as a topical chemotherapeutic agent and has become increasingly accepted because of its efficacy, economy, and relative absence of side effects in treating many pre-cancerous conditions, certain benign and malignant tumors, and dermatoses. This paper addresses the use of topical 5-fluorouracil in the management of cholesteatoma and unwanted mucus-secreting cells. A retrospective study has suggested a very impressive efficacy of topical 5-fluorouracil in the management of cholesteatoma and, to a lesser degree, control of hyper-mucus-secreting cells at locations in the middle ear and mastoid. The use of topical 5-fluorouracil in the management of hyperkeratosis, cholesteatoma in the middle ear and mastoid, and in the external canal has been almost 100% effective with very few side effects. In the management of hyper-mucous-secreting cells, 5-fluorouracil has a success rate of approximately 50%.

Ciprofloxacin for the treatment of chronic ear disease.

The treatment of chronic ear disease is often difficult and frustrating. Patients typically present with a history of chronic, persistent otorrhea that has failed to respond to multiple topical and oral antibiotics. Organisms that are resistant to multiple antibiotics are common. Ciprofloxacin has been shown to be effective against a wide range of gram-negative and gram-positive organisms. To evaluate the role of ciprofloxacin in the treatment of chronic ear disease, 21 patients who failed routine therapy for chronic ear disease were prospectively treated with oral ciprofloxacin. Prior to therapy, all ear cultures grew Pseudomonas aeruginosa, Staphylococcus aureus or other gram-negative organisms. Ninety-five percent of patients completing therapy showed either improvement or cure. Only one patient failed to improve. Ciprofloxacin has been shown to be effective in the management of chronic ear disease.

Aminoglycoside antibiotics in the treatment of otologic infections.

Aminoglycoside antibiotics can be used successfully in the treatment of otologic infections caused by gram-negative bacteria. They can be particularly valuable, sometimes in combination with a beta-lactam antibiotic such as carbenicillin, in malignant external otitis, acute middle ear infections caused by gram-negative organisms, and central nervous system complications of cholesteatomas. On the basis of susceptibility tests and of the pharmacology of these drugs, we administer appropriate therapeutic doses of one of the following antibiotics: kanamycin, gentamicin, tobramycin, or amikacin. All of these drugs may be ototoxic and nephrotoxic.

First observations on treatment of cholesteatoma with (N-acetyl)cysteine.

In a series of cholesteatoma patients the benefit of local application of acetylcysteine was demonstrated. Chemical studies and fractional analyses were used to reveal the breakdown of keratin. Histochemically the influence of acetylcysteine on keratin is confirmed.

Topical applications of 5-fluorouracil in the medical treatment of cholesteatoma of the middle ear.

Recent studies have demonstrated that aural cholesteatoma features intense DNA turnover in the germinative layer of the matrix and an immune response in the wall of the cyst, which trigger a chain reaction. 5-Fluorouracil, an antimetabolitic agent that is also effective topically, was applied to the cyst wall in the initial cholesteatoma, in relapsing cholesteatoma (even transtympanically) and in large exudative cavities resulting from attico-antrectomy: in all cases, it inhibited the formation of keratin and reduced the mucous hypersecretion. These results have persisted even after 12 months of suspending treatment.

Biofilm formation by otopathogenic strains of Pseudomonas aeruginosa is not consistently inhibited by ethylenediaminetetraacetic acid.

HYPOTHESIS: Biofilm formation in otopathogenic of Pseudomonas aeruginosa (OPPA) strains is inhibited by ethylenediaminetetraacetic acid (EDTA). BACKGROUND: EDTA, a widely used chelating agent, has been shown to inhibit biofilm formation in a number of bacteria. Because EDTA may be a well-tolerated reagent to inhibit biofilm formation in cases of suppurative otitis media, we asked if it might be effective in all OPPA strains isolated from chronically infected cholesteatomas. METHODS: OPPA strains were isolated from patients with infected cholesteatomas. These strains were grown into log phase then were placed in minimal media with varying concentrations of EDTA and incubated for varying periods. Biofilm production was measured colorimetrically by staining with crystal violet. RESULTS: Without added EDTA, most otopathogenic PA exhibited a distinct, but varying, time course of biofilm formation and dissolution with peak production at 12 to 18 hours. Addition of 1 mM EDTA resulted in a delay in the time to peak biofilm formation for most strains, although the amount of biofilm was not decreased. In contrast, some strains showed greater biofilm production with 1 mM EDTA compared with the untreated bacteria. Addition of 10 mM EDTA resulted in a similar effect. Some strains increased biofilm production over controls. Moreover, EDTA inhibited planktonic growth of all OPPA strains at the concentrations studied. CONCLUSION: Our hypothesis was disproven: EDTA tends to delay biofilm development, although it consistently inhibits planktonic growth. Because EDTA does not cause suppression of biofilm production in all isolates of OPPA, usefulness as an antimicrobial is questioned.

Suppressive activity of vitamin D3 on matrix metalloproteinase production from cholesteatoma keratinocytes in vitro.

There is much evidence that degradation of the extracellular matrix is essential for the development of cholesteatomas and that this is induced by activation of matrix metalloproteinases (MMPs). Vitamin D3 (VD3) has several well-recognised biological activities, including suppression of MMP production. The present study, therefore, was undertaken to examine whether VD3 could suppress MMP production from cholesteatoma keratinocytes in vitro. Keratinocytes (2.5 x 10(5) cells/mL) induced from cholesteatoma tissue specimens were cultured with various concentrations of VD3. After one hour, lipopolysaccharide was added to the cell cultures at 100 mug/mL. The culture supernatants were then collected and assayed for MMP-1 and MMP-3 by ELISA. We also used ELISA to measure the levels of both TIMP (tissue inhibitor of metalloproteinase)-1 and TIMP-2 in culture supernatants. Addition of VD3 into keratinocyte cultures caused the suppression of MMP and TIMP production, which was increased by LPS stimulation. This was dose-dependent. The present results showing the suppressive activity of VD3 on the production of MMPs, which are responsible for tissue remodeling, strongly suggest that VD3 would be a good candidate for an agent in the medical treatment of, or prophylaxis for, cholesteatomas.

[A new medical therapeutic approach to cholesteatoma. 5-fluorouracil in local treatment]. / Une nouvelle approche thérapeutique médicale du cholestéatome. Le 5-fluorouracil en traitement local.

The use of applied 5-Fluorouracil is effective in the treatment of cholesteatoma. The authors described the limits of this technique.

Cholesteatoma of the renal pelvis: a case with long-term followup.

We report a case of cholesteatoma of the renal pelvis with a 56-year followup. Quiescence of the disease with remission of symptoms, stabilization of the lesion and absence of further anatomical change was related temporally to treatment with vitamin A. The disease in our patient was idiopathic and no causative or predisposing factors were demonstrated.

[Antimicrobial effectiveness of azlocillin in cholesteatoma in parenteral administration]. / Die antimikrobielle Wirksamkeit von Azlocillin in Cholesteatomen bei parenteraler Applikation.

Azlocillin has a broad spectrum particularly against pseudomonas and should therefore be useful for preoperative parenteral treatment of infected cholesteatomas in those cases which cannot be controlled by local therapy. To determine the efficacy in treatment of cholesteatoma the concentration of azlocillin in separate tissue fractions of cholesteatoma matrix, cholesteatoma debris and granulation tissue surrounding cholesteatoma was determined. In 9 cases 2 g azlocillin was injected and in 5 cases 4 g. The concentration drop between the different tissue fractions was equal in all cases. However, as clinical experience has shown, differing high levels of azlocillin were found in comparable tissue fractions of the 14 cholesteatomas. In some of the cases the minimal antibacterial activity was not attained in the cholesteatoma fractions.

The use of fluoroquinolones in chronic otitis suppurativa.

The present review covers fluoroquinolone usage in chronic otitis suppurativa (COS) in case of chronic otitis media, cholesteatoma, radical mastoid cavity infection and chronic or relapsing otitis externa. A total of six publications were included in the final evaluation. Enoxacin was effective in 35%; ciprofloxacin (five publications) was used in 82 patients with 67% effectivity in otitis externa and otitis media and 61% effectivity in radical mastoid cavity infection. No serious adverse reactions were reported. The promising efficacy of otitis media and otitis externa and safety profile needs further confirmation in double blind prospective clinical studies, that will provide a firm basis for changing the current treatment schedules of COS.