The Possible Role of Sclerostin in the Pathogenesis of Tympanosclerosis.

OBJECTIVE: The aim of this study was to investigate sclerostin (SOST) expression in a rat model of experimental tympanosclerosis (TS) and its possible role in the formation of TS. MATERIALS AND METHODS: Thirty-four SD rats were randomly divided into 2 groups: experimental group (n = 17) and normal group (n = 17). The left tympanic cavities in the experimental group were inoculated with methicillin-resistant Staphylococcus aureus. The changes of tympanic membranes were examined and recorded under otoendoscope. Haematoxylin-eosin staining was adopted to detect the morphological changes in the tympanic membrane and middle ear mucosa. Immunohistochemistry and Western blot analysis were used to observe the expression of SOST, Wnt3a, ß-catenin, and P-ERK1/2. RESULTS: In the experimental group, sclerotic lesions were observed in 54.5% ears in the end of 6 weeks. Morphological changes such as mucosa incrassation, inflammatory cells infiltration, fibrous tissue proliferation, and interstitial tissue incrassation prominently appeared in the tympanic membrane and middle ear mucosa. SOST protein was mainly distributed in the cytoplasm of epithelial cells and gland cells, the expression of which increased significantly in the calcified experimental ears. In addition, expression levels of Wnt3a, ß-catenin, and P-ERK1/2 increased significantly in the calcified group too. CONCLUSION: The upregulated expression level of SOST may be involved in the formation of TS, first, through the pro-phosphorylation of ERK1/2 in the inflammatory stage, and then through the enhancement of Wnt3a in the osteogenic stage.

Comparison of the Aditus ad Antrum Diameter on CT between Patients with Chronic Otitis Media and Healthy Individuals.

BACKGROUND: The middle ear cavity is ventilated through the aditus ad antrum. Aditus blockage contributes to the pathology of otitis media. OBJECTIVE: To determine the normal values of the aditus ad antrum diameter on computed tomography and to investigate its relationship with chronic otitis media and related pathologies (tympanosclerosis and myringosclerosis). METHODS: The temporal CT images of 162 individuals were evaluated retrospectively. In the axial sections, the inner diameter of the aditus was measured at the narrowest point at the cortex. The differences in diameter were compared between diseased and healthy ears. RESULTS: In healthy individuals, the diameter was narrower in women. There was no difference between the right and left ears in healthy subjects. No correlation was found between age and diameter. In male patients with myringosclerosis, the diameter was slightly narrower on both sides but more marked on the left. In female patients with myringosclerosis, the diameter in both ears was slightly narrower. In cases of otitis media and tympanosclerosis, the diameter was less than that of healthy individuals, despite the lack of statistically significant result in all cases. CONCLUSION: The aditus ad antrum was narrower in diseased ears, indicating that a blocked aditus may contribute to the development of otitis media, as well as mucosal diseases.

Is there any effect of montelukast on prevention of myringosclerosis after myringotomy in a rat model?

OBJECTIVES: In this study, our aim was to identify the possible effects of montelukast sodium (ML) on the prevention of experimentally induced myringosclerosis. MATERIALS AND METHODS: Twenty-eight female Wistar albino rats were used and they were divided into four groups randomly. Tympanic membranes (TM) of all animals were perforated and then group 1 received no treatment (control group), group 2 was treated with a topical saline solution, group 3 received topically ML and group 4 received orally ML. On the 15th day, all animals were euthanized. Tympanic membranes were evaluated otomicroscopically and histopathologically. RESULTS: The histopathological findings, compared against a control and saline groups, showed the topically and orally ML groups had statistically significant differences of degree of myringosclerosis (p < 0.002) and median thickness of the TMs (p < 0.001). Suppression of inflammation was statistically significant only in the oral ML treatment group (p < 0.002). CONCLUSION: Oral and topically administration of ML reduced myringosclerosis formation in myringotomies rats.

Histopathology of tympanic membranes from patients with ventilation tubes.

OBJECTIVE: To evaluate the histopathologic changes in tympanic membranes (TMs) with ventilation tubes (VTs). METHODS: In this retrospective human temporal bone study our overall study group included 4 subgroups of TMs from deceased donors as follows: 24 with a history of VT insertion for chronic otitis media with effusion (COME-VT); 5 with a history of VT insertion for Meniere's disease (MD-VT); 33 without a history of VT insertion for chronic otitis media with effusion (COME); and 14 without a history of VT insertion for Meniere's disease (MD). We classified the extent of migration of the outer keratinized squamous epithelium onto the inner surface of TM perforations and noted the presence and location of tympanosclerosis, of atrophy, of perforation, and/or of cholesteatoma formation. RESULTS: Tympanosclerosis occurred in 14/24 TMs in the COME-VT subgroup; 2/5, MD-VT; 7/33, COME; and 0/14, MD. The VT insertion site was mostly in the anteroinferior (63%) quadrant of the TM; tympanosclerosis occurred more frequently in the posteroinferior (42%) and posterosuperior (33%) quadrants. We found no significant correlation between the location of tympanosclerosis and the VT insertion site (P>0.05). Atrophy occurred in 7/24 TMs in the COME-VT subgroup; 3/5, MD-VT; 8/33, COME; and 2/14, MD. We found no significant correlation between the location of atrophy and the VT insertion site; however, atrophy was located mostly in the anteroinferior quadrant (one of the most common VT insertion sites) of the TM. Regarding the ingrowth of keratinized epithelium, the mucocutanous junction was detected at any point at the inner surface of the TM in 50% of the specimens. We observed intratympanic cholesteatoma formation in 2/24 TMs in the COME-VT subgroup. CONCLUSION: TM changes due to VT insertion are more common than previously realized. Meticulous otomicroscopic evaluation of the TM is necessary during tympanomastoidectomies in order to prevent the intratympanic inclusion pearls and squamous epithelial ingrowth to prevent any further cholesteatoma formation.

The effect of melatonin and vitamin C treatment on the experimentally induced tympanosclerosis: study in rats / Efeito do tratamento com melatonina e vitamina C na timpanoesclerose induzida experimentalmente: estudo em ratos

Abstract Introduction: The ethiopathogenesis of tympanosclerosis has not been completely under- stood yet. Recent studies have shown that free oxygen radicals are important in the formation of tympanosclerosis. Melatonin and Vitamin C are known to be a powerful antioxidant, interacts directly with Reactive Oxygen Species and controls free radical-mediated tissue damage. Objective: To demonstrate the possible preventative effects of melatonin and Vitamin C on tympanosclerosis in rats by using histopathology and determination of total antioxidant status total antioxidant status. Methods: Standard myringotomy and standard injury were performed in the middle ear of 24 rats. The animals were divided into three groups: Group 1 received melatonin, Group 2 received vitamin C, and Group 3 received saline solution. Results: The mean values of total antioxidant status were similar in the all study groups before the treatment period. The mean values of total antioxidant status were significantly higher in the melatonin and vitamin C groups compared to control group but vitamin C with melatonin groups were similar after the treatment period (p < 0.001). Minimum and maximum wall thicknesses were lower in the melatonin and vitamin C groups compared to the control group but the differences were insignificant. Conclusion: Melatonin increases total antioxidant status level and might have some effect on tympanosclerosis that develops after myringotomy.

Resumo Introdução: A etiopatogênese da timpanoesclerose (TE) não foi ainda totalmente esclarecida. Estudos recentes têm demonstrado que os radicais livres de oxigênio são importantes na formação de TE. Melatonina e vitamina C são conhecidas por serem poderosos antioxidantes, interagir diretamente com espécies reativas de oxigênio (ROS) e controlar danos em tecidos mediados por radicais livres. Objetivo: Demonstrar os possíveis efeitos preventivos da melatonina e da vitamina C na TE em ratos com histopatologia e determinação da capacidade antioxidante total (CAT). Método: Miringotomias padronizadas foram feitas na orelha média de 24 ratos. Os animais foram divididos em três grupos: o Grupo 1 recebeu melatonina, o Grupo 2 vitamina C e o grupo 3 solução salina. Resultados: Os valores médios de CAT foram semelhantes em todos os grupos de estudo antes do período de tratamento. Os valores médios de CAT foram significativamente maiores nos grupos que receberam melatonina e vitamina C em comparação com o grupo de controle, mas os grupos vitamina C e melatonina foram semelhantes após o período de tratamento (p < 0,001). As espessuras mínimas e máximas de parede foram menores nos grupos melatonina e vitamina C, em comparação com o grupo controle, mas as diferenças não foram significativas. Conclusão: A melatonina aumenta os níveis de CAT e pode ter algum efeito sobre a TE que se desenvolve após a miringotomia.

The effects of fibroblast growth factor-2 delivered via a Gelfoam patch on the regeneration of myringosclerotic traumatic eardrum perforations lying close to the malleus.

OBJECTIVE: We evaluated the effects of fibroblast growth factor-2 (FGF-2) delivered via a Gelfoam patch on the regeneration of myringosclerotic traumatic tympanic membrane perforations (TMPs) lying close to the malleus. STUDY DESIGN: A prospective, randomized, controlled clinical study. SETTING: A university-affiliated teaching hospital. SUBJECTS AND METHODS: We prospectively analyzed, in a randomized manner, the outcomes of treatment for traumatic TMPs constituting >25% of the tympanic membrane. The closure rates, closure times, and otorrhea rates were compared among patients treated via FGF-2-containing Gelfoam patches, Gelfoam patches alone, and observation only. RESULTS: We analyzed data from 138 patients. The perforation closure rates in the FGF-2 plus Gelfoam patch, Gelfoam patch, and observation alone groups were 97.9, 89.8, and 70.7%, respectively. Both the FGF-2 plus Gelfoam and Gelfoam alone groups exhibited significantly higher closure rates than the observational group (both p<0.05).The mean closure times were 15.7±5.1, 24.8±4.9, and 35.7±9.2days in the FGF-2 plus Gelfoam patch, Gelfoam patch alone, and observation alone groups, respectively. The FGF-2 plus Gelfoam patch group exhibited a significantly shorter closure time than the Gelfoam patch alone and observation alone groups (p<0.05). The incidences of purulent otorrhea were 14.6, 6.1, and 4.9% in the FGF-2 plus Gelfoam patch, Gelfoam patch alone, and observation alone groups, respectively. Surprisingly, 7 of 7 (100.0%) perforations associated with purulent otorrhea completely closed in the FGF-2 plus Gelfoam patch group; however, no such perforation healed in either the Gelfoam alone or observation alone group. CONCLUSIONS: FGF-2 plus Gelfoam patching significantly shortened the closure time compared to observation and Gelfoam patching alone, and it significantly improved the closure rate (compared to observation alone) of myringosclerotic perforations lying close the malleus. FGF-2 plus Gelfoam patching is a valuable, minimally invasive alternative treatment that may be readily applied to outpatient settings.

Does calcemia influence the onset of myringosclerosis after myringotomy with the insertion of ventilation tubes?

INTRODUCTION: Myringosclerosis is one of the most frequent late complications of the insertion of ventilation tubes, and its aetiopathogenesis remains unknown. The calcification that occurs in the formation of myringosclerosis plaque raises the hypothesis of the presence of a calcium metabolism disorder. The objective is to determine whether calcemia contributes to the development of myringoscelerosis after insertion of ventilation tubes. MATERIAL AND METHODS: A longitudinal, prospective, analytical cohort study was conducted in patients undergoing myringotomy with the insertion of ventilation tubes due to otitis media with effusion. Calcemia was evaluated pre-operatively and in the follow-up the appearance of myringosclerosis and the percentage of the tympanum involved were evaluated. RESULTS: The study included 156 patients (297 ears), with calcemia values ranging from 7.6 to 10.2mg/dl. Myringosclerosis was identified in 35.4% of the operated ears. No relationship was found between the appearance of myringosclerosis and calcemia (p=.596). It was found, however, that the greater the percentage of the tympanum affected by myringosclerosis, the lower the calcemia values (p=.014). CONCLUSION: The population studied had calcemia values within the normal range, which allows us to infer that no changes in calcium metabolism are required for the development of myringosclerosis. Moreover, unlike in previous studies, higher calcium levels are not associated with more myringosclerosis. Calcemia did not influence the appearance of myringosclerosis after myringotomy with the insertion of ventilation tubes.

The effect of melatonin and vitamin C treatment on the experimentally induced tympanosclerosis: study in rats.

INTRODUCTION: The ethiopathogenesis of tympanosclerosis has not been completely under- stood yet. Recent studies have shown that free oxygen radicals are important in the formation of tympanosclerosis. Melatonin and Vitamin C are known to be a powerful antioxidant, interacts directly with Reactive Oxygen Species and controls free radical-mediated tissue damage. OBJECTIVE: To demonstrate the possible preventative effects of melatonin and Vitamin C on tympanosclerosis in rats by using histopathology and determination of total antioxidant status total antioxidant status. METHODS: Standard myringotomy and standard injury were performed in the middle ear of 24 rats. The animals were divided into three groups: Group 1 received melatonin, Group 2 received vitamin C, and Group 3 received saline solution. RESULTS: The mean values of total antioxidant status were similar in the all study groups before the treatment period. The mean values of total antioxidant status were significantly higher in the melatonin and vitamin C groups compared to control group but vitamin C with melatonin groups were similar after the treatment period (p<0.001). Minimum and maximum wall thicknesses were lower in the melatonin and vitamin C groups compared to the control group but the differences were insignificant. CONCLUSION: Melatonin increases total antioxidant status level and might have some effect on tympanosclerosis that develops after myringotomy.

Ectopic Mineralization and Conductive Hearing Loss in Enpp1asj Mutant Mice, a New Model for Otitis Media and Tympanosclerosis.

Otitis media (OM), inflammation of the middle ear, is a common cause of hearing loss in children and in patients with many different syndromic diseases. Studies of the human population and mouse models have revealed that OM is a multifactorial disease with many environmental and genetic contributing factors. Here, we report on otitis media-related hearing loss in asj (ages with stiffened joints) mutant mice, which bear a point mutation in the Enpp1 gene. Auditory-evoked brainstem response (ABR) measurements revealed that around 90% of the mutant mice (Enpp1asj/asj) tested had moderate to severe hearing impairment in at least one ear. The ABR thresholds were variable and generally elevated with age. We found otitis media with effusion (OME) in all of the hearing-impaired Enpp1asj/asj mice by anatomic and histological examinations. The volume and inflammatory cell content of the effusion varied among the asj mutant mice, but all mutants exhibited a thickened middle ear epithelium with fibrous polyps and more mucin-secreting goblet cells than controls. Other abnormalities observed in the Enpp1 mutant mice include over-ossification at the round window ridge, thickened and over-calcified stapedial artery, fusion of malleus and incus, and white patches on the inside of tympanic membrane, some of which are typical symptoms of tympanosclerosis. An excessive yellow discharge was detected in the outer ear canal of older asj mutant mice, with 100% penetrance by 5 months of age, and contributes to the progressive nature of the hearing loss. This is the first report of hearing loss and ear pathology associated with an Enpp1 mutation in mice. The Enpp1asj mutant mouse provides a new animal model for studying tympanosclerotic otitis and otitis media with effusion, and also provides a specific model for the hearing loss recently reported to be associated with human ENPP1 mutations causing generalized arterial calcification of infancy and hypophosphatemic rickets.

The effect of Nigella sativa oil on prevention of myringosclerosis in a Guinea pig model.

OBJECTIVES: In this study, our aim was to identify the possible effects of Nigella sativa L. (NS) [blackcumin] seed oil on the prevention of experimentally induced myringosclerosis (MS). MATERIALS AND METHODS: Fourteen Guinea pigs were used and they were divided into three groups. Tympanic membranes (TM) of all animals were perforated and then group I was treated with saline soaked gel foams as a control group, group II was treated with 0.5 ml NS oil soaked gel foams at 0, 24 and 48 h and group III was treated with 5 ml NS oil orally at 0, 24, 48, 72 and 120 h. After 15 days, all animals were euthanized. Tympanic membranes were evaluated macroscopically and histopathologically. RESULTS: Groups I showed extensive myringosclerosis in contrast to those of Groups II and III which had significantly less changes (p < 0.05). The fibrosis and inflammation in the lamina propria of the tympanic membranes of Groups I was found to be significantly more pronounced (p < 0.05). The tympanic membranes were found to be significantly thinner in Groups II and III when compared with Groups I (p < 0.05). CONCLUSIONS: The results of this study suggested that topical or oral administration of NS oil supressed the inflammation and fibroblastic activity in the lamina propria of the myringotomized TMs of the Guinea pigs. For providing further evidence to use plant extracts as antioxidant and antiinflammatory therapy after myringotomy or ventilation tube insertion, further clinical studies with larger population will be essential.

The impact of topical and systemic enoxaparin sodium use on traumatic tympanic membrane perforation and myringosclerosis.

The objective of this study is to investigate the effect of topical and systemic enoxaparin sodium on the healing pattern of experimentally induced tympanic membrane perforation and formation of myringosclerosis. A total of 24 Wistar-Albino strain rats were included in the study. Standard myringotomies were performed on each rat. In the first group, isotonic serum physiologic was dropped on external ear canal (control group). Topical enoxaparin was dropped on external ear canal and daily topical doses of enoxaparin were dropped on external ear canal of the rats for 14 days (topical treatment group). Third group received subcutaneous injections of enoxaparin for 14 days (systemic treatment group). Five micrometer thick sections of the bullae of the rats were stained with H&E. Inflammation, edema and sclerotic lesions and neovascularization observed in the lamina propria layer of the tympanic membrane, and total thickness of the tympanic membrane were evaluated. In intergroup comparisons, significant difference in the distribution pattern of severity of inflammation in all three groups was not observed (p = 0.784, p > 0.05). Total TM thickness differed among all three groups (p = 0.028, p < 0.05). A statistically significant difference was observed between the systemic enoxaparin and the control groups (p = 0.022, p < 0.05). A statistically significant difference was observed between the topical enoxaparin and the control groups (p = 0.037, p < 0.05). However, comparison between the topical and systemic treatment groups could not reveal any statistically significant intergroup difference (p = 0.682, p > 0.05). A significant difference was not observed among three groups as for the distribution of myringosclerotic plaques, severity of edema and neovascularization in the lamina propria (p = 0.539, p > 0.05), (p = 0.063, p > 0.05), (p = 0.152, p > 0.05). Topical and systemic enoxaparin treatment did not prevent formation of sclerotic plaques; however, it decreased TM thickness significantly in comparison with the control group.

Role of Chlamydia pneumoniae and Helicobacteria pylori in the development of tympanosclerosis.

The etiology of tympanosclerosis (TS) is not known, but TS commonly develops secondary to acute and chronic otitis media (COM). Since calcification process in TS resembles that of atherosclerosis (AS), pathogens that are related to pathogenesis of AS may be involved in development of TS. This prospective and controlled study, performed at a tertiary referral center, investigated a possible relationship between the presence of Chlamydia (C.) pneumoniae and Helicobacter (H.) pylori and the development of a tympanosclerotic plaque. The presence of C. pneumoniae was examined in the surgical specimens of 62 patients (29 females and 33 males; age range 10-70 years, mean age 30.8 ± 13.3 years), including 30 patients with TS, 14 patients with cholesteatoma, and 18 patients with chronic suppurative otitis media (CSOM). The presence of H. pylori was examined in the surgical specimens of 88 patients (41 females and 47 males; age range 6-70 years, mean age 32.5 ± 14.8 years), including 35 patients with TS, 22 patients with cholesteatoma, 20 patients with CSOM, and 11 patients with otosclerosis. Tympanosclerotic plaques and control specimens from the cholesteatoma, polypoid mucosa, or mucosal portion of the perforations and stapes supra structure were examined for the presence of H. pylori and/or C. pneumoniae using real-time polymerase chain reaction analysis. The analysis demonstrated that specimens from the tympanosclerotic plaques and the other types of COM were all negative for C. pneumoniae and H. pylori. An association between C. pneumoniae or H. pylori infection and the development of TS or other types of COM could not be established.

The Impact of Systemic and Local Administration of Ascorbic Acid on Traumatic Perforation of Tympanic Membrane and Myringosclerosis.

OBJECTIVE: In the present study, tympanic membranes (TM) harvested from myringotomized rats were analyzed histopathologically to compare the systemic and local effects of ascorbic acid on the development of myringosclerosis. MATERIALS AND METHODS: Forty male Wistar-Albino rats weighing between 350-400 g were included in this study. Under otomicroscopic examination, a standard 2-mm myringotomy incision was made on the posteroinferior quadrant of the TM of both ears. Rats were randomized into five groups as control, topical ascorbic acid 50 mg/kg, systemic ascorbic acid 50, 100, and 200 mg/kg groups, each group containing eight rats. On the 15th day of the study, the rats were decapitated, and bullas of the rats were extracted. Sections were stained with hematoxylin-eosin and examined through light microscopy. Inflammation, distribution width of plaques, edema, and neovascularization were observed on the lamina propria. Thickness of the TM was evaluated under the microscope and scored semiquantitatively. RESULTS: When intergroup comparisons of parameters related to total TM thickness were performed, differences between the control group and topical AA (ascorbic acid) or systemic treatment groups were found to be statistically significant (p<0.005). A statistically significant difference was detected among control, topical and systemic 200 mg/kg ascorbic acid groups for the edematous lamina propria (p=0.003 and p<0.05, respectively). CONCLUSION: For the total TM thickness, systemic and topical ascorbic acid use was effective when compared with the control group. It has been concluded that systemic use of higher doses of (200 mg/kg) ascorbic acid is beneficial in the resolution of the edematous lamina propria.

Increased carotid artery intima-media thickness in patients with tympanosclerosis: Common risk factors with atherosclerosis?

We investigated the relationship between tympanosclerosis, known atherosclerotic risk factors, and the intima-media thickness of bilateral carotid arteries using ultrasonography. A total of 122 patients admitted to our clinic with chronic otitis media between 2005 and 2010 were included in the study. The study group consisted of 61 patients with tympanosclerosis; the control group comprised 61 patients without tympanosclerosis. Internal carotid artery intima-media thickness (CAIMT), total cholesterol, triglyceride, low- and high-density lipoprotein cholesterol, C-reactive protein, and homocysteine levels were measured in all patients. Homocysteine, low-density lipoprotein, total cholesterol, and triglyceride levels in the study group were higher compared with those of the control group (p < 0.05). Right and left CAIMT was greater in the study group versus the control group (p ≤ 0.001). In conclusion, atherosclerosis and tympanosclerosis were associated with identical risk factors; in the tympanosclerosis group, CAIMT was increased significantly.

Does systemic clarithromycin therapy have an inhibitory effect on tympanosclerosis? An experimental animal study.

OBJECTIVE: To demonstrate the inhibitory effects of clarithromycin on in vitro tympanosclerosis. METHOD: Twenty-eight rats were divided into three groups: a clarithromycin group, a non-clarithromycin group and a negative control group. Those in the first two groups were injected with Streptococcus pneumoniae following a myringotomy, and tympanosclerosis was experimentally induced. Oral clarithromycin therapy was administered in the clarithromycin group. The other groups received no medical treatment. RESULTS: All eardrums in the clarithromycin and non-clarithromycin groups developed myringosclerosis, but there was only one eardrum, in the clarithromycin group, with very severe myringosclerosis. In the clarithromycin group, 11 ears showed no inflammation and there were no ears with severe inflammation. In the non-clarithromycin group, there were 11 ears with severe inflammation. The mean eardrum thickness in the clarithromycin group was 20.93 µm and in the non-clarithromycin group it was 42.71 µm. CONCLUSION: Acute otitis media and myringotomies induced tympanosclerosis, but clarithromycin reduced the severity of tympanosclerosis.

The effect of lycopene on experimental myringosclerosis.

OBJECTIVES: The aim of this study was to investigate the effect of lycopene on myringosclerosis development using histopathological and immunohistochemical analyses. METHODS: Fifty-six intact tympanic membranes of 28 guinea pigs were included in the study. Subjects were randomly divided into four groups (n=7/group). Group I (control group) did not receive any treatment after myringotomy. Group II (lycopene treatment after myringotomy) received oral lycopene (once daily at the same time, 10mg/kg, dissolved in water, administered with a catheter). The treatment lasted seven days. Group III (lycopene treatment before and after myringotomy), received lycopene treatment (same dose and route of administration) for seven days. Myringotomy was performed on day 8, and lycopene treatment was initiated immediately, and continued for seven days (same dose and route of administration). Group IV (lycopene treatment before myringotomy) received lycopene treatment one week before myringotomy using the same method and dose for seven days. Myringotomy was performed on day 8. Lycopene was not administered after myringotomy. Fourteen days after myringotomy, myringosclerosis was evaluated automicroscopically and scored. Following decapitation, bulla were removed and immersed in a 10% formaldehyde solution. Sections were cut for histopathological and immunohistochemical examination, and thickness, sclerosis, inflammation, and collagen-IV accumulation were scored semi-quantitatively. RESULTS: In the present study, the level of myringosclerosis was significantly lower in lycopene-treated groups compared to the control group (p<0.05). In addition, thickness, inflammation, sclerosis, and collagen-IV accumulation were significantly lower in the lycopene-treated groups compared to the control group (p<0.05). The timing of lycopene administration - i.e. before and/or after surgery - did not cause any difference with respect to myringosclerosis development. CONCLUSION: Lycopene, a strong antioxidant, may represent a good alternative treatment to prevent the development of myringosclerosis.

Effect of Enoxaparin Sodium on Experimentally-Induced Myringosclerosis in Rats.

OBJECTIVE: To evaluate the effectiveness of enoxaparin sodium (ES) on experimentally-induced myringosclerosis in rats. MATERIALS AND METHODS: Twenty Wistar albino-type rats weighing up to 250-300 g each were randomized into four groups containing five rats each and were then bilaterally myringotomized. The control group (n=5) received intratympanic serum physiologic injections, whereas ES2 (n=5), ES4 (n=5), and ES6 groups (n=5) received intratympanic ES of 2000 IU, 4000 IU, and 6000 IU, respectively, for 10 days after myringotomy. Rats were sacrificed at 60 days after intratympanic application and were then prepared for histopathologic evaluation. RESULTS: As for tympanic membrane hyaline degeneration, there were statistically significant differences among the control, ES2, ES4, and ES6 groups (p<0.05). As for fibrosis formation on tympanic membranes, a statistically significant difference was observed among the control and study groups; however, although not statistically significant, the formation of fibrosis was slowed down in the ES2 and ES4 groups compared with the control group. The control and study groups did not show any significant difference for calcification, hyperemia, and tympanic membrane thickening (p>0.05). CONCLUSION: Although our study and control groups comprised limited number of animals, and only one parameter demonstrated a statistically significant difference between the groups, ES may have an ameliorating effect on myringosclerosis induced by myringotomy in the tympanic membranes of rats. ES proved to be effective in the prevention of hyaline disc formation. Further studies should be conducted for better understanding of the effects of low-molecular-weight heparin (LMWH) (i.e., enoxaparin) on myringosclerosis.

Surgery of isolated malleus fixation due to tympanosclerosis.

The aim of the present study was to describe our surgical approach for isolated malleus fixation in patients with tympanosclerosis and to analyze the postoperative results. A total of 30 patients presented with isolated malleus fixation were operated. The fixation was reached via canalplasty. Fixated areas were cleaned without damaging the ossicle. Pre- and postoperative audiometric results were evaluated for each patient. Improvement of the pure-tone average (PTA) by at least 10 dB and an air-bone gap (ABG) of less than 20 dB after 12 months of follow-up was accepted to indicate success. The recovery of the postoperative PTA and ABG measurements was significant. Pre- and postoperative PTA was 48.00 ± 11.86 and 24.90 ± 12.45 dB, respectively (p < 0.001). According to PTA measurements, 40-50 dB recovery was achieved in four (13.3 %) patients, 31-40 dB in six (20 %) patients, 21-30 dB in ten (33.3 %) patients, and 11-20 dB in five (16.6 %) patients, with a total success rate of 25/30 (83.2 %). Pre- and postoperative ABG levels were 38.95 ± 9.92 and 16.10 ± 7.79 dB (p < 0.001), respectively. The ABG level was between 0 and 10 dB for 8 (26.6 %) patients, and 11-20 dB for 16 (53.3 %), with a total success rate of 24/30 (80 %). In cases of isolated malleus fixation with tympanosclerosis, performing a canalplasty to clean the sclerotic plaques without damaging the normal anatomy of the ossicle system using a diamond burr is a safe surgical option that provides significant recovery in hearing levels.

Effect of caffeic acid phenethyl ester on myringosclerosis development in the tympanic membrane of rat.

Myringosclerosis is hyalinization and calcification of certain areas of the tympanic membrane, especially the fibrous lamina propria layer and appears as white sclerotic lesions. Ventilation tube insertion is one of the most performed operations in the pediatric otorhinolaryngology practice to treat chronic otitis media with effusion. Myringosclerosis is a very common sequela of ventilation tube insertion. In this experimental study, our aim was to show the histopathological effects of caffeic acid phenethyl ester on myringosclerosis development in rat tympanic membrane after myringotomy. The rats were randomly categorized into four experimental groups including the comparison group (n = 4), non-treated group (n = 7), the saline (control) group (n = 7), the caffeic acid phenethyl ester group (n = 7). Non-treated group did not receive any treatment for 15 days. Saline (2.5 mL/kg, intraperitoneal) was administered to the third group once a day for 15 days. Fourth group received caffeic acid phenethyl ester intraperitoneally once a day at a dose of 10 µmol/kg for 15 days. Myringotomy was performed on the right tympanic membrane of all rats except comparison group using a sterile pick with the help of an operating microscope. Histopathological examination of myringosclerosis formation was done by a pathologist under light microscope. In histopathological analysis of groups, the severity of inflammation was milder in caffeic acid phenethyl ester group compared to non-treated and saline groups (p < 0.05). There was less myringosclerotic plaques in caffeic acid phenethyl ester group than in non-treated and saline groups (p < 0.05). TM thickness measurements were very close to each other in non-treated and saline groups. The tympanic membrane thickness of caffeic acid phenethyl ester group was much thinner than the other two groups (p < 0.05). Caffeic acid phenethyl ester decreases inflammation severity and the formation of myringosclerotic plaques. These two effects resulted in thinner tympanic membranes of rats which were treated with caffeic acid phenethyl ester. As a result, caffeic acid phenethyl ester has potential preventive effects on myringosclerosis development after myringotomy and ventilation tube insertion.

Combining use of captopril and losartan attenuates the progress of Streptococcus pneumoniae-induced tympanosclerosis through the suppression of TGF-ß1 expression.

OBJECTIVES: In this study, using an Streptococcus pneumoniae-induced tympanosclerosis (TS) model, we explored the effects of captopril and losartan in the treatment of TS and the possible mechanisms. STUDY DESIGN: A prospective experimental animal study. METHODS: We set up the TS models in both guinea pig and wistar rat by inoculation of type-3 Streptococcus pneumoniae microorganisms and then treated the animals with the combining use of captopril and losartan. Otomicroscopy was employed to observe the development of TS. Auditory brainstem response was used to test the hearing function of animals. Hematoxylin-eosin and von Kossa staining were performed to determine the morphological changes and calcium depositions. The protein expressions of transforming growth factor ß1 (TGF-ß1) were assessed by western blot and immunohistochemistry staining, and the mRNA level of TGF-ß1 was measured by quantitative reverse transcription- polymerase chain reaction. RESULTS: The combining use of captopril and losartan attenuated TS responses in terms of a decrease in the TS incidence and the ABR threshold, a reduction of hyalinization and calcification in the middle ear mucosa and the thickness of the mucosa. In addition, the TGF-ß1 expression was decreased at both protein and mRNA levels. CONCLUSION: Our data indicate, for the first time, that the combining use of captopril and losartan obviously attenuates TS progress through inhibiting the overexpressing of TGF-ß1.