Argyrophilic nucleolar organizer regions in auditory meatal skin and middle ear cholesteatoma

Comparative silver-staining of argyrophilic nuclear organizer regions (AgNORs) was performed to study the proliferative activity of auditory meatal skin and middle ear cholesteatoma. AgNOR expression patterns were counted by standardized methods in two centres, Bochum and London, and mean numbers of dots per nucleus were calculated. Specimens of normal auditory meatal skin showed 1.54 dots/nucleus (n = 12) in the Bochum study, whereas cholesteatoma had 3.71 dots/nucleus (n = 21). In the London study normal meatal skin showed two dots/nucleus (n = 4), whereas acquired cholesteatoma (n = 8) gave a mean of 4.90 dots/nucleus and congenital cholesteatoma a mean of 4.70 dots/nucleus (n = 2). Our findings confirm the hyperproliferative state of middle ear cholesteatoma, suggest that the congenital variety of cholesteatoma may have a similar activity and indicate that AgNOR staining is a useful technique for assessing cellular proliferation in cholesteatoma and objectifying and quantifying its aggressive behaviour.     

Apoptosis in meatal skin,cholesteatoma and squamous cell carcinoma of the ear

Apoptosis in normal meatal skin, cholesteatoma and squamous cell carcinoma (SCC) of the ear was investigated by using morphological analysis and in-situ specific labelling of fragmented DNA. In meatal skin, apoptotic cells were localized mainly in the granular layers, but were not so restricted in cholesteatoma, while in SCC they were even more dispersed. The apoptotic index (AI) was low (1.59 ± 0.10 sem ) in normal skin. It was increased in cholesteatoma (2.09 ± 0.11), and was intermediate in SCC (1.72 ± 0.14). By contrast, the mitotic index (MI) increased from 0.19 ± 0.02 in normal skin, to 0.25 ± 0.01 in cholesteatoma and to 0.25 ± 0.02 in SCC. Our findings indicate that apoptosis is involved in the epithelial homeostasis of meatal skin, cholesteatoma and SCC of the ear. The hyperproliferation of epithelial cells in cholesteatoma is counteracted by an increased apoptosis rate, while in SCC the increased cell proliferation without a compensatory increase in apoptosis may be associated with the malignant transformation.     

Canal wall-down procedure with soft posterior meatal wall reconstruction in acquired cholesteatoma: focus on postoperative middle ear status*

Objectives: We reviewed surgical results of canal wall-down tympanoplasty (CWDT) with soft posterior meatal wall reconstruction (SWR) for acquired cholesteatoma (AC), and identified factors associated with surgical outcomes.

Methods: Results from 119 ears with AC (pars flaccida, n?=?99; pars tensa, n?=?20) that underwent CWDT with SWR were retrospectively reviewed. We defined postoperative balloon-like retraction (PBR) with web formation, which needed reoperation to clean accumulated cerumen, as postoperative deep retraction pocket (PDRP).

Results: Residual cholesteatoma was found in 11 ears (9.2%). Seven residual cholesteatomas were treated with outpatient operation. Seven ears (5.9%) showed PDRP. A transcanal approach was applied to all PDRPs. Postoperative mastoid reaeration was observed in 57 ears (47.9%). No factors significantly associated with residual cholesteatoma or PDRP were identified. The frequency of postoperative mastoid reaeration was significantly higher among cases with young age (<50 years), stage I cholesteatoma, or type I ossiculoplasty.

Conclusion: CWDT with SWR showed low rates of residual cholesteatoma or postoperative deep retraction pocket (PDRP). Most residual cholesteatomas and PDRPs could be dealt with using a minimally invasive procedure. Young age, stage I cholesteatoma, and type I ossiculoplasty were associated with postoperative mastoid reaeration. This procedure seems fully feasible for surgical treatment of AC.     

Colony-stimulating factor in middle ear cholesteatoma

Granulocyte-macrophage colony stimulating factor (GM-CSF) was found in human middle ear cholesteatoma tissues by immunohistochemical technique using mouse monoclonal anti-(human) GM-CSF IgG. Immunofluorescent staining showed the presence of GM-CSF in the peribasal area, in some suprabasal cells of the epithelium, and in the inflammatory connective tissue, especially in the monocytes and fibroblasts. These findings were confirmed by the immunoperoxidase method. Staining of the external ear canal epithelium, however, was significantly weaker than that of the cholesteatoma epithelium. The presence of GM-CSF in cholesteatoma appears to be a response to inflammation occurring in the middle ear cavity. Moreover, our in vitro study showed that GM-CSF induced the proliferation and protein synthesis of basal keratinocytes. This study suggests that GM-CSF is involved in the development and destructive effects of middle ear cholesteatoma.     

p53 expression and keratinocyte hyperproliferation in middle ear cholesteatoma1

Keratinocytes in middle ear cholesteatoma have hyperproliferative properties. There is controversy regarding the role of p53 and its effect on cellular proliferation in cholesteatoma. This study was instituted to examine this. Cholesteatoma and deep meatal skin control specimens were analysed for MIB‐1 (n = 7, controls = 7), a marker of cellular proliferation, and p53 (n = 17, controls = 17) expression by immunocytochemistry. Expression of p53 was minimal or absent in both cholesteatoma and controls (P = 0.2). MIB‐1 expression was higher, but not significantly so, in cholesteatoma than in controls (P = 0.09). Our study has shown no significant p53 expression in cholesteatoma epithelium. This suggests that there is no dysfunction in the p53‐mediated cell cycle control mechanisms in cholesteatoma.     

10例先天性中耳胆脂瘤临床分析

目的：探讨先天性中耳胆脂瘤的临床特征及手术方法。方法：回顾性分析10例先天性中耳胆脂瘤患者的临床资料。5例经耳内途径行鼓室探查并一期鼓室成形术,3例行闭合式乳突根治术及鼓室成形术,2例行开放式乳突根治术及鼓室成形术。结果：术中见5例胆脂瘤局限于中后鼓室,3例位于中鼓室及上鼓室,2例病变范围广泛,侵及乳突。术后6个月平均纯音听阈为30dBHL,气骨导差在20dB以内,复查颞骨CT均未发现胆脂瘤残留和复发。结论：先天性中耳胆脂瘤病变隐匿,常于鼓室前方或后方,易破坏听骨链,导致传导性聋;影像学检查可为诊断及术式选择提供依据;早期手术治疗可获得较好的听力重建效果。     

Matrix metalloproteinase-2 and Matrix metalloproteinase-9 in cholesteatoma and deep meatal skin

A qualitative and quantitative study of the presence of Matrix metalloproteinase-2 (MMP-2) and Matrix metalloproteinase-9 (MMP-9) in cholesteatoma was performed. Ten cholesteatoma and four deep meatal skin specimens were analysed for gelatinase activity at molecular weights corresponding to MMP-2 and MMP-9 using Sodium Dodecyl Sulphate Polyacrylamide Gel Electrophoresis (SDS PAGE) Zymography. Gelatinase activity at 72 kDa and 92 kDa was investigated. Western blotting was employed using primary monoclonal antibodies to provide a qualitative assessment of MMP-2 and MMP-9. Non-parametric data analysis using the Mann–Whitney U test did not show a significant difference in expression of MMP-2 (P = 0.51) or MMP-9 (P = 0.14) between the two tissue types. Western blotting showed the presence of both MMP-2 and MMP-9 in the majority of specimens, both cholesteatoma and deep meatal skin.     

肿瘤坏死因子受体1在中耳胆脂瘤上皮中的表达

目的 研究肿瘤坏死因子受体1（TNFR1）在中耳继发性胆脂瘤上皮的表达，探讨其在胆脂瘤型中耳炎的发病机制中的作用。方法 分别采用免疫组化法和Western blot技术检测TNFR1在36例中耳胆脂瘤组织及20例正常外耳道皮肤组织中的表达情况。结果 TNFR1在36例胆脂瘤上皮各层细胞中均有强表达，定位于胞膜和胞质，而外耳道皮肤表达较弱甚至没有表达，TNFR1在胆脂瘤上皮中的蛋白表达水平较外耳道皮肤显著增高，差异有统计学意义（P〈0．01）。结论 胆脂瘤上皮中TNFR1表达较外耳道皮肤显著增高，肿瘤坏死因子可能通过与TNFR1相结合而导致胆脂瘤上皮细胞的过度增殖及凋亡。     

中耳胆脂瘤的遗传学研究进展

中耳胆脂瘤是一类位于鼓室和/或乳突内的团块,由角化的鳞状上皮细胞、上皮下的结缔组织以及角化碎片构成,伴/不伴炎症反应。临床表现主要为耳流脓和听力下降,当侵袭周围组织结构时则可产生眩晕、周围性面瘫、颅内感染等一系列颅内外并发症,甚至危及生命。手术是目前唯一有效的治疗方式。其病因及发病机制仍未完全清楚。随着全外显子测序等新一代测序技术的发展,中耳胆脂瘤的遗传学研究取得了一些突破性进展。近年来,关于胆脂瘤的家族聚集性、基因突变及其与综合征的关系的相关研究越来越多,本文将对上述内容进行综述,从而为中耳胆脂瘤的非手术治疗、药物研发提供参考。     

An immunocytochemical study of Matrixmetalloproteinase 1 in cholesteatoma and deep meatal skin

Aim: To investigate the hypothesis that bacterial toxins produced in the middle ear during acute otitis media (AOM), travel via the round window membrane (RWM) to cause cochlear damage and hence sensorineural hearing loss (SNHL). Methods: We investigated toxins produced by the two most common pathogens isolated from middle ear effusions in children. These were Pneumolysin, a product of S. pneumoniae and a cytoplasmic extract of Haemophilus influenza. Both electrophysiological measurements and morphological investigations were made after these toxins were placed on the RWM of guinea pigs. Control experiments were done using artificial perilymph as a substitute for the toxins. In each experiment one of the cochleas served as an internal control. Results: Losses in both the Cochlear Microphonics (CM) and Compound Action Potential (CAP) were observed indicating impaired hair cell function resulting in SNHL. Morphological studies showed hair cell destruction. Examination of the RWM exposed to pneumolysin showed disruption of this three-layered structure. Conclusions: The RWM is a probable portal for access to the inner ear by substances produced during AOM. Sensorineural hearing loss could result from the type of damage observed. More experiments are needed to verify this hypothesis and to determine the extent of damage that can be caused by these substances, especially with regard to the position of the damaged hair cells. Most of the damage observed occurred in the basal turn. Careful audiological examination (especially at high frequencies) of children who suffer with AOM might reveal a population of children with sensorineural hearing loss which might otherwise go undetected.     

上皮细胞增殖和凋亡在中耳胆脂瘤发病学的意义(附33例报告)

目的 :通过探讨胆脂瘤上皮 (CE)、胆脂瘤患者外耳道上皮 (CAMS)和正常健康者外耳道上皮(NAMS)的增殖和凋亡及 p5 3基因与上皮细胞增生和凋亡的关系 ,阐明增殖和凋亡异常在胆脂瘤发病中的作用。方法 :采用免疫组织化学和原位末端标记技术检测细胞增殖标记物 PCNA、Ki- 6 7和 p5 3蛋白在 33例 CE、2 5例CAMS和 10例 NAMS细胞的表达及细胞凋亡。结果 :在上述标本中均有 PCNA表达及凋亡细胞存在 ,但不同类型上皮阳性细胞数量、染色强度及分布不同。 CE存在过度增殖和凋亡 ,CAMS亦存在过度增殖。 p5 3表达与PCNA呈正相关 ,与细胞凋亡呈负相关。结论 :CE具有过度增殖和凋亡的特性 ,增殖与凋亡的紊乱与胆脂瘤形成密切相关。     

Presence of osteoclasts in middle ear cholesteatoma: a study of undecalcified bone sections

Conclusions: Osteoclasts are unlikely to be involved in bone resorption in middle ear cholesteatoma.

Objective: The authors searched for osteoclasts in undecalcified bone sections in patients with middle ear cholesteatoma to determine whether and to what extent these cells are involved in this disease.

Methods: Twelve patients, eight men and four women, aged 30–87 years, who underwent tympanomastoidectomy were enrolled. Six patients had primary acquired middle ear cholesteatoma (cholesteatoma group) and the other six patients had other otologic diseases including otosclerosis, non-cholesteatomatous chronic otitis media, adhesive otitis media, perilymphatic fistula and ossicular malformation (control group). The scutum bone was collected during surgery, fixed with ethanol, stained with Villanueva bone stain, and embedded in methyl methacrylate. Five-micrometer-thick sections were prepared and examined under a polarizing microscope. Images were analyzed using a semiautomatic graphics system.

Results: No osteoclasts were seen in any of the samples in either group. To avoid the risk of under-estimating the presence of osteoclasts, the number of osteoclasts was considered to be <1 in each sample, and the osteoclast density was calculated. The osteoclast densities in both the cholesteatoma and control groups were significantly lower than the sex- and age-matched standard value of the normal iliac cortical bone (p?=?.028).     

TNF-R2 expression in acquired middle ear cholesteatoma

Acquired middle ear cholesteatoma is a disease which promotes bone erosion resulting in potentially serious complications. The tumor necrosis factor alpha (TNF-α) is present in cholesteatoma and it is related to bone erosion, as shown by different authors. To understand the aggressiveness characteristics of cholesteatoma is necessary, however, to better address the presence and distribution of their receptors.ObjectiveTo evaluate the expression of type 2 TNF-α receptor (TNF-R2) in fragments of cholesteatoma and correlate it to the degree of inflammation present.Material and methodsobservational cross-sectional study, which analyzed 33 fragments of cholesteatomas through histological analysis and immunohistochemistry (using as primary antibody to TNF-R2 LabVision ® brand). The evaluation was performed by means of a qualitative and semi-quantitative agreement with the observed intensity. For statistical analysis we used the Fisher exact test and Spearman's correlation coefficient (considered statistically significant when p ≤ 0.05).ResultsThe expression of TNF-R2 was present in all fragments, however a statistical analysis showed no correlation or association between inflammation and the expression of TNF-R2.ConclusionsTNF-R2 is present in cholesteatoma of the middle ear, however, its expression is not directly related to the degree of inflammation observed in patients with this disease.     

Management of acquired cholesteatoma of the middle ear and the mastoid by combined approach tympanoplasty: a long-term view

Mastoid cavities following surgery for cholesteatoma of the middle ear and the mastoid can be subject to recurrent infections, life-long attendance for cavity cleaning and restrictions in social activity. These problems may be avoided with a successful combined approach tympanoplasty. One hundred and forty-one patients (151 ears) treated by combined approach tympanoplasty, followed over a period ranging from 5 to 23 yr were analysed. The results are presented with an average follow-up of 14.5 yr. A fixed retraction pocket, recurrent cholesteatoma or conversion to a cavity was regarded as a failure. Failures and evidence of future failures in the form of fixed retraction pockets occurred in the first 5 yr. By not including cases with a follow-up period of less than 5 yr, we have attempted to achieve stable long-term results, 73.5% of the cases were successful. The success rate could be improved further and should provide an acceptable method of treatment.     

继发性胆脂瘤中CCL27的定位及检测

目的 明确趋化因子CCL27在继发性中耳胆脂瘤中的病理学表达,探讨可能的临床意义。 方法 应用免疫组织化学SP法检测CCL27在继发性中耳胆脂瘤(n=30)及正常耳后皮肤组织(n=30)的表达情况,并分析该趋化因子与胆脂瘤病理学之间的联系。 结果 继发性胆脂瘤、耳后皮肤组织中CCL27的阳性率分别为6.7%和33.3%,差异有统计学意义(P=0.021)。 结论 继发性胆脂瘤组织中CCL27的低表达可能与胆脂瘤的增殖过于活跃有关,CCL27可能在继发性中耳胆脂瘤发病过程中扮演重要角色。     

儿童中耳胆脂瘤诊治进展

儿童中耳胆脂瘤在患病率、发病机制与临床表现等方面都具有人群的特殊性.其起病隐匿,早期诊治成为临床难点.随着对儿童中耳胆脂瘤认识的提高和普及,临床中对此疾病的关注度明显提高.临床分期的应用及手术治疗的精准性趋于完善,伴有腭裂、特纳综合征、唐氏综合症等特殊人群的中耳胆脂瘤患儿的远期预后管理也得到了重视.     

小龄儿童全耳内镜中耳胆脂瘤手术可行性分析

目的 探讨小龄儿童经耳道全耳内镜中耳胆脂瘤手术可行性,提示临床诊疗路径.方法 收集2016年4月-2021年3月经耳道全耳内镜手术治疗的小龄(≤6岁)中耳胆脂瘤患儿为研究对象.入组26例患儿,男19例,女7例;年龄1.9～6岁,平均年龄(4.1±0.9)岁.对病例一般资料、病变程度、术中情况、术后并发症、住院时间、复发...