肺表面活性物质对豚鼠分泌性中耳炎的治疗作用

目的:研究肺表面活性物质(PS)对豚鼠分泌性中耳炎(OME)的治疗作用,为OME的治疗探讨新途径。方法:将50只豚鼠随机分为对照组(10只)和实验组(40只),实验组豚鼠采用灭活的肺炎链球菌(KHP)鼓室内注射,建立OME的动物模型(听力下降,中耳积液);模型建立后再将其分为造模组(20耳)、造模对照组(10耳)、治疗组(24耳,向中耳腔注射0.05mlPS)和未治组(23耳,不做任何治疗)。造模5d后,耳内镜下观察鼓膜及鼓室积液的情况,检测各组豚鼠听性脑干反应(ABR)Ⅰ、Ⅲ波潜伏期及反应阈。结果:造模组豚鼠鼓室内注射KHP悬浮液5d后,鼓室出现积液,鼓膜浑浊,光锥消失;ABR反应阈由对照组的(14.00±3.08)dB提高至(45.00±5.67)dB,其差异有统计学意义(P<0.01);治疗组鼓室内注射PS7d后,鼓室积液减少或消失,反应阈由(45.00±5.67)dB降低至(23.53±6.32)dB,分别与造模组及未治组比较,差异有统计学意义(均P<0.01);未治组鼓室积液无变化,鼓膜更加浑浊,反应阈由(45.00±5.67)dB提高至(65.50±6.85)dB。结论:PS对豚鼠OME有治疗作用。     

Effect of systemic clarithromycin and prednisolone on histamine-induced otitis media in guinea pigs

Conclusion: The results of the study showed that clarithromycin has anti-inflammatory and antioxidant effects and, when it is combined with prednisolone, those effects gain strength. Objectives: The present study aims to investigate the effects that the antioxidant and anti-inflammatory activities of clarithromycin and/or prednisolone have on experimental otitis media in effusion-induced guinea-pigs. Method and results: In this study, 35-male guinea pigs were randomly divided into five-groups. For the experimental otitis media, intra-tympanic histamine (0.1 ml) was injected into the guinea pigs in all of the groups except the control group. Then, 24-h after the intra-tympanic injections, clarithromycin (15 mg/kg/day) and/or prednisolone (1 mg/kg/day) were applied intraperitoneally to the guinea-pigs for 7-days. The biochemical analysis showed an increase in antioxidant capacity and a decrease in oxidant status and malondialdehyde (MDA) levels in the clarithromycin group and the prednisolone group and especially in the clarithromycin+prednisolone group, as compared to the experimental group (p < 0.05). In the cytokine analysis, lower levels of interleukin (IL)-6 and IL-17A and higher IL-10 were found in the clarithromycin, prednisolone, and clarithromycin+prednisolone groups than in the experimental group (p < 0.05). Furthermore, the histologic analyses showed histopathologic changes in the middle ear mucosa of the experimental group, but comparatively fewer-histopathologic changes were observed in the clarithromycin, prednisolone, and clarithromycin+prednisolone groups.     

Change of middle ear transfer function in otitis media with effusion model of guinea pigs

Otitis media with effusion (OME) is an inflammatory disease of the middle ear that causes most cases of conductive hearing loss observed in the pediatric population. With the long term goal of evaluating middle ear function with OME, the aim of the current study was to create an animal model of OME in which middle ear transfer functions could be measured. In guinea pigs, OME was created by injecting lipopolysaccharide (LPS) into the middle ear. Evidence of OME was assessed by otoscopy, tympanometry, histology, and by measuring the volume of fluid in the middle ear. Vibrations of the umbo and round window membrane were measured with a laser Doppler vibrometer at frequency range of 200-40kHz in three groups of 3, 7, and 14 days after injection of LPS. Changes in displacement of the umbo and round window membrane in response to 80dB SPL sound in the ear canal were measured across the frequency range. Displacement of both the umbo and round window membrane was reduced at all time points following LPS injections. Further, the change of the displacement transmission ratio (DTR) from the tympanic membrane to the round window occurred mainly in chronic (e.g. 14 days post-LPS injection) OME ears. This study provides useful data for analyzing the change of middle ear transfer function in OME ears.     

可逆性咽鼓管阻塞建立豚鼠急性分泌性中耳炎模型

目的 探索单纯咽鼓管阻塞建立急性分泌性中耳炎豚鼠动物模型的方法,为分泌性中耳炎急性期的相关实验研究提供一种理想的动物模型.方法 健康雄性豚鼠22只,左侧为实验耳,右侧为对照耳,经软腭切开膨胀海绵阻塞左侧咽鼓管咽口建立可逆性分泌性中耳炎动物模型,耳内镜下观察记录鼓膜形态和鼓窜积液情况,分别于术后第7天、第14天及第21天各获取6只造模成功的豚鼠听泡,探查术后膨胀海绵留置情况,HE染色观察病程中中耳黏膜的病理改变.结果 造模组22耳中20耳分别于术后第3～7天出现鼓室浆液性渗出,造模成功率为90.9%,病理改变表现为鼓室黏膜增厚,黏膜下血管扩张,淋巴细胞浸润,近咽鼓管鼓室口部纤毛低矮,数量减少甚至脱落.术后14 d膨胀海绵已基本排出,14～18 d中耳积液消失,同时组织形态逐渐恢复正常.病程中对侧对照耳鼓室无积液,亦无明显病理改变.结论 经软腭切开膨胀海绵咽鼓管阻塞法可成功建立豚鼠急性分泌性中耳炎模型,该分泌性中耳炎可于膨胀海绵自行排出后逐渐自愈.

Abstract：

Objective To develop a animal model for acute otitis media with effusion (OME). Methods In 22 guinea pigs, the left nasal orifice of Eustachian tube was approached via a transpalatal incision and obstructed with polyvinyl acetal material. Right ears were set as the control. Then all the ears were evaluated by otomicroscopy every day. Seven, 14 and 21 days after the intervention, six guinea pigs were killed for histologic study. Results Of the 22 guinea pigs included in this study, 20 ears (90. 9% ) were found to have effusion 3-7 days after the operation, two cases were excluded for purulent otorrhea 10 days postoperatively. The epithelium initially developed hyperplasia, and the submucosa showed vascular and lymphatic dilatations with inflammatory cells infiltration. None of the contralateral control ears had evidence of disease by otomicroscopic examination and histologic study. Conclusion This experimental methods provoked reproducible pathologic characteristics similar to those for otitis media with effusion.     

Pharyngeal ciliary activity in guinea pigs with otitis media

The function of cilia present in the tubotympanum plays an important role in the pathogenesis and chronicity of otitis media with effusion (OME). We used a photoelectric method to examine the ciliary activity in the eustachian tube, in the central pharynx, as well as the nasal cavity of normal animals and animals with otitis media. In normal animals, the ciliary activity in the nasal cavity was not different from that in the eustachian tube. In some animals with otitis media, however, there was a significant difference between the ciliary activity in the nasal cavity and that in the eustachian tube. On the other hand, no significant differences were recognized in any given group of animals examined between the level of the ciliary activity in the central pharynx and that in the eustachian tube. Our findings show that the function of the cilia present in the pharynx is a valid index of that in the eustachian tube. Our study further indicates that it may be possible to clinically assess the ciliary activity in the pharyngeal mucosa biopsied from patients with OME to determine objectively the effects of medical treatments of the disease.     

Pharyngeal ciliary activity in guinea pigs with otitis media

Summary The function of cilia present in the tubotympanum plays an important role in the pathogenesis and chronicity of otitis media with effusion (OME). We used a photoelectric method to examine the ciliary activity in the eustachian tube, in the central pharynx, as well as the nasal cavity of normal animals and animals with otitis media. In normal animals, the ciliary activity in the nasal cavity was not different from that in the eustachian tube. In some animals with otitis media, however, there was a significant difference between the ciliary activity in the nasal cavity and that in the eustachian tube. On the other hand, no significant differences were recognized in any given group of animals examined between the level of the ciliary activity in the central pharynx and that in the eustachian tube. Our findings show that the function of the cilia present in the pharynx is a valid index of that in the eustachian tube. Our study further indicates that it may be possible to clinically assess the ciliary activity in the pharyngeal mucosa biopsied from patients with OME to determine objectively the effects of medical treatments of the disease.     

The effect of otitis media with effusion on perceptual masking

OBJECTIVE: To determine the effect of otitis media with effusion (OME) on perceptual masking (a phenomenon in which spondee threshold for a 2-talker masker is poorer than for a speech-shaped noise masker). DESIGN: Longitudinal testing over a 1-year period following insertion of tympanostomy tubes, using clinical and normal-hearing control groups. SUBJECTS: Forty-seven children having a history of OME were tested. Possible testing intervals were just before the placement of tympanostomy tubes, and up to 3 separate occasions after the placement of the tubes. An age-matched control group of 19 children was tested. METHODS: A perceptual masking paradigm was used to measure the ability of the listener to recognize a spondee in either a speech-shaped noise or a 2-talker masker background. The masker was either continuous or gated on and off with the target spondee. RESULTS: In gated masking conditions, children with a history of normal hearing showed only slight perceptual masking, but the children with a history of OME showed relatively great perceptual masking before surgery and up to 6 months following surgery. In continuous masking conditions, both groups of children showed relatively great perceptual masking and did not differ significantly from each other in this respect either before or after surgery. However, before surgery, the OME group showed higher thresholds in both the 2-talker and speech-shaped noise maskers. CONCLUSIONS: In agreement with previous psychoacoustical findings, the relatively great perceptual masking in gated conditions shown by children with OME history may reflect a general deficit in complex auditory processing.     

Efficacy of transnasal nebulized surfactant on experimental otitis media with effusion in guinea pig

Objective

Recently experimental trials have demonstrated that nebulized natural surfactant is effective in the treatment of experimental otitis media with effusion (OME). Two methodologically incomplete studies using animal models of induced OME have reported that nebulized surfactant can improve Eustachian tube function. There have been no reports that investigated the efficacy of the nebulized surfactant for OME using analysis of tympanic membrane vibration. In this study, we evaluated the therapeutic effect of nebulized natural surfactant by investigation of tympanic membrane vibration.

Materials and methods

The guinea pigs (n = 20) were divided into three groups. Four normal guinea pigs without any treatment were used as normal controls. Sixteen pigs were given experimental OME in both ear and then divided into two groups. The OME model of guinea pig was created by transbullar injection 10 μl of lipopolysaccharide in saline. Experimental group 1 (n = 8) was nebulized phosphate buffered saline (PBS). Experimental group 2 (n = 8) was nebulized natural bovine surfactant. The transnasal nebulization was repeated for 7 days in both experimental groups. On the 8th day, both experimental groups and normal control group were measured the tympanic membrane vibration by laser Doppler vibrometer (LDV), and additionally histopathologic observation was performed by scanning electron microscope (SEM).

Results

The TM vibration velocity in nebulized PBS group showed significantly reduced mainly at low frequencies. However, nebulized natural surfactant group recovered the reduction of tympanic membrane vibration. In nebulized PBS group, SEM showed severe thickened subepithelial layer and hyperplasia of gland structure. However, in nebulized natural surfactant group, SEM showed the reduced the thickness of subepithelial layer and the loss of glandular hyperplasia.

Conclusion

From our results, the nebulized natural surfactant is postulated to be effective in the treatment of intractable OME in animal model. The measurement of tympanic membrane vibration for analysis of middle ear mechanics was significantly corresponding to the middle ear mucosal changes. LDV is a useful tool for investigate the therapeutic effect of nebulized surfactant in experimental OME. With supporting clinical studies, nebulization of natural surfactant may become noninvasive treatment of OME in future.     

Immune-mediated otitis media with effusion

This study was designed to clarify the role of immune reaction in otitis media with effusion (OME). Immune-mediated OME was induced in chinchillas, and characteristics of the disease were evaluated biochemically, cytologically, and histologically. OME was induced by a single intratympanic inoculation of keyhole limpet hemocyanin (KLH) in animals which had a high serum anti-KLH IgG titer. In addition, OME was maintained for 3 weeks in seven of 17 chinchillas, boosted by intradermal and intratympanic injections at 1-week intervals. Cells in the middle ear effusion (MEE) consisted substantially of neutrophils, a few macrophages, and other cells. The mean histamine and prostaglandin E2 levels in the MEE significantly exceeded the levels in corresponding sera. The lining membrane of the bullae was edematous and thickened due to dilatation and rupture of blood capillaries and cell infiltration. Next, the animals were inoculated with immune complex into the tympanic cavity. MEE appeared in all treated ears within three days after injection. Biochemical, cytologic, and histologic findings of the immune complex-induced OME resembled those of immune-mediated OME. An immune peroxidase method demonstrated that neutrophils in MEE phagocytose immune complexes. Our findings suggest that a perpetual cycle of inflammatory substances may maintain an inflammatory condition in the middle ear.     

Naproxen in otitis media with effusion

The levels of prostaglandin E2 (PGE2) were measured in aspirates of the effusion fluid from children suffering from secretory otitis media. The effects of naproxen 5 mg./kg. twice daily for eight weeks on the condition were studied in a placebo-controlled double-blind trial. Nineteen middle-ear aspirates from 13 children, six of whom had bilateral aspirations were examined. Detectable levels of PGE2 (greater than 1.0 ng. per ml.) were present in seven aspirates from seven children and ranged from PGE2 1.0 to 14.4 ng. per ml. In four of the six patients who had bilateral aspirations, PGE2 was detectable in the aspirate from one ear but not the other. However, the severity of otalgia and hearing loss was almost identical in the ears with detectable levels of PGE2 and those without. In a separate trial of naproxen, 11 patients received active drug and 13 received placebo. Naproxen had no significant effect on otalgia or hearing loss compared to placebo. These findings suggest that it is unlikely that non-steroidal anti-inflammatory drugs will be of therapeutic benefit in secretory otitis media.     

Influenza A modification of endotoxin-induced otitis media with effusion in the guinea pig

Summary A total of 120 Hartley strain guinea pigs were used to investigate the possible role of influenza A in endotoxin-induced otitis media with effusion. Intratympanic inoculation of 0.2 ml physiologic saline solution containing 10⁴ plaque-forming units (PFU)/ml of influenza A suspension or 100 ng/ml lipopolysaccharide failed to induce either middle ear effusions or mucociliary pathologies in the tubotymapanum. In contrast, intratympanic inoculation 100 ng/ml endotoxin resulted in prolonged mucociliary dysfunction and middle ear effusions when 0.2 ml 10⁴ PFU/ml of influenza A was inoculated in the tympanic cavity. The inference is drawn that an influenza A infection might predispose the middle ear to endotoxin-induced otitis media with effusion.This paper was presented in part at the Fifth International Symposium on Recent Advances in Otitis Media in Ft. Lauderdale, Florida, USA, on 20–24 May 1991     

Nitrotyrosine in otitis media with effusion

OBJECTIVES: Our purpose was to investigate mucosal cell injury due to the nitric oxide (NO)-superoxide system in otitis media with effusion. METHODS: We determined the levels of nitrotyrosine (NT) and NO and the activities of superoxide dismutase (SOD) and lactic dehydrogenase (LDH) in 90 middle ear fluid samples. RESULTS: The NT concentration was significantly higher in group A (<16 years old) than in group C (>50 years old; p < .05), and significantly higher in the acute group than in the chronic group (p < .05). The NO concentration did not show a significant difference among the groups. The activity of SOD showed significant correlations with the concentrations of NT and NO and with LDH activity (p < .05). The LDH activity was significantly greater in group A than in group C (p < .05). CONCLUSIONS: Our results indicate involvement of the NO-superoxide system in the pathogenesis of otitis media with effusion, showing evidence of protein and/or cell injury in the middle ear.     

Hearing acuity in otitis media with effusion and adhesive otitis media

The relationship between findings established by the photograph of the tympanic membrane and hypacusia was determined by pure-tone audiometry. Subjects were 352 ears of 267 patients diagnosed as otitis media with effusion (OME) or adhesive otitis media (AdOM). These two diseases were evaluated on the basis of photographs of the tympanic membrane taken from 1983 to 1988. The results were as follows; 1) The effect of aging process on hearing acuity was noted in patients with these diseases, i.e., values of hearing acuity by air and bone conduction were, in descending order, early-, mature-, and advanced-age groups, with significant differences (p less than 0.01). 2) Greater loss of acuity by air conduction in the middle to high frequency ranges was found in patients with OME than with AdOM. In bone conduction, a decrease in low frequency range in the early--and mature--age groups was larger in patients with AdOM than with OME and a significant decrease in 2000-6000 Hz in the advanced-age group was more marked in patients with OME than with AdOM (p less than 0.05). 3) Among patients with AdOM, those with retained fluid in the middle ear showed a significant decrease in air conduction values and a significant increase in air-bone gap compared with those without fluid (p less than 0.05). 4) The degrees of adhesion correlated well with a decrease in hearing acuity. 5) Hearing acuity was not influenced by attic retraction, white plaque, scar and atrophy of their tympanic membranes.     

Histopathology of otitis media with effusion

Summary Ultrastructural studies of the middle ear mucosa appear to be of significant value in better understanding the pathology of otitis media with effusion (OME). Our present study was undertaken in order to take advantage of the use of electron microscopy in investigating all areas of the middle ear mucosa. Tissues studied were obtained from the fresh postmortem temporal bones of three patients with OME and terminal head and neck malignancies. In the mucoid type of effusion (cases 1 and 2), goblet cells were seen to proliferate and secretory activity was greatly enhanced. In contrast, there was no evidence of secretory cell proliferation in the serous type of effusion. It was noteworthy that accumulated fluid was not homogeneous in the same ear, as exemplified by case 1, in which both mucoid and serous effusions were present. This occurrence was possibly the result of topographic diversity involving the secretory activity of the middle ear.Presented at the Extraordinary International Symposium on Recent Advances in Otitis Media with Effusion, Kyoto, Japan, 12–15 January 1985     

Bacteriology of otitis media with effusion

A study was undertaken to evaluate the prevalence and antibiotic susceptibility of bacteria present in the middle ear of patients with otitis media with effusion. Middle ear effusions (MEE), nasopharyngeal and throat swabs were obtained at operation and cultured for aerobic and anaerobic bacteria. Two hundred and fifty-nine effusions were obtained from 152 subjects examined. Haemophilus influenzae was isolated from 32 (12.3 per cent) effusions, Streptococcus pneumoniae from seven (2.7 per cent), Staphylococcus aureus from seven (2.7 per cent), Branhamella catarrhalis from one (0.4 per cent)--Group A beta haemolytic streptococci from one (0.4 per cent) and Staphylococcus epidermidis from three (1.9 per cent). The occurrence of respiratory pathogens in MEE reflected their prevalence in the upper respiratory tract. Significantly fewer children who had received antibiotics prior to surgery had organisms present in the MEE. Eight and a half per cent of H. influenzae and 64 per cent of B. catarrhalis were resistant to ampicillin. The present study confirms that bacteria are present in the middle ear in a significant number of patients with otitis media with effusion.     

Balance and otitis media with effusion

Dizziness can be caused by a variety of peripheral vestibular, central, and systemic disease processes. Eustachian tube dysfunction with and without middle-ear effusion has been considered one of the most common causes of balance disturbances in young children. Several studies have indicated that during an episode of otitis media the child's balance deteriorates and the child may become clumsy and fall more often. Thus, not only the adverse effect on hearing should be considered in the management of a child with otitis media, but also the child's balance.     

豚鼠慢性化脓性中耳炎鼓室硬化模型

目的 研究建立慢性化脓性中耳炎鼓室硬化动物模型豚鼠的方法,为慢性化脓性中耳炎鼓室硬化的动物实验研究提供一种有用的动物模型.方法 采用小鼠腹腔连续传代3次增强毒力的金黄色葡萄球菌,采取反复中耳腔注射的方法,制备豚鼠慢性化脓性中耳炎动物模型,了解化脓性中耳炎对听阈的影响及是否导致鼓室硬化.结果 将钙化斑及局部骨化作为鼓室硬化的标志,接种细菌后1周就可观察到中耳粘连的出现,接种后2周出现钙化斑等鼓室硬化灶,听力较对照耳明显下降.观察到硬化灶最常见的部分是下鼓室,其次是鼓室上隐窝、上鼓室、鼓膜、听骨链间隙和咽鼓管鼓口.结论 豚鼠慢性化脓性中耳炎鼓室硬化动物模型病理变化与人慢性化脓性中耳炎鼓室硬化类似,是研究慢性化脓性中耳炎鼓室硬化较好的动物模型.     

Ultrastructural changes of the cochlea in guinea pigs with acute bacterial otitis media

Acute bacterial otitis media was induced in guinea pigs with H. influenzae to investigate the ultrastructural changes of the organ of Corti. The results showed that the stereocilia in the organ of Corti adjacent to the round window was disoriented and partially disappeared, there was an increase in the number of lysosomes and marked aggregation of smooth endoplasmic reticula, the surface of the supporting cells was damaged and broken. The most prominent damages were observed on specimens 7 days following the middle ear inoculation with H. influenzae. The damages to the supporting cells were repaired 21 days after the inoculation though the damaged stereocilia did not show any sign of recovery.