The effect of zoledronic acid on middle ear osteoporosis: An animal study.

Hearing function in older patients may be related to bone structure. We conducted an experiment to evaluate the effect of zoledronic acid on osteoporotic middle ear ossicles in an animal model. Our subjects were 19 female New Zealand white rabbits (38 ears) weighing 2 to 4 kg. We divided the rabbits into three groups: one group consisted of 6 rabbits with osteoporotic ears that were treated with zoledronic acid; the second group was made up of 8 rabbits with osteoporotic ears that were not treated; a control group included 5 rabbits with normal ears that were untreated. After an oophorectomy, the 6 study rabbits were administered 0.1 ml/kg of zoledronic acid intravenously. All rabbits were sacrificed 16 weeks later, and the middle ear ossicles were removed for investigation under light microscopy. Although osteoporosis enhanced the osteoclastic bone resorption of the ossicles, zoledronic acid enhanced osteoblastic activity on osteoporotic middle ear ossicles. The incidence of osteoporosis was 93.8% in the untreated osteoporosis group and 33.3% in zoledronic acid group-a statistically significant difference (OR: 0.24; 95% CI: 0.09 to 0.58; p < 0.001). Osteoporosis appears to increase the resorption of the middle ear ossicles, a process that can be avoided with zoledronic acid administration. Prevention of the effects of osteoporosis in humans may help decrease the irreversible changes in the middle ear ossicles.

Iatrogenic Phenol Injury Causing Facial Paralysis With Tympanic Membrane and Ossicular Necrosis.

OBJECTIVE: To describe a serious iatrogenic injury and propose means of reducing the risk of its reoccurrence. PATIENTS: A 21-year-old man who suffered facial paralysis, complete necrosis of the tympanic membrane, and ossicular discontinuity because of chemical burn from accidental application of copious amounts of topical anesthetic phenol into the ear. INTERVENTIONS: Conservative management of facial paralysis and delayed reconstruction of the tympanic membrane and ossicular chain. MAIN OUTCOME MEASURES: Gradual recovery to grade 1/6 facial function, successful repair of the tympanic membrane, but persistent 30-dB conductive hearing loss after partial ossicular replacement prosthesis presumably because of scarring. CONCLUSION: Phenol is a highly toxic chemical, topically to both skin and eyes. Absorbed through the skin it can have lethal cardiotoxicity. It is also potent neurotoxin at concentrations much lower (4-7%) than used for tympanic membrane anesthesia (89%) and has long been used therapeutically to destroy nerves in patients of contractions or intractable pain. Otologists need to have a healthy respect for the dangers of using phenol. As only a minute quantity is needed for tympanic anesthesia, commercially available prepackaged applicators are preferred. Storage of stock bottles of 89% phenol solutions in clinical settings risks injury to both patients and practitioners.

Bone conduction activation through soft tissues following complete immobilization of the ossicular chain, stapes footplate and round window.

Classically it has been thought that bone conduction activation at the mastoid leads to relative motion between the stapes footplate and the oval window due to inertial and to compression (distortion) mechanisms. However, several recent clinical findings and experimental manipulations may point to additional mechanisms. These manipulations were extended in the present study. In ten fat sand rats, following obliteration of one ear, auditory nerve brainstem evoked response (ABR) thresholds were recorded in response to broad band click stimuli, either air conducted (AC) through insert earphones or bone conducted (BC) delivered directly to the exposed skull bone. Following this, the entire ossicular chain, stapes footplate and round window were completely immobilized with super glue, leading to a mean AC threshold elevation of 44 dB, but to a mean BC threshold change (elevation) of only 3.5 dB. In this state of complete immobilization, the bone vibrator was applied to a pool of saline in the surgical area and ABR was elicited with a mean threshold which was not significantly different from that of the BC threshold. When the bone vibrator was then applied to the eye without touching the bone at the orbit, the resulting ABR threshold was about 20 dB greater than the BC threshold. In conclusion, BC stimulation can activate the cochlea without two mobile windows. Furthermore, the cochlea can be activated by a fluid pathway and by application of a bone vibrator to non-osseous sites (soft tissue conduction).

Roles of an anti-tuberculosis medication and surgery in patients with tuberculous otitis media.

CONCLUSION: The standard treatment for tuberculous otitis media (TOM) without complications consists of anti-tuberculosis (anti-TB) medication, with which we experienced good treatment outcomes. However, surgery is required for recovery of anatomy and hearing function. OBJECTIVE: To determine the clinical characteristics of TOM that might optimize diagnosis and to evaluate the differences in clinical courses between patients treated with and without surgery. METHODS: We analyzed 14 patients (16 ears) who had been diagnosed and treated for TOM. Radiologic findings, laboratory data, and audiometry results were also evaluated. Patients were divided into a chemotherapy group and a surgery group according to treatment modality. RESULTS: Temporal bone CT (TBCT) showed total occupation of the tympanic cavity by soft tissue and little evidence of ossicular erosion. In the chemotherapy group, dry ears were obtained in all but one patient (14 ears) after treatment. Normalized tympanic membrane (TM) was found in 50% in the chemotherapy group and in 75% in the surgery group. The air-bone gap (ABG) changed from 40.3 +/- 2.5 dB to 47.0 +/- 19.2 dB in the chemotherapy group and from 35.2 +/- 7.6 dB to 30.2 +/- 11.4 dB in the surgery group. After treatment, ABG improved by > 10 dB in one ear in the chemotherapy group and in four ears in the surgery group.

Effects of cadmium on the hearing system.

The functional resemblance between kidney proximal tubular and inner ear epithelial cells which has often been pointed out in the literature led us to hypothesize that nephrotoxic agents that cause renal tubular injury might also impair the function of inner ear cells. As one of the most toxic environmental nephrotoxic agents is cadmium, we aimed to study its effects on hearing experimentally in rats. In this study, increased blood and renal cortical cadmium levels were associated with high cadmium accumulation in ear ossicles and labyrinth in rats exposed to cadmium. The changes in auditory brainstem response (ABR) and otoacoustic emission in 2-month-old male rats exposed to drinking water containing 5 and 15 ppm CdCl2 for 30 days showed that cadmium-induced nephrotoxicity was associated with signs of defective hearing at a concentration of 15 ppm CdCl2 but that 5 ppm CdCl2 caused hearing loss without affecting kidney function. The mean latency of ABR wave 1, which indicates the function of the cochlea, was 1.335 +/- 0.31 ms in the control group and 1.641 +/- 0.052 and 1.74 +/- 0.88 ms in the rats subjected to 5 and 15 ppm CdCl2, respectively (p < 0.001). In the cadmium-treated groups short interpeak wave I-III latencies (p < 0.01) indicated cochlear dysfunction and this was also supported by the distortion product otoacoustic emission results (p < 0.001). Non-significant changes in wave III and V latencies were accepted as evidence of unaltered function of the other parts of the auditory system. These results suggest that hair cells are more sensitive to cadmium than kidney tubule cells and that the cochlear component of hearing is more vulnerable to cadmium toxicity than other parts of the auditory system.

Biomechanical properties of sterilized human auditory ossicles.

Bone allograft material is treated with sterilization methods to prevent the transmission of diseases from the donor to the recipient. The effect of some of these treatments on the integrity of the bone is unknown. This study was performed to evaluate the effect of several sterilization methods on the mechanical behaviour of human middle ear bones. Due to the size and composition of the bones (approximately 1.5 mm diameter by 4 mm long), mechanical testing options were limited to the traditional platens compression test. Experiments were first performed with synthetic bone to evaluate the precision of this test applied to small specimens. Following this, fresh frozen human ossicles were thawed and sterilized with (i) 1 N NaOH (n = 12); (ii) 0.9% LpH, a phenolic solution (n = 12); or (iii) steam at 134 degrees C (n = 18). A group of 26 control specimens did not receive any sterilization treatment. Material and structural properties were determined from axial compression testing. Results from the synthetic bone showed that the test was reproducible, with standard deviations less than 20% of the means. Significant differences occurred in stiffness and ultimate force values between NaOH-treated and autoclaved bones when compared to normals (p<0.05), but not for LpH-treated bones. LpH is not approved for medical use, so NaOH is the most appropriate of the treatments studied for the sterilization of ossicle allografts.

Correlation between loss of middle ear bones and altered goosecoid gene expression in the branchial region following retinoic acid treatment of mouse embryos in vivo.

The homeobox gene goosecoid marks the Spemann organizer in vertebrate gastrula embryos, and is expressed in the craniofacial region, body wall and limbs during organogenesis. Mouse mutants of goosecoid displayed a variety of phenotypes related to the expression pattern at mid-embryogenesis. These defects included loss of the tympanic ring and malformation of the malleus, phenotypes which were reminiscent of the teratogenic effects of retinoic acid (RA). Here we investigated the correlation of goosecoid gene expression and RA-teratogenicity following treatment of mouse embryos in vivo at embryonic day (E) 8 + 5 h. We found that goosecoid was specifically affected at E10.5 in branchial arches I and II. Expression was either reduced to background levels or restricted to the branchial cleft region. This change in goosecoid gene expression correlated with a loss of middle ear ossicles and a partial or complete deletion of the tympanic ring, suggesting a role for goosecoid in executing the RA teratogenic effects.

Lack of chondrotoxicity of ofloxacin otic solution on the auditory ossicle cartilages of juvenile guinea pigs.

The effect of 0.3% otic solution of ofloxacin on the cartilages constituting the epiphyses of the auditory ossicles and the wall of the auditory tube was histologically examined after 30-day repeated intratympanic administration to juvenile male guinea pigs aged 4 weeks. Ofloxacin showed no chondrotoxicity for the cartilages and, in some but not all animals, reduced haemorrhage and neutrophil infiltration in the tympanic cavity, compared with that of the control. Further, the articular cartilage of the humeral trochlea and femoral condyle also showed no change.

Teratological studies on craniofacial malformations.

Craniofacial malformations cause great human suffering. The purpose of the experimental studies was to investigate teratogenically induced craniofacial malformations in the rat, and to study if vitamin B6 could prevent the teratogenically induced malformations in the rat. The aim of the clinical investigation was to compare mandibulofacial dysostosis (MFD) with hemifacial microsomia (HFM) and thalidomide-induced malformations restricted to the first and second branchial arches. In the experimental studies we used two different teratogenic agents, etretinate and BAPN (beta-aminoproprionitrile). Vitamin B6 was administered one day prior to and simultaneously with the teratogenic agent. The induced malformations were observed by direct microscopy, histology, differential staining, microdissection and enzyme histochemistry. Knowledge of isoenzymic differentiation was obtained by isoelectric focusing and polyacrylamide gel electrophoresis. The clinical features of 29 patients with MFD, 26 with HFM and seven with thalidomide-induced malformations were investigated. The patients underwent clinical investigations, radiography, tomography, computed tomography, surgical exploration and audiograms. The etretinate-induced syndrome in the rat shows similarities to first and second branchial arch syndromes in man. Defective formation of Meckel's cartilage and the cartilaginous skull base, the zygoma and the middle ear ossicles were prominent features of the observed malformations. The induced malformations were accompanied by increased staining for alkaline phosphatase (APase) in the skull and skull base cartilages and Meckel's cartilage. BAPN induced cleft palate in 95% of the cases and the teratogenically induced cleft palate was accompanied by a pathological differentiation pattern that could be traced by determination of isoenzymes in the palatal shelves as well as in amniotic fluid. Vitamin B6 could prevent the teratogenic malformations induced by etretinate and BAPN in the rat. Comparing MFD, HFM and thalidomide-induced malformations, all syndromes included patients with external, middle and inner ear malformations. Cranial nerve palsy/paresis was only seen in HFM and thalidomide-induced malformations. A relationship between disturbed neural crest cell migration and defects of the first and second branchial arches seems possible.

Middle ear anomalies induced by hypertriazene administration in the mouse.

The middle ear is derived from various embryonic tissues. Many experiments using teratogens have been performed, employing the difference of each tissue's sensitivity to the teratogen and the tissue's critical time of development. Triazene, a foliate metabolism antagonist, produced anomalies in fetuses that resembled those associated with thalidomide in humans, the so-called the first and second branchial syndrome. In our experiment, we administrated 3,3-dimethyl-1-phenyltriazene, an inductor of triazene, to pregnant mice at 7 to 14 days of gestation resulting in unique fetal anomalies. We examined the development of the stapes, stapedial artery, facial nerve, and oval window using an optical microscopic and three-dimensional reconstruction. Middle ear and facial nerve anomalies in mice depend on the gestation day when triazene is administrated. The stapedial artery, oval window, facial nerve (horizontal segment), stapes footplate, and styloid process are affected on the 9th to 11th administration day, the annular stapedialis on the 10th to 11th day, and the malleus and incus on the 9th to 11th day. The use of the Vox View/Mac, allowed us to create three-dimensional pictures from two-dimensional slides providing an improved understanding of the relationships between anatomical structures.

Effect of endotoxin on cultured rat middle ear epithelium, rat meatal epidermis, and human keratinocytes.

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.

Ossicular erosion by cholesteatoma: investigation by scanning electron microscopy utilizing a new preparation technique.

A new technique for removing the soft tissues from middle ear ossicles to prepare them for scanning electron microscopy is described. The technique involved use of a solution of hypochlorite. The bony surface of normal ossicles was studied as a control group after preparation by the technique and no morphological distortion was observed. Ossicles eroded by cholesteatoma were then studied, and we propose that the erosion occurs in three stages; pumicing, pitting and cavitating. The mucoperiosteum of normal ossicles and otosclerotic foci were also studied.

Effect of flavonoid on PGE2-induced alterations in percentage collagen synthesis in ossicle organ cultures.

In adult guinea-pig stapes organ cultures, 3H (2,3)-proline incorporation into the collagenase-digestible protein (CDP) and non-collagen protein (NCP) fractions of the ossicles was measured and the percentage of collagen synthesis (PCS) was calculated. Prostaglandin E2 (PGE2), a potent stimulator of bone resorption, inhibited the PCS in low concentrations (5 and 25 microM), whereas it stimulated it in pharmacological concentrations (50 and 100 microM). Ipriflavon, an isoflavone derivative of therapeutical potential in otosclerosis, also reduced the PCS in 1, 10 and 50 microM concentrations. 50 microM Ipriflavon stimulated the PCS inhibited by 5 microM PGE2, but decreased the PGE2-induced PCS enhancement in vitro.

The effects of an adhesive in the middle ear.

This study investigated the effects of enbucrilate (Histoacryl) on the middle ear. Small amounts of the adhesive were used in 81 middle-ear operations performed on 50 rabbits. Light microscopic investigation of these ears 1, 2, and 3 months after surgery showed bone necrosis and a granulation tissue reaction surrounded by foreign-body giant cells. Inner-ear changes were observed in a third of the cases. Therefore, enbucrilate should only be used with extreme care and in minute quantities and never near the labyrinthine capsule.

[Experimental studies on the question of the enzymatic destruction of the ossicular bones during middle ear inflammations (author's transl)]. / Experimentelle Untersuchungen zur Frage der enzymbedingten Gehörknöchelchendestruktion im Verlauf einer chronischen Mittelohrentzündung

The proteolytic activity of purulent middle ear secretions from patients with cholesteatoma, chronic otitis media, acute otitis media and radical mastoid cavities were investigated. The proteolytic activity in the secretions from cases with cholesteatoma is higher than from patients with other chronic otitis media. The collagenolytic effect of the matrix of cholesteatoma combined with the high level of the bacterial and leukocytic proteinases in the middle ear secretions is undoubtedly one of the main factors in the destruction of the ossicular bones.