Sustained-Release Triamcinolone Acetonide Hydrogels Reduce Hearing Threshold Shifts in a Model for Cochlear Implantation with Hearing Preservation.

INTRODUCTION: In recent years, the preservation of residual hearing has become a major factor in patients undergoing cochlear implantation (CI). In studies attempting to pharmaceutically improve hearing preservation rates, glucocorticoids (GCs) applied perioperatively in many institutions have emerged as a promising treatment regimen. Although dexamethasone is most commonly used and has been applied successfully by various research groups, recently pharmacological properties have been reported to be relatively unsuitable for topical delivery to the inner ear. Consequently other glucocorticoids merit further evaluation. The aim of this study was therefore to evaluate the otoprotective effects of the topical application of a sustained-release triamcinolone acetonide (TAAC) hydrogel in CI with hearing preservation. METHODS: Normal-hearing pigmented guinea pigs were randomized into a group receiving a single dose of a 6% TAAC poloxamer 407 hydrogel, a group receiving a 30% TAAC hydrogel and a control group. All hydrogel applications were performed 1 day prior to CI. After a cochleostomy was drilled, a specifically designed silicone electrode was inserted into the scala tympani for 5 mm. Frequency-specific compound action potentials of the auditory nerve (0.5-32 kHz) were measured pre- and directly postoperatively as well as on days 3, 7, 14, 21, and 28. Finally, temporal bones were harvested for histological evaluation. RESULTS: Application of the TAAC hydrogels resulted in significantly reduced hearing threshold shifts in low, middle and high frequencies and improved spiral ganglion cell survival in the second turn of the cochlea. Outer hair cell numbers in the basal and second turn of the cochlea were slightly reduced after TAAC application. CONCLUSION: In summary, we were able to demonstrate functional benefits of a single preoperative application of a TAAC hydrogel in a guinea pig model for CI, which persisted until the end of the observational period, that is, 28 days after surgery.

Evaluation of Sustained-Release Steroid Hydrogels in a Guinea Pig Model for Noise-Induced Hearing Loss.

The otoprotective effects of thermoreversible poloxamer 407 hydrogels containing dexamethasone or triamcinolone acetonide were evaluated in an animal model of noise-induced hearing loss. Seven days after noise exposure, hearing threshold shifts at 16 kHz were significantly reduced in the 6% dexamethasone group (p < 0.05). Even though no significant differences in hair cell counts were found, histological analysis revealed a significantly higher spiral ganglion cell density in the first turn of the cochlea in this group (p < 0.05). No otoprotective effects were observed after the application of the triamcinolone acetonide hydrogels. As the findings of this study indicate potential otoprotective effects of sustained topical dexamethasone delivery in the setting of noise-induced hearing loss, this strategy merits further evaluation.

Noise trauma and systemic application of the selective glucocorticoid receptor modulator compound A.

BACKGROUND: Selective glucocorticoid receptor modulators (SEGRMs) comprise a novel class of drugs promising both reduced side effects and similar pharmacological potency relative to glucocorticoids, which presently serve as the only clinical treatment for many otologic disorders. In the first otologic SEGRM experiment in an animal model of noise trauma, we compare the effects of Compound A (a SEGRM) and dexamethasone (potent glucocorticoid). METHODS: Forty adult guinea pigs received experimental treatment once daily for ten days. The animals were divided into four cohorts based on the treatment received: Compound A (1 mg/kg or 3 mg/kg), dexamethasone (1 mg/kg) as gold standard, or water as negative control. After five applications, animals were exposed to broadband noise (8-16 kHz) at 115 dB for three hours. Hearing thresholds were determined by recording auditory brainstem responses to clicks and noise bursts (1-32 kHz) and were assessed a week prior to and immediately after exposure, as well as on days 1, 3, 7, 14, 21, and 28. Cochleae were prepared as whole-mounts or embedded and sectioned for histological analysis. RESULTS: Relative to the control treatments, Compound A failed to preserve auditory thresholds post-noise exposure with statistical significance. Histological analyses confirm the physiological result. CONCLUSION: The present findings suggest that Compound A does not have substantial otoprotective capacities in a noise trauma model.

Sustained release of triamcinolone acetonide from an intratympanically applied hydrogel designed for the delivery of high glucocorticoid doses.

The pharmacokinetic properties and tolerability of a triamcinolone acetonide poloxamer 407 hydrogel for intratympanic application were investigated in a guinea pig model. Evaluation of in vivo release kinetics showed very high initial perilymph drug levels, with clinically relevant levels present for a minimum of 10 days. Assessment of auditory brainstem response thresholds showed a minimal, delayed and transient threshold shift, which was apparent on day 3 and resolved by day 10. No relevant histological changes of the middle and inner ear structures were noted, and hair cell counts showed no significant differences between treated and untreated ears. Thus, the triamcinolone-acetonide-loaded poloxamer 407 hydrogel is an effective vehicle for sustained high-dose inner ear glucocorticoid delivery.

Arterial Blood Supply of the Stifle Joint in Horses.

The vascularization pattern of the equine stifle joint is insufficiently described in the literature, even though there is a growing need for knowledge of the exact blood supply, as (i) arthroscopy and endoscopic surgery techniques are increasingly performed in horses and (ii) ex vivo models of menisci need nutrient supply that mimic the in vivo situation. The aim of this study was to describe the vessels involved in the stifle joint supply and the exact branching pattern of the popliteal artery. Colored latex was injected into the arteries of nine pelvic limbs of equine cadavers (n = 6) to evaluate the occurrences, variations and approximate diameters of vessels that supplied the stifle joints. Next to a branch of the saphenous and descending genicular arteries, eleven branches of the popliteal artery could be described in horses that feed the vascular network of the stifle joint. With a focus on the blood supply of the menisci, a vascularization map was created to show the main influx to these intra-articular structures in detail. These findings are potentially of great importance to both clinicians in preparation of best-suited incisions for arthroscopy and researchers designing new approaches for meniscal studies and choosing suitable animal models.

Effects of prenatal stress on hypothalamic-pituitary-adrenal (HPA) axis function over two generations of guinea pigs (Cavia aperea f. porcellus).

Prenatal stress can alter hypothalamic-pituitary-adrenal axis function with potential consequences for later life. The aim of our study was to examine in guinea pigs (Cavia aperea f. porcellus) the effects of stress experienced during F0 pregnancy on glucocorticoid levels in plasma and feces, as well as challenge performance, in F1 offspring (n=44) and fecal glucocorticoid levels in F2 offspring (n=67). F1 animals were either born to F0 dams that had been stressed with strobe light during early to mid pregnancy, resulting in a short term increase but long-term down-regulation of maternal glucocorticoid levels, or to undisturbed F0 dams. The same stressor was used as a challenge for F1 offspring at age 26 days and around 100 days. Basal plasma cortisol concentrations during early F1 development, as well as overall glucocorticoid levels at challenge tests, were lower in F1 animals that were prenatally stressed than in control animals. Fecal cortisol metabolites were initially at lower levels in prenatally stressed F1 animals, relative to control animals, but shifted to higher levels around day 68, with an additional sex difference. Effects were also seen in the F2 generation, as male but not female offspring of prenatally stressed F1 animals had significantly higher levels of cortisol metabolites in feces after weaning. We conclude that stress exposure of F0 dams resulted in lower basal glucocorticoid levels in F1 offspring during the pre-pubertal phase and during stress exposure, but higher glucocorticoid levels in post-adolescent F1 animals. Also in males of F2 generation effects of stress exposure of F0 dams were detected.

Granulosa cell tumour in a harbour porpoise (Phocoena phocoena) from German waters.

Neoplastic diseases in harbour porpoises Phocoena phocoena have rarely been described, and there are no reported gonadal stromal tumours. A 12 yr old female harbour porpoise was stranded on the North Sea coast of Schleswig-Holstein, Germany. Necropsy findings included a severe granulomatous pneumonia, pregnancy and a left ovarian tumour. Respiratory insufficiency was the likely cause of death. There was a multinodular mass composed of cords with peripherally palisading cells within the left ovary. The histological and cytological appearance of the neoplasm was suggestive of a granulosa cell tumour; supportive immunohistochemical stains, including those for vimentin, cytokeration, carcinoembryonic antigen, c-kit, chromogranin and α-smooth muscle action, were negative.

Muscle Fibre Architecture of Thoracic and Lumbar Longissimus Dorsi Muscle in the Horse.

As the longissimus dorsi muscle is the largest muscle in the equine back, it has great influence on the stability of the spine and facilitates proper locomotion. The longissimus muscle provides support to the saddle and rider and thereby influences performance in the horse. Muscular dysfunction has been associated with back disorders and decline of performance. In general, muscle function is determined by its specific intramuscular architecture. However, only limited three-dimensional metrical data are available for the inner organisation of the equine longissimus dorsi muscle. Therefore, we aimed at investigating the inner architecure of the equine longissimus. The thoracic and lumbar longissimus muscles of five formalin-fixed cadaveric horse backs of different ages and body types were dissected layerwise from cranial to caudal. Three-dimensional coordinates along individual muscle fibre bundles were recorded using a digitisation tool (MicroScribe®), to capture their origin, insertion and general orientation. Together with skeletal data from computed tomography (CT) scans, 3D models were created using imaging software (Amira). For further analysis, the muscle was divided into functional compartments during preparation and morphometric parameters, such as the muscle fascicle length, pennation angles to the sagittal and horizontal planes, muscle volume and the physiological cross-sectional area (PCSA), were determined. Fascicle length showed the highest values in the thoracic region and decreased from cranial to caudal, with the cranial lumbar compartment showing about 75% of cranial fascicle length, while in most caudal compartments, fascicle length was less than 50% of the fascicle length in thoracic compartments. The pennation angles to the horizontal plane show that there are differences between compartments. In most cranial compartments, fascicles almost run parallel to the horizontal plane (mean angle 0°), while in the caudal compartment, the angles increase up to a mean angle of 38°. Pennation angles to the sagittal plane varied not only between compartments but also within compartments. While in the thoracic compartments, the fascicles run nearly parallel to the spine, in the caudal compartments, the mean angles range from 0-22°. The muscle volume ranged from 1350 cm3 to 4700 cm3 depending on body size. The PCSA ranged from 219 cm2 to 700 cm2 depending on the muscle volume and mean fascicle length. In addition to predictable individual differences in size parameters, there are obvious systemic differences within the muscle architecture along the longissimus muscle which may affect its contraction behaviour. The obtained muscle data lay the anatomical basis for a specific biomechanical model of the longissimus muscle, to simulate muscle function under varying conditions and in comparison to other species.

Long-term effects and potential limits of intratympanic dexamethasone-loaded hydrogels combined with dexamethasone-eluting cochlear electrodes in a low-insertion trauma Guinea pig model.

Cochlear implantation has become the most effective hearing restoration method and is one of the great advances in modern medicine. Early implants have been continuously developed into more efficient devices, and electro-acoustic stimulation is increasingly expanding the indication criteria for cochlear implants to patients with more residual hearing. Therefore, protecting the cochlear structures and maintaining its intrinsic capacities like residual hearing has become more important than ever before. In the present study, we aimed to assess the long-term protective effects of a dexamethasone-eluting electrode combined with the preoperative intratympanic application of a dexamethasone-loaded thermoreversible hydrogel in a cochlear implant guinea pig model. 40 normal-hearing animals were equally randomized into a control group receiving an unloaded hydrogel and a non-eluting electrode, a group receiving a dexamethasone-loaded hydrogel and a non-eluting electrode, a group receiving an unloaded hydrogel and a dexamethasone-eluting electrode and a group receiving both a dexamethasone-loaded hydrogel and a dexamethasone-eluting electrode. Residual hearing and impedances were investigated during a period of 120 days. Tissue response and histological changes of cochlear structures were analyzed at the end of the experiments. Treatment with dexamethasone did not show a significant protective effect on residual hearing independent of treatment group. Although the majority of the cochleae didn't exhibit any signs of electrode insertion trauma, a small degree of tissue response could be observed in all animals without a significant difference between the groups. Foreign body giant cells and osteogenesis were significantly associated with tissue response. Hair cells, synapsin-1-positive cells and spiral ganglion cells were preserved in all study groups. Cochlear implantation using a dexamethasone-eluting electrode alone and in combination with a dexamethasone-loaded hydrogel significantly protected auditory nerve fibers on day 120. Post-implantation impedances were equal across study groups and remained stable over the duration of the experiment. In this study we were able to show that use of a dexamethasone-eluting electrode alone and in combination with preoperative application of dexamethasone-loaded hydrogel significantly protects auditory nerve fibers. Furthermore, we have shown that a cochlear implantation-associated hearing threshold shift and tissue response may not be completely prevented by the sole application of dexamethasone.

Effects of sustained release dexamethasone hydrogels in hearing preservation cochlear implantation.

It has been shown that glucocorticoids reduce the hearing threshold shifts associated with cochlear implantation. Previous studies evaluated the administration of glucocorticoids immediately before surgery or the repeated pre- or perioperative systemic application of glucocorticoids. The aim of this study was to evaluate the effects of a sustained release dexamethasone hydrogel in hearing preservation cochlear implantation. To address this issue, a guinea pig model of cochlear implantation was used. 30 normal hearing pigmented guinea pigs were randomized into a group receiving a single dose of a dexamethasone/poloxamer407 hydrogel one day prior to surgery, a second group receiving the hydrogel seven days prior to surgery and a control group. A silicone cochlear implant electrode designed for the use in guinea pigs was inserted to a depth of 5 mm through a cochleostomy. Compound action potentials of the auditory nerve (frequency range 0.5-32 kHz) were measured preoperatively, directly postoperatively and on postoperative days 3, 7, 14, 21 and 28. Following the last audiometry, temporal bones were harvested and histologically evaluated. Dexamethasone hydrogel application one day prior to surgery resulted in significantly reduced hearing threshold shifts at low, middle and high frequencies measured at postoperative day 28 (p < 0.05). Application of the hydrogel seven days prior to surgery did not show such an effect. Dexamethasone application one day prior to surgery resulted in increased outer hair cell counts in the cochlear apex and in reduced spiral ganglion cell counts in the basal and middle turn of the cochlea, a finding that was associated with a higher rate of electrode translocation in this group. In this study, we were able to demonstrate functional benefits of a single preoperative intratympanic application of a sustained release dexamethasone hydrogel in a guinea pig model of cochlear implantation.

Evaluation of the selective glucocorticoid receptor agonist compound A for ototoxic effects.

OBJECTIVES/HYPOTHESIS: To evaluate the selective glucocorticoid receptor agonist (SEGRA) compound A, a potential novel therapeutic for inner ear disorders, for ototoxic effects. STUDY DESIGN: Laboratory animal study. METHODS: Experimental guinea pigs were grouped as follows: Systemic application of compound A (1.5 mg/kg and 4.5 mg/kg; n = 6/group) and intratympanic application of compound A (1 mM and 10 mM; n = 6/group). Contralateral ears in topically treated animals served as controls. Hearing thresholds were determined by auditory brainstem response before and directly after the application of compound A, as well as on days 3, 7, 14, 21, and 28. At the end of the experiments, temporal bones were harvested for histological evaluation. RESULTS: Systemic administration of compound A (1.5 mg/kg and 4.5 mg/kg) did not cause hearing threshold shifts, whereas the intratympanic injection (1 mM and 10 mM) resulted in a hearing loss. Histological analysis of the middle and inner ears after topical compound A application showed alterations in the tympanic membranes, the auditory ossicles, and the round window membranes, whereas spiral ganglion cells and hair cells were not affected. CONCLUSION: SEGRAs such as compound A could provide novel therapeutic options for the treatment of inner ear disorders and reduce metabolic side effects. Whereas the intratympanic application of compound A resulted in a hearing loss, the systemic application of compound A merits evaluation for otoprotective effects in trauma models.

Effects of chronic stress during pregnancy on maternal performance in the guinea pig (Cavia aperea f. porcellus).

Stress experienced during pregnancy can have persistent impact on the female's physiology and behaviour not only during but even beyond pregnancy. The present study aimed to evaluate such long-term effects of stress in terms of repeated strobe light exposure during early to mid gestation on behavioural aspects of mothering activities and lactational effort in lactating guinea pigs. We found that maternal behaviour was negatively affected by stress experience during pregnancy with treatment females developing a higher level of offspring-directed aggression than controls. In addition, our measure of lactational performances showed tendencies of lowered milk supply and longer pup suckling durations in stressed females. We suggest that this may represent a strategy to advance infant weaning following demanding conditions caused by chronic stress experience during pregnancy.

Sex-specific impact of prenatal stress on growth and reproductive parameters of guinea pigs.

Body condition and reproductive maturation are parameters of reproductive success that are influenced by sexual hormones rising in the circulation during the time of puberty. Various endocrine systems can be programmed by conditions experienced during early life. Stress for instance is supposed to be capable of influencing fetal development, leading to adjustments of offspring's later physiology. We examined whether prenatal stress (induced by exposure to strobe light) during early- to mid-gestation was capable of affecting later reproductive parameters in guinea pigs (Cavia aperea f. porcellus). Therefore, we measured the levels of testosterone and progesterone from the age of day 12-124 in prenatally stressed (PS, n = 20) and unaffected control animals (n = 24). Furthermore, we determined the timing of puberty and growth. Body weight development revealed significantly faster growth in PS females compared to control animals. The onset of first estrus was slightly earlier in PS females, however not significantly so. Cycle lengths and levels of progesterone differed between groups over the course of time with higher progesterone levels and more constant cycles among PS females compared to control females who displayed marked differences between first and subsequent cycles. Levels of testosterone did not differ between groups. We conclude that prenatal stress accelerates growth and maturity in females, but not in males.

Hair cortisol: a parameter of chronic stress? Insights from a radiometabolism study in guinea pigs.

Measurement of hair cortisol has become popular in the evaluation of chronic stress in various species. However, a sound validation is still missing. Therefore, deposition of radioactivity in hair and excretion into feces and urine after repeated injection of (3)H-cortisol was studied in guinea pigs (n = 8). Each animal was given intraperitoneally 243.6 kBq (3)H-cortisol/day on 3 successive days. After the first injection, all voided excreta were collected for 3 days. After the second injection, hair was shaved off the animals' back and newly grown hair was obtained on day 7. Following methanol extraction, radiolabeled and unlabeled glucocorticoid metabolites (GCM) in fecal and hair samples were characterized by high-performance liquid chromatography (HPLC) and enzyme immunoassays (EIA). In feces, maximum radioactivity was reached 8 h (median) post each injection, whereas maxima in urine were detected in the first samples (median 2.5 h). Metabolites excreted into feces (13.3% ± 3.7) or urine (86.7%) returned nearly to background levels. HPLC of fecal extracts showed minor variation between individuals and sexes. In hair, small amounts of radioactivity were present. However, two EIAs detected large amounts of unlabeled GCM, including high levels at the position of the cortisol standard; radioactivity was absent in this fraction, demonstrating that (3)H-cortisol was metabolized. Furthermore, large amounts of immunoreactivity coinciding with a radioactive peak at the elution position of cortisone were found. These results show for the first time that only small amounts of systemically administered radioactive glucocorticoids are deposited in hair of guinea pigs, while measurement of large amounts of unlabeled GCM strongly suggests local production of glucocorticoids in hair follicles.

Chronic stress in pregnant guinea pigs (Cavia aperea f. porcellus) attenuates long-term stress hormone levels and body weight gain, but not reproductive output.

Stress, when extreme or chronic, can have a negative impact on health and survival of mammals. This is especially true for females during reproduction when self-maintenance and investment in offspring simultaneously challenge energy turnover. Therefore, we investigated the effects of repeated stress during early- and mid-gestation on the maternal stress axis, body weight gain and reproductive output. Female guinea pigs (Cavia aperea f. porcellus, n = 14) were either stressed (treatment: exposure to strobe light in an unfamiliar environment on gestational day -7, 0, 7, 14, 21, 28, 35, 42) or left completely undisturbed (control) throughout pregnancy. Females of both groups received the same respective diets, and reproductive parameters were evaluated upon parturition. Additionally, hormonal data were obtained from blood and feces. The stress exposure induced a significant increase in plasma cortisol concentrations during the afternoon. In contrast to this short-term response in plasma cortisol concentrations, we found no significant differences in the levels of cortisol metabolites in feces collected after stress exposure between groups and even significantly decreased levels of fecal cortisol metabolites on non-stress days over time in treatment females. Among treatment females, gain in body weight was attenuated over gestation and body weight was lower compared to control females during lactation, especially in cases of large litter sizes. No differences could be seen in the reproductive parameters. We conclude that repeated stress exposure with strobe light during early- and mid-gestation results in a down-regulation of the hypothalamic-pituitary-adrenal axis and lower weight gain in treatment females, but has no effect on reproductive output.