Biomedical subjects

Robert K Shepherd

Publications and source records attributed to Robert K Shepherd.

16 recordsLinked to original sources

PubMed2006

Effect of interphase gap and pulse duration on electrically evoked potentials is correlated with auditory nerve survival.

We investigated the effect of pulse duration (PD) and interphase-gap (IPG) on the electrically-evoked auditory brain stem response (EABR) and viiith nerve compound action potential (ECAP) of deafened guinea pigs in order to test the hypothesis that the extent of change in these neural responses is affected by the histological status of the auditory nerve. Fifteen guinea pigs were deafened by co-administration of kanamycin and furosemide. Animals were acutely implanted with an 8-band electrode array at 1, 4 or 12 weeks following deafening. EABR and ECAP input/output functions were recorded in response to charge balanced biphasic current pulses. We determined the change in current required to equalize; (i) the EABR amplitude when the duration of the current pulse was doubled (104-208 micros/phase); and (ii) the EABR and ECAP amplitudes when the IPG was increased from 8 to 58 micros using a 104 micros/phase current pulse. Following the completion of each experiment the cochleae were examined quantitatively for spiral ganglion neuron survival. As expected, the current level required to evoke an EABR with equal amplitude was lower when the animal was stimulated with current pulses of 208 compared with 104 micros/phase. Moreover, the current level required to evoke EABR/ECAPs with equal amplitude was lower when current pulses had an IPG of 58 versus 8 micros. Importantly, there was a reduction in the magnitude of this effect with greater neural loss; the reduced efficacy of changing both PD and IPG on these electrically-evoked potentials was statistically correlated with neural survival. These results may provide a tool for investigating the contribution of auditory nerve survival to clinical performance among cochlear implant subjects.

Action Potentials↗

PubMed2006

Pneumococcal meningitis threshold model: A potential tool to assess infectious risk of new or existing inner ear surgical interventions.

HYPOTHESIS: A minimal threshold of Streptococcus pneumoniae is required to induce meningitis in healthy animals for intraperitoneal (hematogenous), middle ear, and inner ear inoculations, and this threshold may be altered via recent inner ear surgery. BACKGROUND: There has been an increase in the number of reported cases of cochlear implant-related pneumococcal meningitis since 2002. The pathogenesis of pneumococcal meningitis is complex and not completely understood. The bacteria can reach the central nervous system (CNS) from the upper respiratory tract mucosa via either hematogenous route or via the inner ear. The establishment of a threshold model for all potential routes of infection to the CNS in animals without cochlear implantation is an important first step to help us understand the pathogenesis of the disease in animals with cochlear implantation. METHODS: Fifty-four otologically normal adult Hooded Wistar rats (27 receiving cochleostomy and 27 controls) were inoculated with different amounts of bacterial counts via three different routes (intraperitoneal, middle ear, and inner ear). Rats were monitored during 5 days for signs of meningitis. Blood, cerebrospinal fluid, and middle ear swabs were taken for bacterial culture, and brains and cochleae were examined for signs of infection. RESULTS: The threshold of bacterial counts required to induce meningitis is lowest in rats receiving direct inner ear inoculation compared with both intraperitoneal and middle ear inoculation. There is no change in threshold between the group of rats with cochleostomy and the control (Fisher's exact test, p < 0.05). CONCLUSION: A minimal threshold of bacteria is required to induce meningitis in healthy animals and is different for three different routes of infection (intraperitoneal, middle ear, and inner ear). Cochleostomy performed 4 weeks before the inoculation did not reduce the threshold of bacteria required for meningitis in all three infectious routes. This threshold model will also serve as a valuable tool, assisting clinicians to quantitatively analyze if the presence of a cochlear implant or other CNS prostheses alter the risk of meningitis.

Animals↗

PubMed2006

Pneumococcal meningitis: development of a new animal model.

HYPOTHESIS: The rat is a suitable animal to establish a model for the study of pneumococcal meningitis postcochlear implantation. BACKGROUND: There has been an increase in the number of cases of cochlear implant-related meningitis. The most common organism identified was Streptococcus pneumoniae. Whether cochlear implantation increases the risk of pneumococcal meningitis in healthy subjects without other risk factors remains to be determined. Previous animal studies do not focus on the pathogenesis and risk of pneumococcal meningitis postimplantation and are based on relatively small animal numbers, making it difficult to assess the cause-and-effect relationship. There is, therefore, a need to develop a new animal model allowing direct examination of the pathogenesis of meningitis in the presence of a cochlear implant. METHODS: Eighteen nonimplanted rats were infected with 1 x 10 and 1 x 10 colony-forming units (CFU) of a clinical isolate of S. pneumoniae via three different inoculation routes (middle ear, inner ear, and i.p.) to examine for evidence of meningitis during 24 hours. Six implanted rats were infected with the highest amount of bacteria possible for each route of inoculation (4 x 10 CFU i.p., 3 x 10 CFU middle ear, and 1 x 10 CFU inner ear) to examine for evidence of meningitis with the presence of an implant. The histological pattern of cochlear infections for each of the three different inoculating routes were examined. RESULTS: Pneumococcal meningitis was evident in all 6 implanted animals for each of the three different routes of inoculation. Once in the inner ear, bacteria were found to enter the central nervous system via either the cochlear aqueduct or canaliculi perforantes of the osseous spiral lamina, reaching the perineural and perivascular space then the internal acoustic meatus. The rate, extent, and pattern of infection within the cochleae depended on the route of inoculation. Finally, there was no evidence of pneumococcal meningitis observed in 18 nonimplanted rats inoculated at a lower concentration of S. pneumoniae when observed for 24 hours postinoculation. CONCLUSION: Meningitis in implanted rats after inoculation with a clinical isolate of S. pneumoniae is possible via all three potential routes of infection via the upper respiratory tract. The lack of meningitis observed in the 18 nonimplanted rats suggests that longer postinoculation monitoring periods are required to ensure whether or not meningitis will develop. Based on this work, we have developed a new animal model that will allow quantitative risk assessment of meningitis postcochlear implantation, and the assessment of the efficacy of potential interventional strategies in future studies.

Animals↗

PubMed2006

Protective effects of local administration of ciprofloxacin on the risk of pneumococcal meningitis after cochlear implantation.

OBJECTIVES: To determine whether ciprofloxacin retains its antimicrobial activity after storage with Healon at ambient temperature and at 37 degrees C over 5 weeks and then to establish whether the application of ciprofloxacin/Healon onto scala tympani electrode arrays reduces the risk of meningitis in implanted rats inoculated with S. pneumoniae. STUDY DESIGN: In vitro laboratory and in vivo animal studies METHODS: The antibacterial activity of three concentrations of ciprofloxacin/Healon (7.5, 75, and 750 microg/mL) was examined over 5 weeks at both ambient temperature (23 degrees C) and body temperature (37 degrees C). Thirty-six rats (18 implanted with ciprofloxacin [750 mg/mL]/Healon-coated electrode array and 18 without the coating) were infected with S. pneumoniae 4 weeks after implantation by way of three different routes of infection (hematogenous, middle ear, and inner ear) and observed for the development of meningitis. RESULTS: The antibacterial activity of ciprofloxacin/Healon was maintained over 5 weeks at both 23 degrees C and 37 degrees C. The implanted rats with the ciprofloxacin/Healon-coated electrode array were protected from meningitis when the bacteria were given by way of the hematogenous route (Fisher's exact test, P = .008) but not when the bacteria were inoculated directly into the middle or inner ear. However, the time to develop meningitis was significantly longer in rats implanted with a coated array, irrespective of the route of inoculation (P < .05, log rank test). CONCLUSION: Our animal model demonstrated that a ciprofloxacin-coated electrode array can protect healthy implanted rats from meningitis when the route of infection is hematogenous and can delay the onset of meningitis when bacteria are inoculated directly into the middle or inner ear.

Adjuvants, Immunologic↗

PubMed2006

Basis of electrical stimulation of the cochlea and the cochlear nucleus.

Sensorineural hearing loss is the most common form of deafness in humans. In patients with a severe-profound sensorineural hearing loss therapeutic intervention can only be achieved by direct electrical stimulation of the auditory nerve via a cochlear implant, or - in cases where a cochlear implant is not a surgical option - neurons within the central auditory pathway via an auditory brainstem implant. This paper reviews the basis of electrical stimulation of these structures with an emphasis on pathophysiology and safety.

Animals↗

PubMed2006

Aminoglycoside-induced degeneration of adult spiral ganglion neurons involves differential modulation of tyrosine kinase B and p75 neurotrophin receptor signaling.

Aminoglycoside antibiotics induce sensorineural hearing loss by destroying hair cells of the organ of Corti, causing progressive secondary degeneration of primary auditory or spiral ganglion neurons (SGNs). Recent studies show that the p75 neurotrophin receptor (NTR) is aberrantly up-regulated under pathological conditions when the neurotrophin receptor tyrosine kinases (Trks) are presumptively down-regulated. We provide in vivo evidence demonstrating that degenerating SGNs induced an augmented p75NTR expression and a coincident reduction of TrkB expression in their peripheral processes. Nuclear transcription factors c-Jun and cyclic AMP response element-binding protein phosphorylated by p75NTR- and TrkB-activated signal pathways, respectively, also showed a corresponding differential modulation, suggesting an activation of apoptotic pathways, coupled to a loss of pro-survival neurotrophic support. Our findings identified brain-derived neurotrophic factor (BDNF) expression in hair and supporting cells of the adult cochlea, and its loss, specifically the mature form, would impair TrkB-induced signaling. The precursor of BDNF (pro-BDNF) is differentially cleaved in aminoglycoside-deafened cochleae, resulting in a predominant up-regulation of a truncated form of pro-BDNF, which colocalized with p75NTR-expressing SGN fibers. Together, these data suggest that an antagonistic interplay of p75NTR and TrkB receptor signaling, possibly modulated by selective BDNF processing, mediates SGN death in vivo.

Aminoglycosides↗

PubMed2005

Chronic depolarization enhances the trophic effects of brain-derived neurotrophic factor in rescuing auditory neurons following a sensorineural hearing loss.

The development and maintenance of spiral ganglion neurons (SGNs) appears to be supported by both neural activity and neurotrophins. Removal of this support leads to their gradual degeneration. Here, we examined whether the exogenous delivery of the neurotrophin brain-derived neurotrophic factor (BDNF) in concert with electrical stimulation (ES) provides a greater protective effect than delivery of BDNF alone in vivo. The left cochlea of profoundly deafened guinea pigs was implanted with an electrode array and drug-delivery system. BDNF or artificial perilymph (AP) was delivered continuously for 28 days. ES induced neural activity in two cohorts (BDNF/ES and AP/ES), and control animals received BDNF or AP without ES (BDNF/- and AP/-). The right cochleae of the animals served as deafened untreated controls. Electrically evoked auditory brainstem responses (EABRs) were recorded immediately following surgery and at completion of the drug-delivery period. AP/ES and AP/- cohorts showed an increase in EABR threshold over the implantation period, whereas both BDNF cohorts exhibited a reduction in threshold (P < 0.001, t-test). Changes in neural sensitivity were complemented by significant differences in both SGN survival and soma area. BDNF cohorts demonstrated a significant trophic or survival advantage and larger soma area compared with AP-treated and deafened control cochleae; this advantage was greatest in the base of the cochlea. ES significantly enhanced the survival effects of BDNF throughout the majority of the cochlea (P < 0.05, Bonferroni's t-test), although there was no evidence of trophic support provided by ES alone. Cotreatment of SGNs with BDNF and ES provides a substantial functional and trophic advantage; this treatment may have important implications for neural prostheses.

Animals↗

PubMed2005

Survival of partially differentiated mouse embryonic stem cells in the scala media of the guinea pig cochlea.

The low regenerative capacity of the hair cells of the mammalian inner ear is a major obstacle for functional recovery following sensorineural hearing loss. A potential treatment is to replace damaged tissue by transplantation of stem cells. To test this approach, undifferentiated and partially differentiated mouse embryonic stem (ES) cells were delivered into the scala media of the deafened guinea pig cochlea. Transplanted cells survived in the scala media for a postoperative period of at least nine weeks, evidenced by histochemical and direct fluorescent detection of enhanced green fluorescent protein (EGFP). Transplanted cells were discovered near the spiral ligament and stria vascularis in the endolymph fluid of the scala media. In some cases, cells were observed close to the damaged organ of Corti structure. There was no evidence of significant immunological rejection of the implanted ES cells despite the absence of immunosuppression. Our surgical approach allowed efficient delivery of ES cells to the scala media while preserving the delicate structures of the cochlea. This is the first report of the survival of partially differentiated ES cells in the scala media of the mammalian cochlea, and it provides support for the potential of cell-based therapies for sensorineural hearing impairment.

Animals↗

PubMed2005

Cochlear implantation in rats: a new surgical approach.

The laboratory rat has been used extensively in auditory research but has had limited use in cochlear implant related research due mainly to the surgically restricted access to the scala tympani. We have developed a new surgical method for cochlear implantation in rats. The key to this protocol was cauterizing the stapedial artery (SA) and making a small cochleostomy near the round window in order to enlarge the surgical access to the scala tympani. Five normal hearing Hooded Wistar rats were used to investigate the effect of cauterizing the SA on hearing and auditory nerve survival. Results showed that cauterizing the SA was surgically feasible, afforded excellent exposure of the round window niche for cochleostomy, and did not adversely affect acoustic thresholds measured electrophysiologically. Moreover, there was no difference in spiral ganglion cell densities for any cochlear turn when compared with the contralateral control ears. Three deafened rats were subsequently implanted with a scala tympani electrode array using this new surgical approach. Electrically evoked auditory brainstem responses using bipolar stimulation, and subsequent cochlear histopathology demonstrated that cochlear implantation using a custom-made rat electrode array was safe and effective. The surgical approach presented in this paper presents a safe and effective procedure for acute or chronic cochlear implantation in the rat model.

Animals↗

PubMed2005

Exogenous BDNF rescues rat spiral ganglion neurons in vivo.

OBJECTIVE: To determine if exogenous neurotrophins can prevent spiral ganglion neuron degeneration in the rat cochlea. BACKGROUND: The loss of hair cells resulting in sensorineural hearing loss also leads to the secondary degeneration of spiral ganglion neurons. The effectiveness of cochlear implantation in patients with profound sensorineural hearing loss relies in part on the survival of spiral ganglion neurons; therefore, any therapy that can prevent or halt the loss of these neurons would be of potential clinical benefit. Previous research has shown that intracochlear infusion with neurotrophins can provide trophic support to SGNs in deafened guinea pigs. Whether this effect is seen in other species remains to be determined. METHODS: After documenting the rate of spiral ganglion neuron degeneration after ototoxic deafening, we investigated the trophic effects of exogenous brain-derived neurotrophic factor (BDNF) on rat spiral ganglion neurons. The left cochleae of profoundly deafened rats were implanted with a drug delivery system connected to a mini-osmotic pump. BDNF or artificial perilymph was infused for 28 days; then the cochleae were prepared for histological study. RESULTS: Treatment with BDNF led to a statistically significant increase in spiral ganglion neuron density and a highly significant increase in spiral ganglion neuron soma area compared with artificial perilymph-treated and untreated deafened cochleae. CONCLUSION: The study has demonstrated the trophic advantage of exogenous BDNF in the mature rat cochlea and provides confidence that spiral ganglion neuron rescue after sensorineural hearing loss with exogenous BDNF may have clinical application.

Animals↗

PubMed2005

Clinical application of neurotrophic factors: the potential for primary auditory neuron protection.

Sensorineural hearing loss, as a result of damage to or destruction of the sensory epithelia within the cochlea, is a common cause of deafness. The subsequent degeneration of the neural elements within the inner ear may impinge upon the efficacy of the cochlear implant. Experimental studies have demonstrated that neurotrophic factors can prevent this degeneration in animal models of deafness, and can even provide functional benefits. Neurotrophic factor therapy may therefore provide similar protective effects in humans, resulting in improved speech perception outcomes among cochlear implant patients. There are, however, numerous issues pertaining to delivery techniques and treatment regimes that need to be addressed prior to any clinical application. This review considers these issues in view of the potential therapeutic application of neurotrophic factors within the auditory system.

Animals↗

PubMed2004

Surface microstructure of the perilymphatic space: implications for cochlear implants and cell- or drug-based therapies.

OBJECTIVE: To study the surface microstructure of the scala tympani and scala vestibuli in humans and cats using scanning electron microscopy. DESIGN: Cochleas from 8 humans and 4 cats were harvested and the otic capsule and soft tissue removed before the cochleas were prepared for scanning electron microscopy. Micrographs were taken of the bony surface of both the scala tympani and scala vestibuli in each cochlear turn. The diameter and density of the micropores (canaliculi perforantes) and the thickness of the osseous spiral lamina (OSL) adjacent to Rosenthal's canal was measured. RESULTS: The human cochlea exhibits numerous canaliculi on the surface of the scala tympani, particularly associated with the OSL. There was a large range of diameters in the modiolar region of the OSL (0.2-23.0 micro m). The OSL was also very thin, with a mean thickness of 26.8 micro m in the base, tapering to 8.4 micro m in the apical turn. Far fewer canaliculi were evident in the scala vestibuli. Examination of the cat cochleas showed a similar distribution of canaliculi to that seen in the human; however, they were smaller in diameter and the OSL was thicker than in the human cochleas. CONCLUSIONS: The OSL is a thin and highly porous bony lamina that would appear to provide an open and extensive fluid communication channel between the scala tympani and Rosenthal's canal. These findings have important implications for the design and application of perimodiolar cochlear implant electrode arrays and may provide a potential route for drug- and cell-based cochlear therapies delivered via the scala tympani.

Animals↗

PubMed2004

Long-term sensorineural hearing loss induces functional changes in the rat auditory nerve.

Loss of cochlear hair cells in the rat initiates degenerative change within the primary auditory neurons (ANs) of the cochlea. These degenerative changes include loss of peripheral processes, demyelination and ultimately cell death. This pathology will affect the biophysical processes involved in action potential generation and propagation to an electrical stimulus via a cochlear implant. We measured the response properties of ANs, with particular reference to their refractory behaviour, in normal, short- (9 weeks) and long-term (> 52 weeks) deafened rats. AN loss was moderate in the short-term and severe in the long-term deafened animals. AN activity was elicited using a brief electrical stimulus delivered via a bipolar electrode array implanted into the cochlea. The general response properties of ANs recorded from deafened cochleae were similar to those observed in normal cochleae, i.e. a monotonic increase in the probability of firing and a decrease in response latency and temporal jitter with increasing stimulus intensity. However, the absolute refractory period was significantly prolonged in animals deaf for > 12 months (P = 0.0026). Deafened animals also exhibited a highly significant increase in threshold compared with normal controls (P < 0.001). These functional changes have implications for recipients of cochlear implants and potential therapies directed toward halting or reversing AN pathology.

Acoustic Stimulation↗

PubMed2003

Cochlear immunochemistry--a new technique based on gelatin embedding.

Histological processing of the cochlea for immunochemistry is often a compromise between good anatomical resolution and preservation of antigenicity. Techniques able to preserve tissue architecture invariably demand elevated temperatures and harsh chemicals or a combination of both. The likely result is reduced antigenicity, enzyme activity and nucleic acid integrity. We have modified an existing embedding medium for use in the cochlea that operates at physiological temperature and avoids denaturing agents and organic solvents. Tissue antigenicity is maximised and anatomical detail preserved, normally two mutually exclusive goals. The method is attractive because of its simplicity, speed and transparency for easy cochlear orientation. It is also likely to be adaptable for the infiltration of other heterogeneous structures prone to distortion during frozen sectioning.

Animals↗

PubMed2002

Chronic placental insufficiency has long-term effects on auditory function in the guinea pig.

Very low birth weight and growth-restricted infants have an increased risk of auditory impairments. It is uncertain whether these impairments are related to adverse pre-, peri- or postnatal events. We aimed to determine whether a period of chronic placental insufficiency (CPI) in the guinea pig results in long-term alterations to auditory function. Near mid-gestation, CPI was induced via unilateral ligation of the uterine artery. At 8 weeks of age, auditory brainstem responses (ABRs) were recorded in response to unilateral acoustic stimulation in prenatally-compromised (PC, n=8) and control animals (n=8). Stimuli consisted of 100 micros clicks, presented at 33 pulses per second (pps) and tone pip stimuli at frequencies of 2, 4, 8, 16 and 32 kHz. To examine temporal response properties, click stimuli were also presented at rates of 66, 132 and 200 pps. Normal ABR waveforms were elicited by both click and tone pip stimuli in all animals. Moreover, there was no difference between control and PC animals in stimulus detection thresholds across the frequencies examined. Using high rate click stimuli, PC animals demonstrated a significant increase in both the latency of wave III (normalised to 33 pps) and the wave I-III inter-peak interval compared to the controls. We hypothesise that these functional changes reflect alterations in myelination of the auditory brainstem and/or changes in synaptic efficacy. The results suggest subtle deficits in neural conduction in the PC guinea pig at maturity, and may have implications for speech perception abilities of low birth weight or prenatally affected infants.

Acoustic Stimulation↗

PubMed2002

A multichannel scala tympani electrode array incorporating a drug delivery system for chronic intracochlear infusion.

We have developed a novel scala tympani electrode array suitable for use in experimental animals. A unique feature of this array is its ability to chronically deliver pharmacological agents to the scala tympani. The design of the electrode array is described in detail. Experimental studies performed in guinea pigs confirm that this array can successfully deliver various drugs to the cochlea while chronically stimulating the auditory nerve.

Animals↗