Significance: Worldwide, ∼460 million people suffer from disabling hearing impairment. Many of these patients are still not sufficiently supplied with currently available auditory technologies. Optical stimulation of the hearing organ offers a promising alternative for a new generation of auditory prostheses.
Aim: To assess the biocompatibility margins of our laser pulse amplitude strategy in vitro, we designed a protocol and present the effects on normal human dermal fibroblasts, human chondrocytes, and human osteoblasts.
Approach: Laser pulses of 532 nm were applied over 120 s using our laser pulse amplitude modulation strategy. We then assessed cell viability and cytotoxicity through fluorescence staining and quantitative polymerase chain reaction-analysis regarding 84 key player-genes for cytotoxicity and stress response.
Results: The first in vitro biocompatibility margins for our stimulation parameters applied to cells of the peripheral hearing organ were between 200 and 223 mW (3348 J/cm2). After irradiation with a subphototoxic laser power of 199 mW (2988 J/cm2), only the fibroblasts showed a significant upregulation of GADD45G.
Conclusion: Further studies are underway to optimize parameters for the optoacoustic stimulation of the auditory system. Our protocol and results on laser–tissue interactions can be useful for translational laser applications in various other irradiated biological tissues.
Hearing impairment affects ∼460 million people worldwide. Conservative therapies, such as hearing aids, bone conduction systems, and middle ear implants, do not always sufficiently compensate for this deficit. The optical stimulation is currently under investigation as an alternative stimulation strategy for the activation of the hearing system. To assess the biocompatibility margins of this emerging technology, we established a method applicable in whole-mount preparations of murine tympanic membranes (TM). We irradiated the TM of anesthetized mice with 532-nm laser pulses at an average power of 50, 89, 99, and 125 mW at two different locations of the TM and monitored the hearing function with auditory brainstem responses. Laser-power-dependent negative side effects to the TM were observed at power levels exceeding 89 mW. Although we did not find any significant negative effects of optical stimulation on the hearing function in these mice, based on the histology results further studies are necessary for optimization of the used parameters.
The tympanic membrane (TM) separates the outer ear from the tympanic cavity. Repeated pathologies can permanently decrease its tension, inducing conductive hearing loss and adhesive processes up to cholesteatoma. The current main therapy is its surgical reconstruction. Even though lasers have been proposed to tighten atrophic TMs, details of this effect, specifically histological analyses, are missing. We therefore used laser pulses to induce TM collagen remodeling in an animal model to compare the histological and electrophysiological effects of different applied laser intensities before entering clinical studies. We irradiated Fuchsin-stained areas of the TM in anesthetized mice with 532-nm laser-pulses of 10 mW for 30 s (0.3 J), 25 mW for 30 s (0.75 J) or 50 mW for 30 s (1.5 J) monitoring hearing with auditory brainstem responses (ABRs). The mice were sacrificed after 2 to 8 weeks and histologically analyzed. An increase in the TM thickness within the defined, stained, and irradiated areas could be observed after 4 weeks. Polarized light microscopy and transmission electron microscopy demonstrated the tissue volume increase majorly due to new collagen-fibrils. Directly after irradiation, ABR thresholds did not increase. We herein demonstrate a controlled laser-induced collagen remodeling within defined areas of the TM. This method might be the prophylactic solution for chronic inflammatory ear pathologies related to decreased TM tension.
The interaction between biological tissues and light of a certain wavelength is influenced by the optical properties of each tissue. These are typically estimated from of the measurements of the main characteristics of the radiations pathway.
We herein present our approach that uses a customized version of the Monte Carlo Multi-Layer Algorithm (MCML)1 to simulate the radiation propagation through biological tissues. We assumed a set of optical properties for each tissue and simulated the above mentioned measurements in silico. A comparison was then done between the results of the simulation and the results of real measurements.2 Further, an optimization algorithm searched the set of optical properties that best fit the real optical properties of each tissue. This algorithm was based on adaptions of the Monte-Carlo Simulated Annealing algorithm3 and the Downhill Simplex algorithm4 We implemented the MCML using NVidias CUDA application programming interface to speed up the optimization procedure. We validated the software by using van de Hulsts table for Henyey-Greenstein scattering.2 A linear regression resulted in coefficients of determination between 0.929 and 0.973 for the optical properties. Our results prove that our algorithm can be effectively used for the determination of the optical properties of turbid media.
One first application for this software is the support in the development of a new generation of hearing devices based on optical energy.
Repeated pathologies of the tympanic membrane (TM) decrease its tension inducing conductive hearing loss and adhesive processes up to cholesteatoma. Our results regarding the development of a laser based noninvasive procedure to strengthen the structure of the TM are herein presented.
The hearing performance with conventional hearing aids and cochlear implants is dramatically reduced in noisy environments and for sounds more complex than speech (e. g. music), partially due to the lack of localized sensorineural activation across different frequency regions with these devices. Laser light can be focused in a controlled manner and may provide more localized activation of the inner ear, the cochlea. We sought to assess whether visible light with parameters that could induce an optoacoustic effect (532 nm, 10-ns pulses) would activate the cochlea. Auditory brainstem responses (ABRs) were recorded preoperatively in anesthetized guinea pigs to confirm normal hearing. After opening the bulla, a 50-µm core-diameter optical fiber was positioned in the round window niche and directed toward the basilar membrane. Optically induced ABRs (OABRs), similar in shape to those of acoustic stimulation, were elicited with single pulses. The OABR peaks increased with energy level (0.6 to 23 µJ/pulse) and remained consistent even after 30 minutes of continuous stimulation at 13 µJ, indicating minimal or no stimulation-induced damage within the cochlea. Our findings demonstrate that visible light can effectively and reliably activate the cochlea without any apparent damage. Further studies are in progress to investigate the frequency-specific nature and mechanism of green light cochlear activation.
The success of conventional hearing aids and electrical cochlear implants have generally been limited to hearing in quiet situations, in part due to a lack of localized (i.e., frequency specificity) sensorineural activation and subsequent impaired speech discrimination in noise. Laser light is a source of energy that can be focused in a controlled manner and may provide more localized activation of the inner ear, the cochlea. Compound action potentials have been elicited using 2.12 µm laser pulses through activation of auditory nerve fibers (Izzo et al. 2006). Laser stimulation (813 nm) of the cochlea has shown to induce basilar membrane motion and cochlear microphonic potentials (Fridberger et al. 2006). We sought to assess if visible light (green, 532 nm, 10 ns pulses) could be used to consistently activate the cochlea. The laser parameters were selected based on our initial attempt to induce an optoacoustic effect as the energy transfer mechanism to the cochlea. Click evoked auditory brainstem responses (AABRs) were recorded preoperatively in ketamine-anesthetized guinea pigs to confirm normal hearing. The bulla and then the cochlea were exposed. Optically evoked ABRs (OABR) were recorded in response to laser stimulation with a 50 µm optical fiber (532 nm, 10 ns pulses, 500 repetitions, 10 pulses/s; Nd:YAG laser) at the round window (RW) directed towards the basilar membrane (BM).
OABRs similar in morphology to acoustically evoked ABRs, except for shorter latencies, were obtained for stimulation through the RW with energy levels between 1.7-30 µJ/pulse. The OABRs increased with increasing energy level reaching a saturation level around 13-15 µJ/pulse. Furthermore the responses remained consistent across stimulation over time, including stimulation at 13 µJ/pulse for over 30 minutes, indicating minimal or no damage within the cochlea with this type of laser stimulation.
Overall we have demonstrated that laser light stimulation with 532 nm has potential for a new type of auditory prosthesis that can activate the cochlea without any apparent functional damage. Further studies are needed to determine the optimal laser parameters and fiber placement locations for localized and tonotopic activation.
The cochlea is the mammalian organ of hearing. Its predominant vibratory element, the basilar membrane, is tonotopically tuned, based on the spatial variation of its mass and stiffness. The constituent collagen fibers of the basilar membrane affect its stiffness. Laser irradiation can induce collagen remodeling and deposition in various tissues. We tested whether similar effects could be induced within the basilar membrane. Trypan blue was perfused into the scala tympani of anesthetized mice to stain the basilar membrane. We then irradiated the cochleas with a 694-nm pulsed ruby laser at 15 or 180 J/cm2. The mice were sacrificed 14 to 16 days later and collagen organization was studied. Polarization microscopy revealed that laser irradiation increased the birefringence within the basilar membrane in a dose-dependent manner. Electron microscopy demonstrated an increase in the density of collagen fibers and the deposition of new fibrils between collagen fibers after laser irradiation. As an assessment of hearing, auditory brainstem response (ABR) thresholds were found to increase moderately after 15 J/cm2 and substantially after 180 J/cm2. Our results demonstrate that collagen remodeling and new collagen deposition occurs within the basilar membrane after laser irradiation in a similar fashion to that found in other tissues.