Multimodal additive manufacturing of biomimetic tympanic membrane replacements with near tissue-like acousto-mechanical and biological properties.

The three additive manufacturing techniques fused deposition modeling, gel plotting and melt electrowriting were combined to develop a mimicry of the tympanic membrane (TM) to tackle large TM perforations caused by chronic otitis media. The mimicry of the collagen fiber orientation of the TM was accompanied by a study of multiple funnel-shaped mimics of the TM morphology, resulting in mechanical and acoustic properties similar to those of the eardrum. For the different 3D printing techniques used, the process parameters were optimized to allow reasonable microfiber arrangements within the melt electrowriting setup. Interestingly, the fiber pattern was less important for the acousto-mechanical properties than the overall morphology. Furthermore, the behavior of keratinocytes and fibroblasts is crucial for the repair of the TM, and an in vitro study showed a high biocompatibility of both primary cell types while mimicking the respective cell layers of the TM. A simulation of the in vivo ingrowth of both cell types resulted in a cell growth orientation similar to the original collagen fiber orientation of the TM. Overall, the combined approach showed all the necessary parameters to support the growth of a neo-epithelial layer with a similar structure and morphology to the original membrane. It therefore offers a suitable alternative to autologous materials for the treatment of chronic otitis media. STATEMENT OF SIGNIFICANCE: Millions of people worldwide suffer from chronic middle ear infections. Although the tympanic membrane (TM) can be reconstructed with autologous materials, the grafts used for this purpose require extensive manual preparation during surgery. This affects not only the hearing ability but also the stability of the reconstructed TM, especially in the case of full TM reconstruction. The synthetic alternative presented here mimicked not only the fibrous structure of the TM but also its morphology, resulting in similar acousto-mechanical properties. Furthermore, its high biocompatibility supported the migration of keratinocytes and fibroblasts to form a neo-epithelial layer. Overall, this completely new TM replacement was achieved by combining three different additive manufacturing processes.

Mimicking the Human Tympanic Membrane: The Significance of Scaffold Geometry.

The human tympanic membrane (TM) captures sound waves from the environment and transforms them into mechanical motion. The successful transmission of these acoustic vibrations is attributed to the unique architecture of the TM. However, a limited knowledge is available on the contribution of its discrete anatomical features, which is important for fabricating functional TM replacements. This work synergizes theoretical and experimental approaches toward understanding the significance of geometry in tissue-engineered TM scaffolds. Three test designs along with a plain control are chosen to decouple some of the dominant structural elements, such as the radial and circumferential alignment of the collagen fibrils. In silico models suggest a geometrical dependency of their mechanical and acoustical responses, where the presence of radially aligned fibers is observed to have a more prominent effect compared to their circumferential counterparts. Following which, a hybrid fabrication strategy combining electrospinning and additive manufacturing has been optimized to manufacture biomimetic scaffolds within the dimensions of the native TM. The experimental characterizations conducted using macroindentation and laser Doppler vibrometry corroborate the computational findings. Finally, biological studies with human dermal fibroblasts and human mesenchymal stromal cells reveal a favorable influence of scaffold hierarchy on cellular alignment and subsequent collagen deposition.

Biomimetic Tympanic Membrane Replacement Made by Melt Electrowriting.

The tympanic membrane (TM) transfers sound waves from the air into mechanical motion for the ossicular chain. This requires a high sensitivity to small dynamic pressure changes and resistance to large quasi-static pressure differences. The TM achieves this by providing a layered structure of about 100µm in thickness, a low flexural stiffness, and a high tensile strength. Chronically infected middle ears require reconstruction of a large area of the TM. However, current clinical treatment can cause a reduction in hearing. With the novel additive manufacturing technique of melt electrowriting (MEW), it is for the first time possible to fabricate highly organized and biodegradable membranes within the dimensions of the TM. Scaffold designs of various fiber composition are analyzed mechanically and acoustically. It can be demonstrated that by customizing fiber orientation, fiber diameter, and number of layers the desired properties of the TM can be met. An applied thin collagen layer seals the micropores of the MEW-printed membrane while keeping the favorable mechanical and acoustical characteristics. The determined properties are beneficial for implantation, closely match those of the human TM, and support the growth of a neo-epithelial layer. This proves the possibilities to create a biomimimetic TM replacement using MEW.

Static and dynamic forces in the incudostapedial joint gap.

Dynamic pressure at the tympanic membrane is transformed and subsequently transferred through the ossicular chain in the form of forces and moments. The forces are primarily transferred to the inner ear. They are transferred partly to the stapedial annular ligament which exhibits non-linear behavior and stiffens for larger static forces. In unventilated middle ears, static pressure is additionally transferred to the ossicles. The purpose of this study was to measure the force inside the ossicular chain as a physiological parameter. We determined the forces which act for dynamic sound transmission and for static load on the ossicular chain. The study is the first one which introduces these forces. The static forces have direct impact on clinically relevant questions for middle ear reconstructions with passive or active prosthesis. The dynamic forces have an impact on the development of middle ear sensors. Quasi-static forces in the incudostapedial joint (ISJ) gap were measured with two different sensor types in 17 temporal bones. The sensing elements, a single crystal piezo and a strain gauge element for validation, were bonded to a thin flexible titanium plate and encapsulated in a titanium housing to allow the acquisition of the applied force signal inside the ossicular chain. Dynamic forces were measured in 11 temporal bones with the piezo sensor. We measured a static force of 23 mN in the ISJ after sensor insertion. The mean force for dynamic physiological acoustic excitation from 250 Hz to 6 kHz was 26 µN/Pa. If the tympanic membrane is loaded with a static pressure, the static force in the ISJ increases up to 1 N for a maximum static pressure load scenario of 30 kPa.

Function, Applicability, and Properties of a Novel Flexible Total Ossicular Replacement Prosthesis With a Silicone Coated Ball and Socket Joint.

HYPOTHESIS: A total ossicular replacement prosthesis (TORP) with a silicone coated ball and socket joint (BSJ) is able to compensate pressure changes and therefore provide better sound transmission compared with rigid prostheses. BACKGROUND: Dislocation and extrusion are known complications after TORP reconstruction, leading to revisions and recurrent hearing loss. Poor aeration of the middle ear, scar tension, and static pressure variations in conjunction with rigid prosthesis design causes high tension at the implant coupling points. METHODS: A novel TORP prototype with a silicone coated BSJ has been developed. Experimental measurements were performed on nine fresh cadaveric human temporal bones of which five were used for a comparison between rigid TORP and flexible TORP tympanoplasty. The middle ear transfer function was measured at ambient pressure and at 2.5 kPa, both positive and negative pressure, applied in the ear canal. RESULTS: The flexible TORP design yields a better transmission of sound after implantation and at negative pressure inside the tympanic cavity, compared with rigid TORP. In average, it provides an equivalent sound transfer like the intact middle ear. At positive pressure, the flexible TORP performs slightly worse. Both performed worse than the intact middle ear, which is related to an uplifting of the prostheses. CONCLUSION: The findings may be considered preliminary as this experimental study was limited to just one of the many different possible situations of tympanoplasty and it involved a small sample size. Nevertheless, the results with the flexible TORP were promising and could encourage further investigations on such prostheses.

Fully implantable hearing aid in the incudostapedial joint gap.

A fully implantable hearing aid is introduced which is a combined sensor-actuator-transducer designed for insertion into the incudostapedial joint gap (ISJ). The active elements each consist of a thin titanium membrane with an applied piezoelectric single crystal. The effectiveness of the operating principle is verified in a temporal bone study. We also take a closer look at the influence of an implantation-induced increase in middle ear stiffness on the transducer's output. An assembly of the transducer with 1 mm thickness is built and inserted into six temporal bones. At this thickness, the stiffness of the annular ligament is considerably increased, which leads to a loss in functional gain for the transducer. It is assumed that a thinner transducer would reduce this effect. In order to examine the performance for a prospective reduced pretension, we increased the gap size at the ISJ by 0.5 mm by removing the capitulum of the stapes in four temporal bones. The TM is stimulated with a broadband multisine sound signal in the audiological frequency range. The movement of the stapes footplate is measured with a laser Doppler vibrometer. The sensor signal is digitally processed and the amplified signal drives the actuator. The resulting feedback is minimized by an active noise control least mean square (LMS) algorithm which is implemented on a field programmable gate array. The dynamic range and the functional gain of the transducer in the temporal bones are determined. The results are compared to measurements from temporal bones without ISJ extension and to the results of Finite Elements Model (FE model) simulations. In the frequency range above 2 kHz a functional gain of 30 dB and more is achieved. This proposes the transducer as a potential treatment for high frequency hearing loss, e.g. for patients with noise-induced hearing loss. The transducer offers sufficient results for a comprehensive application. Adaptations in the transducer design or surgical approach are necessary to cope with ligament stiffening issues. These cause insufficient performance for low frequencies under 1 kHz.