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    Biomechanics

    (D-148) The Design and Development of a Phonomimetic Bioreactor

    Friday, October 13, 2023
    9:30 AM – 10:30 AM PDT
    Location: Exhibit Hall - Row D - Poster # 148
    Introduction::

    Vocal fold disorders are prevalent communication disorders that can affect individuals of all ages and genders. The leading cause of these disorders is the misuse and abuse of the vocal folds leading to different clinical disorders such as nodules or polyps (1). The purpose of this design is to develop a phonomimetic bioreactor capable of mimicking human acoustical patterns, resembling the effects caused by the misuse and abuse of the vocal folds. This design aims to facilitate the investigation of tissue integration responses when subjected to acoustical stimulation.



    Materials and Methods:: The housing shell of the phonomimetic bioreactor was 3D printed in three separate parts using polylactic acid. A DA175-8 aluminum cone woofer (Dayton Audio, Springboro, OH) was inserted in the lower housing section of the phonomimetic bioreactor (Fig 1.C and Fig 1.D). This driver was selected due to its capability to project frequencies within the human vocal range for speech (60-180 Hz for males, 160-300 Hz for females) (2). The bottom of the regular six-well plates was removed prior to insertion and replaced with cured Sylgard 184 elastomer that was then coated with collagen type IV neutralized with NaOH. The efficacy of the sound transmission through the Silicone-collagen based well plates was evaluated using a Laser Doppler Vibrometer. As a preliminary test for cell culture integrity, HeLa cells were cultured and seeded on the collagen-coated Sylgard membrane and cultured for three days to reach 90% confluency. The cells were then stimulated at 170 Hz for 3 hours (2) in the incubator (5% CO2 and 37ºC), then samples (n=3) were fixed in 4% Paraformaldehyde for further staining (Fig 1. B). To visualize, the cells were stained with Crystal violet and further imaged. All chemicals were purchased from Fisher Scientific (Hampton, NH).

    Results, Conclusions, and Discussions:: Figure 1 exhibits the bioreactor preliminary design and assembly (Fig 1.C). The efficacy of the assembled bioreactor tested with Laser Doppler Vibrometer showed minimized frequency shift and 42% decrease in amplitude. (Fig 1.A). The extended excitation of the cells at the consistent frequency of 170 Hz over the course of 3 hours produced changes in the collagen gel integrity and also cell attachment. This was represented as the large gaps observed within the cell colonies, which was indicative of disruption as a direct result of the stimulation (Fig 1.B).
    The design and testing of a phonomimetic bioreactor were conducted to ensure its accuracy and similarity in replicating the frequency and amplitudes found within the human voice range. An in vitro model was employed, using HeLa cells, to study the effects of acoustical stimulation on cell culture integrity. Further studies are required to understand the impact of voice characteristics on cell culture integrity.



    Acknowledgements (Optional): :

    We would like to acknowledge the support from the College of Engineering, Technology, and Architecture through the Faculty-Student Engagement Grant at the University of Hartford.



    References (Optional): :

    < !1.      Naqvi Y. et al. "Functional voice disorders." (2020).


    < !2.Lortie C., et al. "Effects of age on the amplitude, frequency and perceived quality of voice." Age 37 (2015): 1-24.