Betsy Szeto1, Aykut Aksit2, Chris Valentini1, Michelle Yu1, Emily G Werth3, Shahar Goeta3, Chuanning Tang3, Lewis M Brown3, Elizabeth S Olson4, Jeffrey W Kysar5, Anil K Lalwani6. 1. Department of Otolaryngology - Head and Neck Surgery, Columbia University Vagelos College of Physicians and Surgeons, 180 Fort Washington Avenue, Harkness Pavilion, 8th Floor, New York, NY 10032, United States. 2. Department of Mechanical Engineering, Columbia University, New York, NY, United States. 3. Quantitative Proteomics and Metabolomics Center, Department of Biological Sciences, Columbia University, New York, NY, United States. 4. Department of Biomedical Engineering, Columbia University, New York, NY, United States; Department of Otolaryngology - Head and Neck Surgery, Columbia University Vagelos College of Physicians and Surgeons, 180 Fort Washington Avenue, Harkness Pavilion, 8th Floor, New York, NY 10032, United States. 5. Department of Mechanical Engineering, Columbia University, New York, NY, United States; Department of Otolaryngology - Head and Neck Surgery, Columbia University Vagelos College of Physicians and Surgeons, 180 Fort Washington Avenue, Harkness Pavilion, 8th Floor, New York, NY 10032, United States. 6. Department of Otolaryngology - Head and Neck Surgery, Columbia University Vagelos College of Physicians and Surgeons, 180 Fort Washington Avenue, Harkness Pavilion, 8th Floor, New York, NY 10032, United States; Department of Mechanical Engineering, Columbia University, New York, NY, United States. Electronic address: anil.lalwani@columbia.edu.
Abstract
BACKGROUND: Inner ear diagnostics is limited by the inability to atraumatically obtain samples of inner ear fluid. The round window membrane (RWM) is an attractive portal for accessing perilymph samples as it has been shown to heal within one week after the introduction of microperforations. A 1 µL volume of perilymph is adequate for proteome analysis, yet the total volume of perilymph within the scala tympani of the guinea pig is limited to less than 5 µL. This study investigates the safety and reliability of a novel hollow microneedle device to aspirate perilymph samples adequate for proteomic analysis. METHODS: The guinea pig RWM was accessed via a postauricular surgical approach. 3D-printed hollow microneedles with an outer diameter of 100 µm and an inner diameter of 35 µm were used to perforate the RWM and aspirate 1 µL of perilymph. Two perilymph samples were analyzed by liquid chromatography-mass spectrometry-based quantitative proteomics as part of a preliminary study. Hearing was assessed before and after aspiration using compound action potential (CAP) and distortion product otoacoustic emissions (DPOAE). RWMs were harvested 72 h after aspiration and evaluated for healing using confocal microscopy. RESULTS: There was no permanent damage to hearing at 72 h after perforation as assessed by CAP (n = 7) and DPOAE (n = 8), and all perforations healed completely within 72 h (n = 8). In the two samples of perilymph analyzed, 620 proteins were detected, including the inner ear protein cochlin, widely recognized as a perilymph marker. CONCLUSION: Hollow microneedles can facilitate aspiration of perilymph across the RWM at a quality and volume adequate for proteomic analysis without causing permanent anatomic or physiologic dysfunction. Microneedles can mediate safe and effective intracochlear sampling and show great promise for inner ear diagnostics.
BACKGROUND: Inner ear diagnostics is limited by the inability to atraumatically obtain samples of inner ear fluid. The round window membrane (RWM) is an attractive portal for accessing perilymph samples as it has been shown to heal within one week after the introduction of microperforations. A 1 µL volume of perilymph is adequate for proteome analysis, yet the total volume of perilymph within the scala tympani of the guinea pig is limited to less than 5 µL. This study investigates the safety and reliability of a novel hollow microneedle device to aspirate perilymph samples adequate for proteomic analysis. METHODS: The guinea pig RWM was accessed via a postauricular surgical approach. 3D-printed hollow microneedles with an outer diameter of 100 µm and an inner diameter of 35 µm were used to perforate the RWM and aspirate 1 µL of perilymph. Two perilymph samples were analyzed by liquid chromatography-mass spectrometry-based quantitative proteomics as part of a preliminary study. Hearing was assessed before and after aspiration using compound action potential (CAP) and distortion product otoacoustic emissions (DPOAE). RWMs were harvested 72 h after aspiration and evaluated for healing using confocal microscopy. RESULTS: There was no permanent damage to hearing at 72 h after perforation as assessed by CAP (n = 7) and DPOAE (n = 8), and all perforations healed completely within 72 h (n = 8). In the two samples of perilymph analyzed, 620 proteins were detected, including the inner ear protein cochlin, widely recognized as a perilymph marker. CONCLUSION: Hollow microneedles can facilitate aspiration of perilymph across the RWM at a quality and volume adequate for proteomic analysis without causing permanent anatomic or physiologic dysfunction. Microneedles can mediate safe and effective intracochlear sampling and show great promise for inner ear diagnostics.
Authors: A Santimetaneedol; Z Wang; D N Arteaga; A Aksit; C Prevoteau; M Yu; H Chiang; D Fafalis; A K Lalwani; J W Kysar Journal: Colloids Surf B Biointerfaces Date: 2019-07-04 Impact factor: 5.268
Authors: Jonathan C Palmer; Megan S Lord; Jeremy L Pinyon; Andrew K Wise; Nigel H Lovell; Paul M Carter; Ya Lang Enke; Gary D Housley; Rylie A Green Journal: Laryngoscope Date: 2017-11-02 Impact factor: 3.325
Authors: Miguel Arriaga; Daniel N Arteaga; Dimitrios Fafalis; Michelle Yu; Xun Wang; Karen E Kasza; Anil K Lalwani; Jeffrey W Kysar Journal: Acta Biomater Date: 2021-09-24 Impact factor: 8.947
Authors: Betsy Szeto; Chris Valentini; Aykut Aksit; Emily G Werth; Shahar Goeta; Lewis M Brown; Elizabeth S Olson; Jeffrey W Kysar; Anil K Lalwani Journal: J Proteome Res Date: 2021-07-22 Impact factor: 4.466