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Perilymph proteomic imprint using a new tool with a nanoporous silicon chip.
Boyer E. , Berger F. , Bouamrani A. , Mombrun A. , Schmerber S.
CHU Grenoble, La Tronche, France, 2CEA LETI, Clinatec, Grenoble, France
Introduction: The physiopathology of hearing loss and intimate mechanisms of degeneration of the inner ear
structures is still largely unknown. Hence, 25 % of sensorineural hearing loss is currently unexplained. The in
vivo exploration of the cochlea is a new challenge in attempt to get new insights into the oxidative stress and
mitochondrial metabolites inducing apoptotic pathways in noise-induced hearing. Specific perilymph sampling is
impossible in vivo without contamination of cerebrospinal fluid. Due to the small size of the entrance gate of the
cochlea represented in vivo by the round window, nanotechnology aims at circumventing the technological
barrier to reach the desired intracochlear target. Our objective is to design a molecular imprint tool using nano
chips of silicon to analyze perilymph with mass spectrometry.
Material and methods: A nanoporous silicon chip (2 mm length, 300 µm width) is fixed on a medical polyether
ether ketone (PEEK) base with an inox extension cable. The device is surrounded by a retractable Teflon sheath
with a sharp distal tip to protect the silicon chip and penetrate the round window membrane.
The surgical procedure was managed on 30 WISTAR rats, 15 healthy rats and 15 rats exposed to ototoxic dose
of gentamicin (160 mg/kg/j) during 5 days. We exposed the tympanic bulla by a ventral approach and drilled the
bone capsule to have an access to the round window. Then an imprint was done, applying our tool through the
round window membrane and insert the silicon chip in cochlea. The silicon was examined in a MALDI-TOF
(Matrix-assisted laser desorption ionization - time of flight) mass spectrometry.
Results: Our preliminary results show a proteinic enrichment of the nanoporous silicon chip instead of a
standard surface of analysis. The use of a nanoporous surface allows a peptidomic and metabolomic analysis of
our perilymph sample. Imprint realization was done easily using our device. We identified specific metabolites of
perilymph and concentration gradient in our ototoxic model.
Discussion and conclusion: This new tool allows a specific perilymph protein analysis. Perspectives are the
characterization of the perilymph proteome, the discovery of hearing damage biomarkers and potential
treatments, in particular in cochlear implant surgery.