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Preliminary Characterization of Extracellular Vesicles From Auditory HEI-OC1 Cells.

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Title: Preliminary Characterization of Extracellular Vesicles From Auditory HEI-OC1 Cells.
Authors: Kalinec, Gilda M., Cohn, Whitaker, Whitelegge, Julian P., Faull, Kym F., Kalinec, Federico
Source: Annals of Otology, Rhinology & Laryngology. 2019 Supplement, Vol. 128, p52S-60S. 9p.
Subjects: Cochlea, Drug delivery systems, Extracellular space, Hair cells, Inner ear, Liquid chromatography, Mass spectrometry, Nanoparticles, Proteomics, Dexamethasone
Abstract: Objectives: Isolate, purify, and characterize extracellular vesicles (EVs) obtained from auditory HEI-OC1 cells, and evaluate their suitability for intracochlear transport and delivery of pharmacological drugs and/or pro-resolution mediators of acute inflammatory processes. Methods: HEI-OC1 EVs were isolated and purified using the exoEasy Maxi Kit, and their size was evaluated by nanoparticle tracking techniques. Bottom-up proteomics of the EVs, either freshly obtained or stored for up to 4 months at -20°C, was performed by LC-ESI-MS/MS. LC-ESI-MS/MS-MRM was used to measure the loading of dexamethasone inside EVs following co-incubation at room temperature for 1 hour with and without 5 minutes sonication. Results: Routinely, we were able to obtain purified fractions of >2 x 109 EVs/mL, with diameters varying between 50 and 800 nm. Bottom-up proteomics showed that among the most abundant EVs proteins, 19.2% were cytoplasmic, 17.2% were membrane localized, 12.3% were cytosolic, and 14.6% were nucleolar. No significant differences between fresh and stored EVs were detected. Importantly, co-incubation of HEI-OC1 EVs (1 x 108 EVs/mL) with dexamethasone (10 mM) resulted in the incorporation of 10.1 ± 1.9 nM dexamethasone per milliliter of EVs suspension. Conclusions: Altogether, the results suggest that EVs from HEI-OC1 cells could be advantageously used as biological nanocarriers for the delivery of specific molecules and pharmacological drugs into the inner ear. [ABSTRACT FROM AUTHOR]
Copyright of Annals of Otology, Rhinology & Laryngology is the property of Sage Publications Inc. and its content may not be copied or emailed to multiple sites without the copyright holder's express written permission. Additionally, content may not be used with any artificial intelligence tools or machine learning technologies. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)
Database: Psychology and Behavioral Sciences Collection
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  Data: Preliminary Characterization of Extracellular Vesicles From Auditory HEI-OC1 Cells.
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  Data: <searchLink fieldCode="AR" term="%22Kalinec%2C+Gilda+M%2E%22">Kalinec, Gilda M.</searchLink><br /><searchLink fieldCode="AR" term="%22Cohn%2C+Whitaker%22">Cohn, Whitaker</searchLink><br /><searchLink fieldCode="AR" term="%22Whitelegge%2C+Julian+P%2E%22">Whitelegge, Julian P.</searchLink><br /><searchLink fieldCode="AR" term="%22Faull%2C+Kym+F%2E%22">Faull, Kym F.</searchLink><br /><searchLink fieldCode="AR" term="%22Kalinec%2C+Federico%22">Kalinec, Federico</searchLink>
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  Data: <searchLink fieldCode="JN" term="%22Annals+of+Otology%2C+Rhinology+%26+Laryngology%22">Annals of Otology, Rhinology & Laryngology</searchLink>. 2019 Supplement, Vol. 128, p52S-60S. 9p.
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  Data: <searchLink fieldCode="DE" term="%22Cochlea%22">Cochlea</searchLink><br /><searchLink fieldCode="DE" term="%22Drug+delivery+systems%22">Drug delivery systems</searchLink><br /><searchLink fieldCode="DE" term="%22Extracellular+space%22">Extracellular space</searchLink><br /><searchLink fieldCode="DE" term="%22Hair+cells%22">Hair cells</searchLink><br /><searchLink fieldCode="DE" term="%22Inner+ear%22">Inner ear</searchLink><br /><searchLink fieldCode="DE" term="%22Liquid+chromatography%22">Liquid chromatography</searchLink><br /><searchLink fieldCode="DE" term="%22Mass+spectrometry%22">Mass spectrometry</searchLink><br /><searchLink fieldCode="DE" term="%22Nanoparticles%22">Nanoparticles</searchLink><br /><searchLink fieldCode="DE" term="%22Proteomics%22">Proteomics</searchLink><br /><searchLink fieldCode="DE" term="%22Dexamethasone%22">Dexamethasone</searchLink>
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  Label: Abstract
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  Data: Objectives: Isolate, purify, and characterize extracellular vesicles (EVs) obtained from auditory HEI-OC1 cells, and evaluate their suitability for intracochlear transport and delivery of pharmacological drugs and/or pro-resolution mediators of acute inflammatory processes. Methods: HEI-OC1 EVs were isolated and purified using the exoEasy Maxi Kit, and their size was evaluated by nanoparticle tracking techniques. Bottom-up proteomics of the EVs, either freshly obtained or stored for up to 4 months at -20°C, was performed by LC-ESI-MS/MS. LC-ESI-MS/MS-MRM was used to measure the loading of dexamethasone inside EVs following co-incubation at room temperature for 1 hour with and without 5 minutes sonication. Results: Routinely, we were able to obtain purified fractions of >2 x 109 EVs/mL, with diameters varying between 50 and 800 nm. Bottom-up proteomics showed that among the most abundant EVs proteins, 19.2% were cytoplasmic, 17.2% were membrane localized, 12.3% were cytosolic, and 14.6% were nucleolar. No significant differences between fresh and stored EVs were detected. Importantly, co-incubation of HEI-OC1 EVs (1 x 108 EVs/mL) with dexamethasone (10 mM) resulted in the incorporation of 10.1 ± 1.9 nM dexamethasone per milliliter of EVs suspension. Conclusions: Altogether, the results suggest that EVs from HEI-OC1 cells could be advantageously used as biological nanocarriers for the delivery of specific molecules and pharmacological drugs into the inner ear. [ABSTRACT FROM AUTHOR]
– Name: AbstractSuppliedCopyright
  Label:
  Group: Ab
  Data: <i>Copyright of Annals of Otology, Rhinology & Laryngology is the property of Sage Publications Inc. and its content may not be copied or emailed to multiple sites without the copyright holder's express written permission. Additionally, content may not be used with any artificial intelligence tools or machine learning technologies. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract.</i> (Copyright applies to all Abstracts.)
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        Value: 10.1177/0003489419836226
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        Text: English
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        PageCount: 9
        StartPage: 52S
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      – SubjectFull: Cochlea
        Type: general
      – SubjectFull: Drug delivery systems
        Type: general
      – SubjectFull: Extracellular space
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      – SubjectFull: Hair cells
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      – SubjectFull: Inner ear
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      – SubjectFull: Liquid chromatography
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      – SubjectFull: Mass spectrometry
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      – SubjectFull: Nanoparticles
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      – SubjectFull: Proteomics
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      – SubjectFull: Dexamethasone
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      – TitleFull: Preliminary Characterization of Extracellular Vesicles From Auditory HEI-OC1 Cells.
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              Text: 2019 Supplement
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              Y: 2019
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              Value: 128
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