June 2023
Volume 64, Issue 8
Open Access
ARVO Annual Meeting Abstract  |   June 2023
Extracellular vesicles (EVs) from 3D cultured human bone marrow mesenchymal stem cells (hBM-MSC) demonstrated increased complexity and neurite elongation in an in-vivo corneal injury model
Author Affiliations & Notes
  • Hamed Massoumi
    Illinois Eye and Ear Infirmary, University of Illinois Chicago Department of Ophthalmology and Visual Sciences, Chicago, Illinois, United States
    University of Illinois Chicago Richard and Loan Hill Department of Biomedical Engineering, Chicago, Illinois, United States
  • Eitan Katz
    Illinois Eye and Ear Infirmary, University of Illinois Chicago Department of Ophthalmology and Visual Sciences, Chicago, Illinois, United States
  • Tara Thanh Nguyen
    Illinois Eye and Ear Infirmary, University of Illinois Chicago Department of Ophthalmology and Visual Sciences, Chicago, Illinois, United States
  • Qiang Zhou
    Illinois Eye and Ear Infirmary, University of Illinois Chicago Department of Ophthalmology and Visual Sciences, Chicago, Illinois, United States
  • Cedra Jazayerli
    Illinois Eye and Ear Infirmary, University of Illinois Chicago Department of Ophthalmology and Visual Sciences, Chicago, Illinois, United States
  • Khandaker Anwar
    Illinois Eye and Ear Infirmary, University of Illinois Chicago Department of Ophthalmology and Visual Sciences, Chicago, Illinois, United States
  • Mohammadjavad Ashraf
    Illinois Eye and Ear Infirmary, University of Illinois Chicago Department of Ophthalmology and Visual Sciences, Chicago, Illinois, United States
  • Mohammad Soleimani
    Illinois Eye and Ear Infirmary, University of Illinois Chicago Department of Ophthalmology and Visual Sciences, Chicago, Illinois, United States
  • Victor H Guaiquil
    Illinois Eye and Ear Infirmary, University of Illinois Chicago Department of Ophthalmology and Visual Sciences, Chicago, Illinois, United States
  • Mark Rosenblatt
    Illinois Eye and Ear Infirmary, University of Illinois Chicago Department of Ophthalmology and Visual Sciences, Chicago, Illinois, United States
  • Ali R Djalilian
    Illinois Eye and Ear Infirmary, University of Illinois Chicago Department of Ophthalmology and Visual Sciences, Chicago, Illinois, United States
  • Elmira Jalilian
    Illinois Eye and Ear Infirmary, University of Illinois Chicago Department of Ophthalmology and Visual Sciences, Chicago, Illinois, United States
    University of Illinois Chicago Richard and Loan Hill Department of Biomedical Engineering, Chicago, Illinois, United States
  • Footnotes
    Commercial Relationships   Hamed Massoumi None; Eitan Katz None; Tara Nguyen None; Qiang Zhou None; Cedra Jazayerli None; Khandaker Anwar None; Mohammadjavad Ashraf None; Mohammad Soleimani None; Victor Guaiquil None; Mark Rosenblatt None; Ali Djalilian None; Elmira Jalilian None
  • Footnotes
    Support  This work was supported by National Center for Advancing Translational Sciences, National Institutes of Health, under Grant KL2TR002002, R01 EY024349 (ARD), Core Grant for Vision Research EY01792 (MIR) from NEI/NIH; Research to Prevent Blindness Unrestricted Departmental Grant, Physician-Scientist Award both from Research to Prevent Blindness & UG3/UH3 EY031809 from NEI, NIH Grant P30EY001792,
Investigative Ophthalmology & Visual Science June 2023, Vol.64, 2350. doi:
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      Hamed Massoumi, Eitan Katz, Tara Thanh Nguyen, Qiang Zhou, Cedra Jazayerli, Khandaker Anwar, Mohammadjavad Ashraf, Mohammad Soleimani, Victor H Guaiquil, Mark Rosenblatt, Ali R Djalilian, Elmira Jalilian; Extracellular vesicles (EVs) from 3D cultured human bone marrow mesenchymal stem cells (hBM-MSC) demonstrated increased complexity and neurite elongation in an in-vivo corneal injury model. Invest. Ophthalmol. Vis. Sci. 2023;64(8):2350.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose : Corneal nerve damage leads to painful ocular surface sensitivity, decreased tear production, and serious vision problems. Treatment options such as eye drops are not effective and often fail to manage symptoms in severe cases. Therefore, there is an urgent clinical need to develop therapies to promote rapid corneal nerve regeneration. We showed that the in vitro usage of EVs obtained from hBM-MSCs can promote nerve regeneration. In this study, we evaluated the in vivo effect of EVs on nerve regeneration in wounded cornea models in mice.

Methods : The 2D monolayer flasks and 3D bioreactors were used to culture hBM-MSCs. Ultracentrifugation was used to isolate EVs. The concentration and size distribution of the EVs were characterized by NanoSight. A 2 mm corneal debridement was performed on C57 mice, followed by immunostaining and slit-lamp imaging. At days 0, 2, and 6, a 25 μL subconjunctival injection of PBS, 2D- and 3D-derived EVs was performed on the control and the two treatment groups. Wound healing was monitored for 48 h. Animals were euthanized after 12 days and corneas were dissected from their enucleated eyeballs. Whole-mount β3-tubulin nerve staining was performed followed by confocal microscopy to evaluate nerve regeneration. Images were analyzed using Neurolucida software.

Results : NanoSight analysis revealed a 28-fold increase in EV production using a 3D culture system vs. a 2D monolayer. However, the 3D and 2D-EV size distributions of 139 nm and 146 nm, were approximately the same. Wound closure after 24 h was 91.7% and 89.9% for samples treated with 3D- and 2D-EVs, respectively, whereas the control group showed 88.2% wound closure. Complete wound closure for all samples occurred at 48 h. Neurolucida analysis revealed a significant increase in nerve regeneration using 3D-EV treatment with 40.62 ± 26.8 mm neurite length compared to 2D-EV treatment with 22.36 ± 8.7 mm and the control group with 20.98 ± 2.9 mm.

Conclusions : These results demonstrated that the 3D-derived EVs can significantly increase nerve regeneration and elongation in in-vivo models of corneal injury compared to 2D-derived EVs. Further studies will reveal the molecular mechanisms of 3D- and 2D-EVs in nerve regeneration. These studies will enable us to develop efficient therapeutics for the restoration of damaged corneal nerves.

This abstract was presented at the 2023 ARVO Annual Meeting, held in New Orleans, LA, April 23-27, 2023.

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