June 2020
Volume 61, Issue 7
Free
ARVO Annual Meeting Abstract  |   June 2020
Production of Bio-active Nanoparticle from Human Corneal Extracellular Matrix
Author Affiliations & Notes
  • Ghasem Yazdanpanah
    Illinois Eye and Ear Infirmary, UIC, Chicago, Illinois, United States
  • Meisam Omidi
    School of Dentistry, Marquette University, Milwaukee, Wisconsin, United States
  • Ritu Shah
    Illinois Eye and Ear Infirmary, UIC, Chicago, Illinois, United States
  • Sayena Jabbehdari
    Illinois Eye and Ear Infirmary, UIC, Chicago, Illinois, United States
  • Khandaker N Anwar
    Illinois Eye and Ear Infirmary, UIC, Chicago, Illinois, United States
  • Xiang Shen
    Illinois Eye and Ear Infirmary, UIC, Chicago, Illinois, United States
  • Behnam Rabiee
    Illinois Eye and Ear Infirmary, UIC, Chicago, Illinois, United States
  • Ilham Putra
    Illinois Eye and Ear Infirmary, UIC, Chicago, Illinois, United States
  • Mark Rosenblatt
    Illinois Eye and Ear Infirmary, UIC, Chicago, Illinois, United States
  • Ali R Djalilian
    Illinois Eye and Ear Infirmary, UIC, Chicago, Illinois, United States
  • Footnotes
    Commercial Relationships   Ghasem Yazdanpanah, None; Meisam Omidi, None; Ritu Shah, None; Sayena Jabbehdari, None; Khandaker Anwar, None; Xiang Shen, None; Behnam Rabiee, None; Ilham Putra, None; Mark Rosenblatt, None; Ali Djalilian, None
  • Footnotes
    Support  This work was supported by R01 EY024349 (ARD) and Core Grant for Vision Research EY01792 (MIR) from NEI/NIH; unrestricted grant to the department from Research to Prevent Blindness.
Investigative Ophthalmology & Visual Science June 2020, Vol.61, 1198. doi:
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      Ghasem Yazdanpanah, Meisam Omidi, Ritu Shah, Sayena Jabbehdari, Khandaker N Anwar, Xiang Shen, Behnam Rabiee, Ilham Putra, Mark Rosenblatt, Ali R Djalilian; Production of Bio-active Nanoparticle from Human Corneal Extracellular Matrix. Invest. Ophthalmol. Vis. Sci. 2020;61(7):1198.

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

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Abstract

Purpose : Extracellular matrix (ECM) is composed of structural proteins and glycans as well as bio-active molecules such as growth factors, peptides and nucleic acids. Recent studies have shown the regenerative potential of processed decellularized ECMs derived from different organs. Here, we propose a novel protocol to produce nanoparticles with preserved bioactivity from human corneas for future corneal regenerative and tissue engineering applications.

Methods : Human cadaver corneas were kindly provided by Eversight Eye Bank. After washing, the corneas were cut into small pieces and decellularized by several cycles of freeze-thawing and nuclease treatment. The decellularized cornea pieces were then lyophilized and cryo-milled into microparticles (<230 µm particle size). The ECM microparticles were turned into nanoparticles by stirring in 5% acetic acid (5 mg/ml) followed by probe sonication. Various durations of acid treatment and sonication were applied to optimize the protocol. The particle sizes were measured using Dynamic Light Scattering. The proliferation of Human Corneal Epithelial Cells (HCECs) treated with 0.5 mg/ml human cornea ECM (HC-ECM) nanoparticles compared to control was assessed using a Cell Counting Kit to evaluate the bioactivity of produced samples.

Results : The average size of HC-ECM nanoparticles progressively decreased with increased time of acetic acid treatment (1645.3±373.5 nm, 843.5±301.4 nm and 390.9±331.4 nm after 24, 96 and 168 h, respectively). However, the mean polydispersity index (PDI) increased which indicates the reduction in uniformity of produced nanoparticles (0.85±0.16, 0.92±0.12 and 1±0.5 after 24, 96 and 168 h, respectively). Probe sonication not only decreased the size of particles but also improved the PDI in time (size, 493.7± 15.8nm, 485.8± 86.8nm, 314.3± 44.6nm and PDI, 0.58± 0.04, 0.4± 0.06, 0.48± 0.01 after 10, 20 and 30 minutes, respectively). We found that 24 hours stirring in 5% acetic acid followed by probe sonication for 20 minutes is the optimized protocol to produce ECM nanoparticles with highest uniformity. The proliferation of HCECs was significantly improved after treatment with HC-ECM nanoparticles compared to controls (P<0.001).

Conclusions : The proposed protocol for producing bioactive HC-ECM nanoparticles provides the potential opportunity of recycling unsuitable human corneas, for transplantation, into the therapeutic applications.

This is a 2020 ARVO Annual Meeting abstract.

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