September 2016
Volume 57, Issue 12
Open Access
ARVO Annual Meeting Abstract  |   September 2016
Design and validation of a compact ex vivo whole eye perfusion system
Author Affiliations & Notes
  • Enhua Zhou
    Novartis Institutes for BioMedical Research, Cambridge, Massachusetts, United States
  • Michael Paolucci
    Novartis Institutes for BioMedical Research, Cambridge, Massachusetts, United States
  • sam shrestha
    Novartis Institutes for BioMedical Research, Cambridge, Massachusetts, United States
  • Christopher William Wilson
    Novartis Institutes for BioMedical Research, Cambridge, Massachusetts, United States
  • Ernie Hixon
    Novartis Institutes for BioMedical Research, Cambridge, Massachusetts, United States
  • Andrew Brady
    Novartis Institutes for BioMedical Research, Cambridge, Massachusetts, United States
  • Aaron Bickel
    Novartis Institutes for BioMedical Research, Cambridge, Massachusetts, United States
  • Ted Manley
    Novartis Institutes for BioMedical Research, Cambridge, Massachusetts, United States
  • Ganesh Prasanna
    Novartis Institutes for BioMedical Research, Cambridge, Massachusetts, United States
  • Dennis S Rice
    Novartis Institutes for BioMedical Research, Cambridge, Massachusetts, United States
  • Amy Chen
    Novartis Institutes for BioMedical Research, Cambridge, Massachusetts, United States
  • Footnotes
    Commercial Relationships   Enhua Zhou, Novartis Institutes of BioMedical Research (E); Michael Paolucci, Novartis Institutes for BioMedical Research (E); sam shrestha, Novartis Institutes for BioMedical Research (E); Christopher Wilson, Novartis Institutes for BioMedical Research (E); Ernie Hixon, Novartis Institutes for BioMedical Research (E); Andrew Brady, Novartis Institutes for BioMedical Research (E); Aaron Bickel, Novartis Institutes for BioMedical Research (E); Ted Manley, Novartis Institutes for BioMedical Research (E); Ganesh Prasanna, Novartis Institutes for BioMedical Research (E); Dennis Rice, Novartis Institutes for BioMedical Research (E); Amy Chen, Novartis Institutes of BioMedical Research (E)
  • Footnotes
    Support  None
Investigative Ophthalmology & Visual Science September 2016, Vol.57, No Pagination Specified. doi:
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      Enhua Zhou, Michael Paolucci, sam shrestha, Christopher William Wilson, Ernie Hixon, Andrew Brady, Aaron Bickel, Ted Manley, Ganesh Prasanna, Dennis S Rice, Amy Chen; Design and validation of a compact ex vivo whole eye perfusion system. Invest. Ophthalmol. Vis. Sci. 201657(12):.

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

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Abstract

Purpose : A clinically validated, manageable risk factor for glaucoma progression is elevated Intraocular pressure (IOP). IOP is maintained by the balance between aqueous humor production and outflow resistance. To investigate the regulation of outflow resistance, ex vivo perfusion of animal eyes provides a valuable tool. However, typical whole eye perfusion systems reported in glaucoma research have limited throughput (e.g. 1-2 pairs a time). Here we report the design of a compact, scalable perfusion system with improved throughput. This is achieved by incorporating flow sensors into a gravity-driven perfusion system.

Methods : IOP was controlled by the height of the reservoir, which was adjusted by a motorized stage. Flow rate was measured by flow sensors (Sensirion, Switzerland). Custom software was developed in Matlab (Mathworks, Natick, MA) to control reservoir height and to record flow rate. We validated the system by measuring response of eyes to Y-39983 (a Rho kinase inhibitor), latrunculin B (actin disrupting agent), prostaglandin E1 (PGE1), endothelin-1 (ET-1; potent vasoactive peptide), ambrisentan (inhibitor for type A endothelin receptor), and S-nitroso-N-acetylpenicillamine (SNAP, a nitric oxide [NO] donor). All compounds were evaluated in bovine eyes and a few selected ones were also tested in rabbit and porcine eyes.

Results : The instrument design enabled simultaneous measurements of 10 eyes with a small footprint of 4 square feet. At fixed IOP of 6 mmHg, Y-39983, Latrunculin B, or PGE1 increased outflow rate by up to 50-100%, compared to vehicle control, in a time- and dose-dependent manner. On the contrary, ET-1 decreased outflow rate by up to 50-70% with an EC50 around 7nM. ET-1’s effect was inhibited by ambrisentan as expected. Interestingly, it was also inhibited by pretreatment of SNAP (by up to 84% in bovine eyes) or Y-39983 (by 21% in porcine eyes and 100% in rabbit eyes).

Conclusions : We have developed a compact system that can reliably measure outflow resistance in ex vivo cultured eyes. Results from Y-39983, latrunculin B, and PGE1 recapitulated in vivo animal studies reported in the literature. This platform offers an opportunity in enabling compound prescreen for in-vivo efficacy studies for novel glaucoma therapies.

Acknowledgement: We thank Tom Vollmer for technical assistance.

This is an abstract that was submitted for the 2016 ARVO Annual Meeting, held in Seattle, Wash., May 1-5, 2016.

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