Decellularizing the porcine optic nerve head: Toward a model to study the mechanobiology of glaucoma

Jr Jiun Liou, Michelle D. Drewry, Ashlinn Sweeney, Bryan N. Brown, Jonathan P. Vande Geest

Research output: Contribution to journalArticlepeer-review

3 Scopus citations

Abstract

Purpose: Studying the extracellular matrix (ECM) remodeling of the lamina cribrosa in vivo can be extremely challenging and costly. There exist very few options for studying optic nerve head (ONH) mechanobiology in vitro that are able to reproduce the complex anatomic and biomechanical environment of the ONH. Herein, we have developed a decellularization procedure that will enable more anatomically relevant and cost-efficient future studies of ECM remodeling of the ONH. Methods: Porcine posterior poles were decellularized using a detergent and enzyme-based decellularization protocol. DNA quantification and histology were used to inves-tigate the effectiveness of the protocol. We subsequently investigated the ability of a polyethylene glycol (PEG)-based hydrogel to restore the ONH’s ability to hold pressure following decellularization. Anterior-posterior displacement of the decellularized and PEG treated ONH in a pressure bioreactor was used to evaluate the biomechanical response of the ONH. Results: DNA quantification and histology confirmed decellularization using Triton X-100 at low concentration for 48 hours successfully reduced the cellular content of the tissue by 94.9% compared with native tissue while preserving the ECM microstructure and basal lamina of the matrix. Infiltrating the decellularized tissues with PEG 6000 and PEG 10,000 hydrogel restored their ability to hold pressure, producing displacements similar to those measured for the non-decellularized control samples. Conclusions: Our decellularized ONH model is capable of producing scaffolds that are cell-free and maintain the native ECM microstructure. Translational Relevance: This model represents a platform to study the mechanobiol-ogy in the ONH and potentially for glaucoma drug testing.

Original languageEnglish (US)
Article number17
JournalTranslational Vision Science and Technology
Volume9
Issue number8
DOIs
StatePublished - 2020
Externally publishedYes

Keywords

  • Decellularization
  • Glaucoma
  • Lamina cribrosa
  • Mechanobiology
  • Porcine

ASJC Scopus subject areas

  • Biomedical Engineering
  • Ophthalmology

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