Y47 - Biocompatible Microgels from Decellularized Porcine Ocular Tissue for Retinal Cell Support

Saturday, October 11, 2025

10:00 AM - 11:00 AM PDT

Location: Exhibit Hall F, G & H

Presenting Author(s)

AJ

Anna K. Jenks

Student
North Carolina State University
Camden, South Carolina, United States

Co-Author(s)

ZW

Ziyu Wang

Post-doc
NCSU/UNC Joint Department of Biomedical Engineering
Raleigh, North Carolina, United States

Primary Investigator(s)

Donald Freytes

Associate Professor
NCSU/UNC Lampe Joint Department of Biomedical Engineering
Raleigh, North Carolina, United States

Introduction: : Age-related Macular Degeneration (AMD) is a leading cause of vision loss and affects an estimated 19.8 million Americans over age 40. The progressive deterioration of the macula is a characteristic of AMD and occurs in two forms, wet and dry, with the latter being the more common. Despite the prevalence and severity of dry AMD, current treatments remain limited and do not address the progressive degeneration of retinal structure and function. Introducing conductive properties to the retina may help restore visual function by supporting electrochemical signaling and aiding the performance of damaged photoreceptors, Retinal Pigmented Epithelial (RPE) cells, and Retinal Ganglion Cells (RGC). Porcine eyes serve as a practical alternative tissue source for ocular repair due to their anatomical similarity to human eyes and have been used in this investigation. This project aims to evaluate the effectiveness of processing decellularized porcine ocular tissues into bioactive microgels that support cellular viability. These biocompatible microgels will facilitate the delivery of therapeutic agents to promote tissue repair within the retina and optic nerve.

Materials and

Methods: : Porcine eyes are collected and dissected to isolate the cornea and optic nerve tissue under sterile conditions. This tissue is then decellularized with 4% SDC, DNase, and 0.1% Peracetic acid to preserve bioactive ECM components. The decellularization process is validated using PicoGreen assays to quantify the removal of dsDNA. After decellularization, the tissue is frozen, ground, and lyophilized to produce a fine powder of decellularized extracellular matrix (dECM). Hydrogels are chemically crosslinked using GelMA, PEGDA, PEDOT:PSS, and Irgacure. The hydrogels are cured using UV light, and a photoinitiator is incorporated into the gel. The physical properties of the hydrogel, including stiffness, elasticity, conductivity, and degree of crosslinking, are measured. Both RPE and RGC cells are isolated and cultured to test the cell viability of the hydrogels. Verification of RPE cell isolation is performed using immunostaining for RPE-specific markers.

Results, Conclusions, and Discussions:: The decellularization protocol effectively removed significant amounts of DNA from native ocular tissue. Quantification using the PicoGreen assay showed that 90.84% of DNA was removed from the decellularized cornea tissue, and 87.86% was removed from the decellularized optic nerve tissue. This indicates a successful decellularization process and suggests a potential reduction in immunogenicity for therapeutic applications. RGC cells were successfully isolated, whereas RPE cells exhibited slower proliferation than expected. Hydrogels formed from decellularized tissue were successfully crosslinked and cured to produce microgels with tunable and measurable mechanical properties. Optimal concentrations were identified and used to produce hydrogels with sufficient mechanical strength and softness for ocular applications. Future work will focus on measuring the mechanical properties of the hydrogels and incorporating conductive materials, such as carbon nanotubes, to aid in the detection of electrochemical signals within the eye. RPE cell culturing showed limited proliferation and minimal RPE cell markers during immunostaining. This suggests errors in the isolation protocol and cell digestion. Future improved protocols would focus on effective digestion and the optimization of cell culture media. This study demonstrates that porcine ocular tissue can be decellularized and reconstituted into biocompatible microgels. These findings provide a foundation for developing bioactive scaffolds that support photoreceptors and retinal cells in diseases such as AMD.

Acknowledgements and/or References (Optional): :