Logo image
Development of High-Resolution Three-Dimensional-Printed Extracellular Matrix Scaffolds and Their Compatibility with Pluripotent Stem Cells and Early Retinal Cells
Journal article   Open access   Peer reviewed

Development of High-Resolution Three-Dimensional-Printed Extracellular Matrix Scaffolds and Their Compatibility with Pluripotent Stem Cells and Early Retinal Cells

Arwin Shrestha, Brittany N Allen, Luke A Wiley, Budd A Tucker and Kristan S Worthington
Journal of ocular pharmacology and therapeutics, Vol.36(1), pp.42-55
01/2020
DOI: 10.1089/jop.2018.0146
PMCID: PMC7476392
PMID: 31414943
url
https://doi.org/10.1089/jop.2018.0146View
Published (Version of record) Open Access

Abstract

Widely used approaches for retinal disease modeling and therapeutic testing can be augmented by using tissue-engineered scaffolds with a precise 3-dimensional structure. However, the materials currently used for these scaffolds are poorly matched to the biochemical and mechanical properties of the retina. Here, we create biopolymer-based scaffolds with a structure that is amenable to retinal tissue engineering and modeling. Optimal two-photon polymerization (TPP) settings, including laser power and scanning speed, are identified for 4 methacrylated biopolymer formulations: collagen, gelatin, hyaluronic acid (HA), and a 50/50 mixture of gelatin/HA, each with methylene blue as a photoinitiator. For select formulations, fabrication accuracy and swelling are determined and biocompatibility is evaluated by using human induced pluripotent stem cells and rat postnatal retinal cells. TPP is feasible for each biopolymer formulation, but it is the most reliable for mixtures containing gelatin and the least reliable for HA alone. The mean size of microscaffold pores is within several microns of the intended value but the overall structure size is several times greater than the modeled volume. The addition of HA to gelatin scaffolds increases cell viability and promotes neuronal phenotype, including Tuj-1 expression and characteristic morphology. We successfully determined a useful range of TPP settings for 4 methacrylated biopolymer formulations. When crosslinked, these extracellular matrix-derived molecules support the growth and attachment of retinal cells. We anticipate that when combined with existing patient-specific approaches, this technique will enable more efficient and accurate retinal disease modeling and therapeutic testing than current techniques allow.
Tissue Engineering Retina - metabolism Cell Survival Humans Cells, Cultured Hyaluronic Acid - chemistry Extracellular Matrix - metabolism Polymerization Rats Photons Rats, Sprague-Dawley Extracellular Matrix - chemistry Animals Retina - cytology Hyaluronic Acid - metabolism Gelatin - metabolism Induced Pluripotent Stem Cells - cytology Printing, Three-Dimensional Gelatin - chemistry Induced Pluripotent Stem Cells - metabolism

Details

Metrics

Logo image