ORCID
Alison E. Patteson: 0000-0002-4004-1734
Document Type
Article
Date
Winter 1-29-2021
Keywords
In vitro, Bioengineering, Tissue stiffening, ECM mechanics, POAG
Language
English
Funder(s)
American Heart Association grant, National Institutes of Health grants, SUNY Upstate Medical University Department of Ophthalmology and Visual Sciences from Research to Prevent Blindness (RPB) and from Lions Region 20-Y1, and RPB Career Development Award
Funding ID
16SDG31280010, EB023052, HL140618, EY031710, EY030617 and EY028608
Acknowledgements
We thank Iris Navarro at the Duke Eye Center - BioSight Program for guidance in setting up initial HTM isolation and characterization procedures. We also thank the team at Specialty Surgery Center of Central New York for assistance with corneal rim specimens. We thank Drs. Audrey M. Bernstein, Mariano S. Viapiano, and Jason A. Horton for imaging support. We also thank the teams at the Syracuse University - Syracuse Biomaterials Institute and BioInspired Institute for access to shared facilities and technical advice. We thank Benjamin Zink and Debra Driscoll at the SUNY College of Environmental Science and Forestry - Analytical and Technical Services for assistance with SEM imaging. Author contributions: H.L., A.N.S., H.R., A.E.P., W.D.S., P.S.G., and S.H designed all experiments, collected, analyzed, and interpreted the data. T.B. and A.K. assisted with HTM cell isolation, ELP expression/purification, PDMS-mold design/3D printing, and imaging. R.W.W., N.A., and W.D.S. provided study materials. H.L. and S.H. wrote the manuscript. All authors collected data and commented on and approved the final manuscript. P.S.G. and S.H. conceived and supervised the research. Competing interests: The authors declare no conflict of interest. Data and materials availability: All data needed to evaluate the conclusions in the paper are present in the paper and/or the Supplementary Materials. Additional data related to this paper may be requested from the authors.
Official Citation
Li H, Bagué T, Kirschner A, Strat AN, Roberts H, Weisenthal RW, Patteson AE, Annabi N, Stamer WD, Ganapathy PS, Herberg S. A tissue-engineered human trabecular meshwork hydrogel for advanced glaucoma disease modeling. Exp Eye Res. 2021 Apr;205:108472. doi: 10.1016/j.exer.2021.108472. Epub 2021 Jan 29. PMID: 33516765.
Disciplines
Physics
Description/Abstract
Abnormal human trabecular meshwork (HTM) cell function and extracellular matrix(ECM) remodeling contribute to HTM stiffening in primary open-angle glaucoma (POAG). Most current cellular HTM model systems do not sufficiently replicate the complex native three dimensional (3D) cell-ECM interface, limiting their use for investigating POAG pathology. Tissue-engineered hydrogels are ideally positioned to overcome shortcomings of current models. Here, we report a novel biomimetic HTM hydrogel and test its utility as a POAG disease model. HTM hydrogels were engineered by mixing normal donor-derived HTM cells with collagen type I, elastin-like polypeptide and hyaluronic acid, each containing photoactive functional groups, followed by UV crosslinking. Glaucomatous conditions were induced with dexamethasone (DEX), and effects of the Rho-associated kinase (ROCK) inhibitor Y27632 on cytoskeletal organization and tissue-level function, contingent on HTM cell-ECM interactions, were assessed. DEX exposure increased HTM hydrogel contractility, f-actin and alpha smooth muscle actin abundance and rearrangement, ECM remodeling, and fibronectin deposition - all contributing to HTM hydrogel condensation and stiffening consistent with glaucomatous HTM tissue behavior. Y27632 treatmentproduced precisely the opposite effects and attenuated the DEX-induced pathologic changes, resulting in HTM hydrogel relaxation and softening. For model validation, confirmed glaucomatous HTM (GTM) cells were encapsulated; GTM hydrogels showed increased contractility, fibronectin deposition, and stiffening vs. normal HTM hydrogels despite reduced GTM cell proliferation. We have developed a biomimetic HTM hydrogel model for detailed investigation of 3D cell-ECM interactions under normal and simulated glaucomatous conditions. Its bidirectional responsiveness to pharmacological challenge and rescue suggests promising potential to serve as screening platform for new POAG treatments with focus on HTM biomechanics.
Recommended Citation
Li H, Bagué T, Kirschner A, Strat AN, Roberts H, Weisenthal RW, Patteson AE, Annabi N, Stamer WD, Ganapathy PS, Herberg S. A tissue-engineered human trabecular meshwork hydrogel for advanced glaucoma disease modeling. Exp Eye Res. 2021 Apr;205:108472. doi: 10.1016/j.exer.2021.108472. Epub 2021 Jan 29. PMID: 33516765.
Creative Commons License
This work is licensed under a Creative Commons Attribution 4.0 International License.
