Freeze-thaw decellularization of the trabecular meshwork in an ex vivo eye perfusion model

PeerJ
Yalong DangNils Loewen

Abstract

The trabecular meshwork (TM) is the primary substrate of outflow resistance in glaucomatous eyes. Repopulating diseased TM with fresh, functional TM cells might be a viable therapeutic approach. Decellularized TM scaffolds have previously been produced by ablating cells with suicide gene therapy or saponin, which risks incomplete cell removal or dissolution of the extracellular matrix, respectively. We hypothesized that improved trabecular meshwork cell ablation would result from freeze-thaw cycles compared to chemical treatment. We obtained 24 porcine eyes from a local abattoir, dissected and mounted them in an anterior segment perfusion within two hours of sacrifice. Intraocular pressure (IOP) was recorded continuously by a pressure transducer system. After 72 h of IOP stabilization, eight eyes were assigned to freeze-thaw (F) ablation (-80 °C × 2), to 0.02% saponin (S) treatment, or the control group (C), respectively. The TM was transduced with an eGFP expressing feline immunodeficiency viral (FIV) vector and tracked via fluorescent microscopy to confirm ablation. Following treatment, the eyes were perfused with standard tissue culture media for 180 h. TM histology was assessed by hematoxylin and eosin staining. TM viabilit...Continue Reading

References

Feb 1, 1990·The British Journal of Ophthalmology·M T Benson, M E Nelson
Dec 19, 2001·Human Gene Therapy·Nils LoewenEric M Poeschla
Mar 1, 1953·Biochimica Et Biophysica Acta·J E LOVELOCK
May 17, 2005·BJU International·J G Baust, A A Gage
Jan 24, 2007·Biochimica Et Biophysica Acta·Willem F WolkersJohn C Bischof
Jun 16, 2010·Archives of Ophthalmology·Alberto IzzottiCristina Cartiglia
Aug 7, 2010·Journal of Vascular and Interventional Radiology : JVIR·Joseph P Erinjeri, Timothy W I Clark
Dec 3, 2010·Journal of Cell Science·Christian FrantzValerie M Weaver
Feb 3, 2011·Graefe's Archive for Clinical and Experimental Ophthalmology = Albrecht Von Graefes Archiv Für Klinische Und Experimentelle Ophthalmologie·Irene SanchezIvan Fernandez-Bueno
Nov 16, 2012·Nature·Martien A M GroenenLawrence B Schook
Dec 20, 2012·Investigative Ophthalmology & Visual Science·Jose M GonzalezJames C H Tan
Jan 24, 2013·Investigative Ophthalmology & Visual Science·Yiqin DuJoel S Schuman
Jul 13, 2013·Investigative Ophthalmology & Visual Science·Julius T OattsNils Loewen
Dec 10, 2013·Nucleic Acids Research·Paul FlicekStephen M J Searle
Jan 23, 2014·Investigative Ophthalmology & Visual Science·Ze ZhangNils Loewen
Feb 25, 2014·Nature Reviews. Cancer·Katrina F Chu, Damian E Dupuy
Sep 13, 2014·Journal of Cellular Physiology·Sergio Claudio SaccàAlberto Izzotti
Dec 17, 2014·The American Journal of Pathology·Padmanabhan P PattabiramanPonugoti V Rao
Jun 13, 2015·Investigative Ophthalmology & Visual Science·Joseph C PetersGulab S Zode
Sep 5, 2015·Graefe's Archive for Clinical and Experimental Ophthalmology = Albrecht Von Graefes Archiv Für Klinische Und Experimentelle Ophthalmologie·Isabel GorslerChristian Meltendorf
Nov 29, 2015·Biotechnology and Bioengineering·Karen Y TorrejonYubing Xie
Jan 7, 2016·The British Journal of Ophthalmology·Kevin KaplowitzNils Loewen
Mar 24, 2016·Investigative Ophthalmology & Visual Science·Ralitsa T LoewenNils Loewen
May 2, 2016·Experimental Eye Research·Yalong DangNils Loewen
May 18, 2016·Journal of Ocular Pharmacology and Therapeutics : the Official Journal of the Association for Ocular Pharmacology and Therapeutics·Hongmin YunYiqin Du
May 27, 2016·Investigative Ophthalmology & Visual Science·Ralitsa T LoewenNils Loewen
Jun 9, 2016·Proceedings of the National Academy of Sciences of the United States of America·Wei ZhuMarkus H Kuehn

Citations

Apr 5, 2019·Graefe's Archive for Clinical and Experimental Ophthalmology = Albrecht Von Graefes Archiv Für Klinische Und Experimentelle Ophthalmologie·Ying HongNils Loewen
Nov 21, 2018·Graefe's Archive for Clinical and Experimental Ophthalmology = Albrecht Von Graefes Archiv Für Klinische Und Experimentelle Ophthalmologie·Yalong DangNils Loewen
Aug 28, 2020·APL Bioengineering·Wenzhen ZhuDong-An Wang
Aug 29, 2020·Translational Vision Science & Technology·Jr-Jiun LiouJonathan P Vande Geest
May 31, 2018·Investigative Ophthalmology & Visual Science·Susannah WaxmanNils Loewen
May 4, 2018·Graefe's Archive for Clinical and Experimental Ophthalmology = Albrecht Von Graefes Archiv Für Klinische Und Experimentelle Ophthalmologie·Yalong DangNils Loewen

Related Concepts

Extracellular Matrix
Eye
Gene Therapy
Genetic Vectors
Hematoxylin
Physiologic Intraocular Pressure
Microscopy, Fluorescence
Propidium Iodide
Saponins
Family suidae

Related Feeds

Cell Atlas of the Human Eye

Constructing a cell atlas of the human eye will require transcriptomic and histologic analysis over the lifespan. This understanding will aid in the study of development and disease. Find the latest research pertaining to the Cell Atlas of the Human Eye here.

Related Papers

Cellular Physiology and Biochemistry : International Journal of Experimental Cellular Physiology, Biochemistry, and Pharmacology
Huan ZouJian Ye
© 2021 Meta ULC. All rights reserved