Collagen crosslinking-induced corneal morphological changes: a three-dimensional light sheet Microscopy-based evaluation.

Publisher: Nature Publishing Group Country of Publication: England NLM ID: 101563288 Publication Model: Electronic Cited Medium: Internet ISSN: 2045-2322 (Electronic) Linking ISSN: 20452322 NLM ISO Abbreviation: Sci Rep Subsets: MEDLINE

Original Publication: London : Nature Publishing Group, copyright 2011-

Collagen*/metabolism ; Imaging, Three-Dimensional*/methods ; Cornea*/drug effects ; Cornea*/diagnostic imaging ; Cornea*/metabolism ; Microscopy, Fluorescence*/methods; Animals ; Swine ; Cross-Linking Reagents/pharmacology ; Keratoconus/metabolism ; Keratoconus/drug therapy ; Keratoconus/pathology ; Glutaral

Stiffness-related eye diseases such as keratoconus require comprehensive visualization of the complex morphological matrix changes. The aim of this study was to use three-dimensional (3D) light sheet fluorescence microscopy (LSFM) to analyze unlabeled corneal tissue samples, qualitatively visualizing changes in corneal stiffness. Isolated porcine corneal tissue samples were treated with either NaCl or 0.1% glutaraldehyde (GTA) prior to clearing with benzyl alcohol/benzyl benzoate (BABB) and subsequently scanned with LSFM. After analysis of the LSFM data sets, the samples were embedded in paraffin to validate the results by conventional planar microscopy. In the unlabeled corneal tissue samples the 2D/3D morphology of the entire tissue volume was identified by specific autofluorescence signals. An enhancement of collagen crosslinking was induced by applying GTA to the corneal tissue. Subsequent LSFM scans showed specific morphological changes due to altered autofluorescence signals of the corneal stroma, which were confirmed by conventional histology. Therefore, LSFM analysis of corneal tissue samples allowed label-free 3D autofluorescence assessment of the corneal morphology in its anatomical context. It provides the technical basis for the examination of the pathologically altered cornea and facilitates ophthalmologic examinations of corneal diseases based on the altered tissue stiffness.
Competing Interests: Declarations Competing interests The authors declare no competing interests.
(© 2024. The Author(s).)

JAMA Ophthalmol. 2016 Feb;134(2):167-73. (PMID: 26633035)
Elife. 2020 Feb 19;9:. (PMID: 32073398)
Andrology. 2022 Nov;10(8):1660-1672. (PMID: 36082398)
Ocul Surf. 2023 Oct;30:150-159. (PMID: 37683969)
Sci Rep. 2020 Oct 26;10(1):18246. (PMID: 33106532)
J Histochem Cytochem. 2014 Oct;62(10):751-8. (PMID: 25023614)
Exp Eye Res. 2016 May;146:137-144. (PMID: 26677758)
An Bras Dermatol. 2018 Jun;93(3):415-418. (PMID: 29924246)
J Biophotonics. 2018 Jul;11(7):e201700324. (PMID: 29575612)
Acta Crystallogr Sect F Struct Biol Cryst Commun. 2010 Mar 1;66(Pt 3):225-8. (PMID: 20208148)
Exp Eye Res. 2019 Mar;180:137-145. (PMID: 30578790)
Exp Eye Res. 1998 Jan;66(1):97-103. (PMID: 9533835)
Ophthalmologica. 2014;232(1):10-27. (PMID: 24751584)
Ocul Surf. 2020 Jul;18(3):526-532. (PMID: 32417103)
Clin Radiol. 2018 Jun;73(6):595.e1-595.e7. (PMID: 29510833)
Arch Soc Esp Oftalmol (Engl Ed). 2023 Apr;98(4):206-212. (PMID: 36871853)
Nat Protoc. 2012 Nov;7(11):1983-95. (PMID: 23060243)
Asian J Androl. 2014 Mar-Apr;16(2):305-8. (PMID: 24435054)
FEBS Lett. 1986 Mar 3;197(1-2):55-8. (PMID: 3512309)
J Invest Dermatol. 1968 May;50(5):419-23. (PMID: 4172462)
Klin Monbl Augenheilkd. 2022 Dec;239(12):1427-1432. (PMID: 35977709)
Annu Rev Neurosci. 1992;15:1-29. (PMID: 1575438)
Sci Rep. 2020 Oct 29;10(1):18594. (PMID: 33122764)
Taiwan J Ophthalmol. 2024 Feb 02;14(1):44-49. (PMID: 38654993)
Eye (Lond). 2017 Oct;31(10):1496-1502. (PMID: 28574496)
BMC Ophthalmol. 2018 Apr 13;18(1):94. (PMID: 29653563)
Klin Monbl Augenheilkd. 2020 Dec;237(12):1442-1454. (PMID: 33231276)
Mater Sci Eng C Mater Biol Appl. 2019 Mar;96:941-954. (PMID: 30606606)
Skin Res Technol. 2018 May;24(2):294-303. (PMID: 29377352)
Int J Ophthalmol. 2024 May 18;17(5):951-962. (PMID: 38766341)
Histochem Cell Biol. 2016 Dec;146(6):781-806. (PMID: 27704211)
J Biomed Opt. 2018 Mar;23(3):1-8. (PMID: 29500874)
Cell. 2014 Nov 6;159(4):911-24. (PMID: 25417165)
Cornea. 2016 Nov;35 Suppl 1:S31-S37. (PMID: 27631351)
J Biomed Opt. 2017 Jul 1;22(7):76011. (PMID: 28717817)
Sci Rep. 2019 Jul 15;9(1):10241. (PMID: 31308406)
Histochem J. 1979 Jul;11(4):447-55. (PMID: 91593)
Sci Rep. 2020 Oct 19;10(1):17619. (PMID: 33077794)

Keywords: 3D virtual histology; Autofluorescence; Corneal stroma; Extracellular matrix stiffness; Light sheet microscopy; Tomographic microscopy

9007-34-5 (Collagen)
0 (Cross-Linking Reagents)
T3C89M417N (Glutaral)

Date Created: 20241116 Date Completed: 20241116 Latest Revision: 20241122

20250114

PMC11569206

10.1038/s41598-024-78516-x

39550403