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A method for quantifying limbal stem cell niches using OCT imaging
  1. Michel Haagdorens1,2,
  2. Joséphine Behaegel1,3,
  3. Jos Rozema1,2,
  4. Veerle Van Gerwen1,
  5. Sofie Michiels2,
  6. Sorcha Ní Dhubhghaill1,2,
  7. Marie-José Tassignon1,2,
  8. Nadia Zakaria1,2,4
  1. 1 Faculty of Medicine and Health Sciences, Department of Ophthalmology, Visual Optics and Visual Rehabilitation, Antwerp University, Antwerp, Wilrijk, Belgium
  2. 2 Department of Ophthalmology, Antwerp University Hospital, Edegem, Belgium
  3. 3 Department of Ophthalmology, University Hospital Brussels, Brussels, Belgium
  4. 4 Center for Cell Therapy and Regenerative Medicine, Antwerp University Hospital, Edegem, Belgium
  1. Correspondence to Dr Nadia Zakaria, Department of Ophthalmology, Visual Optics and Visual Rehabilitation, University of Antwerp, Universiteitsplein 1, Antwerpen Campus Drie Eiken –Gebouw T T4—Oogheelkunde, Antwerp 2610, Belgium; nadia.zakaria{at}uza.be

Abstract

Aims To evaluate the efficacy of Fourier domain-optical coherence tomography (FD-OCT) in imaging and quantifying the limbal palisades of Vogt and to correlate these images with histological findings.

Methods The superior and inferior limbal region of both eyes of 50 healthy volunteers were imaged by FD-OCT. Images were processed and analysed using Matlab software. In vitro immunofluorescent staining of a cadaveric donor limbus was analysed to correlate the presence of stem cells in the visualised structures.

Results FD-OCT could successfully visualise limbal crypts and the palisades of Vogt in the limbus region. Fluorescent labelling confirmed the presence of stem cells in these structures. The mean palisade ridge width (ΔPR) and the mean interpalisade epithelial rete peg width (ΔERP) were both of the order of 72 μm, leading to a palisade density (PD) of about 7.4 palisades/mm. A significant difference in ΔPR, ΔERP and PD was seen between the inferior and superior sides of the right eye and the superior sides of the left and right eye(p<0.05.). A significant influence of iris colour on parameters ΔPR, ΔERP and PD was found, and of age on PD and ΔERP (p<0.05).

Conclusions In vivo OCT imaging is a safe and effective modality to image the limbus and can be used to visualise the palisades of Vogt. Image processing using Matlab software enabled quantification and density calculation of imaged limbal palisades of Vogt. This technique may enhance targeted limbal biopsies for transplantation.

  • Cornea
  • Imaging
  • Physiology
  • Stem Cells

This is an Open Access article distributed in accordance with the Creative Commons Attribution Non Commercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited and the use is non-commercial. See: http://creativecommons.org/licenses/by-nc/4.0/

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Footnotes

  • MH and JB contributed equally.

  • Contributors MH, JR, SND, M-JT and NZ contributed to the planning of this study. MH, JB, JR, VVG, SM and NZ contributed to the conduction of this study, whereas MH, JB, JR, SND and NZ significantly contributed to the reporting of the work.

  • Funding This research was funded by, Horizon2020 ARREST BLINDNESS, ‘Agentschap voor Innovatie door Wetenschap en Technologie—Toegepast Biomedisch onderzoek’ (IWT-TBM), and Funds for Research in Ophthalmology Belgium. Michel Haagdorens is a PhD student who holds a Research Foundation—Flanders scholarship.

  • Competing interests None declared.

  • Patient consent Obtained.

  • Ethics approval Ethisch Comité UZA.

  • Provenance and peer review Not commissioned; externally peer reviewed.

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