June 2023
Volume 64, Issue 8
Open Access
ARVO Annual Meeting Abstract  |   June 2023
Corneal haze clearance mechanism and visual function recovery following stromal keratocyte injection
Author Affiliations & Notes
  • Andri K Riau
    Tissue Engineering & Cell Therapy Group, Singapore Eye Research Institute, Singapore, Singapore
    EYE-ACP, Duke-NUS Medical School, Singapore, Singapore
  • Evelina Han
    Tissue Engineering & Cell Therapy Group, Singapore Eye Research Institute, Singapore, Singapore
  • Gary Yam
    Department of Ophthalmology, University of Pittsburgh, Pittsburgh, Pennsylvania, United States
  • Craig Boote
    School of Optometry & Vision Sciences, Cardiff University, Cardiff, United Kingdom
  • Hon Shing Ong
    Tissue Engineering & Cell Therapy Group, Singapore Eye Research Institute, Singapore, Singapore
    EYE-ACP, Duke-NUS Medical School, Singapore, Singapore
  • Jodhbir S Mehta
    Tissue Engineering & Cell Therapy Group, Singapore Eye Research Institute, Singapore, Singapore
    EYE-ACP, Duke-NUS Medical School, Singapore, Singapore
  • Footnotes
    Commercial Relationships   Andri Riau None; Evelina Han None; Gary Yam None; Craig Boote None; Hon Shing Ong None; Jodhbir Mehta None
  • Footnotes
    Support  None
Investigative Ophthalmology & Visual Science June 2023, Vol.64, 3810. doi:
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      Andri K Riau, Evelina Han, Gary Yam, Craig Boote, Hon Shing Ong, Jodhbir S Mehta; Corneal haze clearance mechanism and visual function recovery following stromal keratocyte injection. Invest. Ophthalmol. Vis. Sci. 2023;64(8):3810.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose : Cornea, the transparent and collagenous structure that allows vision, is susceptible to trauma and infections, resulting in fibrosis and, in many cases, blindness. We previously showed that the injection of cultured human stromal keratocytes (CSKs) resolved acute corneal haze in a rat model of irregular phototherapeutic keratectomy (irrPTK). Here, we investigated the mechanism of haze clearance and the recovery of visual functions in those rats.

Methods : The haze manifested 1 week following irrPTK on the rat cornea. A total of 40,000 CSKs were then injected into the haze region. The treatment outcome was compared to non-injected (control) and human corneal fibroblasts (SFs)-injected corneas. Each group was comprised of at least 6 rats. Three weeks later, the rats were subjected to a visual function test in a Morris water maze. The corneas were harvested for immunofluorescence staining (IF) of keratocan, lumican, ALDH3A1, collagen 3A1, and α-smooth muscle actin (ASMA) and for synchrotron small-angle X-ray scattering (SAXS).

Results : The CSK injection was more effective in resolving the corneal haze than the SF- and non-treated corneas. The resolution was associated with 1.25x and 1.32x faster time and 1.93x and 1.65x shorter distances taken by the rats to swim to the platform when compared to the SF- and non-treated rats, respectively. From IF, we deduced that the superior haze resolution in the CSKs-injected corneas was due to the absence of collagen 3A1 and ASMA and the restoration of proteoglycans close to the state of a naïve cornea. In addition, SAXS revealed that the CSKs restored the collagen fiber organization (matrix order) to that of naïve corneas (p=0.961). In contrast, the SFs-injected and control corneas had a significantly lower matrix order than the naïve tissues (both p<0.001).

Conclusions : Injection of cultured CSKs improved visual functions and is a promising cell therapy for acute corneal haze. The CSKs restored the cornea’s collagen fiber organization and proteoglycans. Injection of SFs did not yield the same collagen fiber reorganization, resulting in poorer haze resolution.

This abstract was presented at the 2023 ARVO Annual Meeting, held in New Orleans, LA, April 23-27, 2023.

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