An experimental study to test the efficacy of Mesenchymal Stem Cells in reducing corneal scarring in an ex-vivo organ culture model.

In this study, we evaluated the effect of placenta-derived Mesenchymal Stem Cells (MSCs) versus placebo in improving corneal transparency following experimental injury in an ex-vivo organ culture model of post-mortem human corneas. We also compared the influence of MSCs on the basic histopathology of the cornea and the immunohistochemistry markers of fibrotic corneal scarring. Mesenchymal Stem Cells extracted from the placenta were isolated and expanded in-vitro. Five pairs of post-mortem human corneas harvested for the corneal transplant of equal grade were included in the study. Corneas of the same pair were randomly assigned to either the case arm or the control arm. All corneas underwent a standardized superficial keratectomy, 4 mm in diameter. The case and control arm corneas received an intrastromal injection of MSCs and placebo respectively. The corneal button was maintained in an organ culture system for 28 days under the standard protocol. Laser light was passed through the corneas mounted on a self-styled modified artificial anterior chamber. Image analysis was used to quantify corneal transparency. Haematoxylin & Eosin staining and Immunohistochemistry was done for Alpha SMA (Smooth Muscle Actin). Laser scatter measurements were measured using Image Analysis (Image J Software). The difference in the mean of Full-Width Half Maximum (FWHM), Max intensity and Red pixel intensity between the cases and the controls was 101.5, 16.3 and 11.4 respectively which was found to be statistically significant (P < 0.05). Histopathology showed a disorganized Bowman's layer in the controls as compared to the cases. Alpha Smooth Muscle Actin at the injury site stained 3 + in all controls as compared to 1 + in the cases, showing a statistically significant difference (p = 0.005). Based on our findings, we consider that placenta-derived Mesenchymal Stem Cells can alter evolving corneal scarring into a more favourable outcome with better corneal transparency and lesser fibrotic corneal scarring.

Objective quantification of corneal haziness using anterior segment optical coherence tomography.

PURPOSE: To quantify normal corneal transparency by anterior segment optical coherence tomography (AS-OCT) by measuring the average pixel intensity. To analyze the variation in the average pixel intensity in mild and severe grades of corneal opacities. METHODS: This is an observational, cross-sectional study of 38 eyes from 19 patients with mild or severe grades of corneal opacities greater than 3 mm and a normal contralateral cornea. AS-OCT was performed centered on the opacity with a 3 mm cruciate protocol. A similar image is taken of the contralateral clear cornea in the same quadrant. The average pixel intensity was calculated in a standardized manner using MATLAB software. RESULT: The average pixel intensity of the normal cornea was 99.6 ± 10.9 [standard deviation (SD)]. The average pixel intensity of the mild and severe corneal opacities was 115.5 ± 9.1 and 141.1 ± 10.3, respectively. The differences were statistically significant. CONCLUSIONS: AS-OCT images can be used to quantify corneal transparency. Average pixel intensity is a measure that varies significantly with varying corneal opacification.