Improvement of Photoreceptor Function Following Transplantation of NS-Derived RPE Cells into the Subretinal Space of an Animal (Rat) Model of Retinal Degeneration
Hamid Aboutaleb Kadkhodaeian1,2, Taki Tiraihi1*, Hamid Ahmadieh3, Hossein Ziaei3, Narsis Daftarian4 and Taher Taheri5
1Department of Anatomical Sciences, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran
2Department of Anatomical Sciences, Faculty of Medicine, Semnan University of Medical Sciences, Semnan, Iran
3Ophthalmic Research Center, Research Institute for Ophthalmology and Vision Science, Shahid Beheshti University of Medical Sciences, Tehran, Iran
4Ocular Tissue Engineering Research Center, Research Institute for Ophthalmology and Vision Science, Shahid Beheshti University of Medical Sciences, Tehran, Iran
5Shefa Neuroscience Research Center, Khatam-Alanbia Hospital, Rashid Yasemi Street, Tehran, Iran
*Corresponding Author: Taki Tiraih, Department of Anatomical Sciences, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran.
June 22, 2022; Published: July 19, 2022
Purpose: To investigate transplantation of retinal pigment epithelium (RPE) antigen (PSRA)-expressing pigmented spheres in an animal (rat) model of age-related macular degeneration (AMD) using sodium iodate to rescue and improve (i) a- and b-wave activities, (ii) alter outer nuclear layer thickness, and (iii) enhance cell number.
Materials and Methods: Male hooded rats (n = 65) were divided into five groups, two of which received sodium iodate and three of which did not. AMD was induced using retro-orbital sodium iodate injection. After 30 days, cells were injected into the subretinal space using a trans-scleral approach. For cell transplantation, rat bone marrow stromal stem cells were differentiated into neurospheres (NSs) and, after 7 days, into RPE cells. For tracking, differentiated cells were labeled with BrdU and then transplanted into the subretinal space. Photoreceptor function was evaluated by full-field electroretinography over the course of 7-90 days. The effects of transplanted cells on neurosensory retina and RPE layer were assessed using immunohistochemistry and cresyl violet staining at corresponding time points.
Results: Both the scotopic b-wave at an intensity of 0.01 cd.s/m2 and photopic a-wave at an intensity of 3.0 cd.s/m2 were affected. Significant differences between the test groups and relevant controls appeared at 60 days, but only for the scotopic assay and, at 90 days, for the photopic assay. Seven days after injection, light microscopy of IHC on paraffin sections in the transplanted group showed that PSRA cells had migrated and integrated into the host RPE layers. Further investigation using specific RPE cell protein RPE65 and ZO-1 revealed that these cells were able to express specific proteins as well. There was a statistically significant difference between the numbers of outer nuclear layer (ONL) cells and thicknesses in transplanted group. Significant differences between the test group and relevant controls appeared throughout the 7-90 day course in ONL cell count and through 14-90 days in ONL thickness. Ninety days after transplantation, the RPE layer and the neurosensory retinal layers were detectable. There was a surprising affinity between the PSRA cells and the host RPE layer.
Discussion: We demonstrated that PSRA migrated into the subretinal space and integrated into the host layer, expressing ZO-1, and RPE65 markers. Additionally, we showed that photoreceptor activity improved. The ONL was thicker, and ONL cell numbers were better preserved than in RPE-damaged rats that received only phosphate buffered saline (PBS).
Keywords: Photoreceptor Function; RPE; Subretinal Space; Sodium Iodate; Rat BMSCsc
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