Purpose: To examine the protection of a single intravitreal injection of autologous bone Marrow Mesenchymal stem cells (MSCs) in patients with advanced retinitis pigmentosa (RP). in fibrosis; however, intravitreal injections of the two other patients’ cells into the mouse vitreous did not generate any fibrous tissue. Conclusion: Intravitreal injection of autologous bone marrow MSCs into patients’ eyes with advanced RP does not meet safety standards. Major side effects of this therapy can include fibrosis and TRD. We propose thorough evaluation of MSCs prior to transplantation by intravitreal injection in the laboratory animals.\ Animal Studies Adult male C57/BL6 mice and B6Nude mice (20C25 g) were housed under light- and temperature-controlled conditions. All experiments were conducted in compliance with the Association for Research in Vision and Ophthalmology (ARVO) Statement for the Use of Animals in Ophthalmic and Vision Research. The experimental protocol was approved by the Institutional Animal Care and Use Committee of Royan Institute. Procured MSCs obtained from the RP patients, were injected into the mice vitreous cavities and the corresponding eyes were subjected to routine histopathologic and immunohistochemical studies. For immunofluorescence analysis, the sections were permeabilized with 0.3% Triton X-100 in phosphate-buffered saline (PBS) and blocked with normal secondary host serum. The sections were stained overnight with primary antibodies against specific anti-human Thy1 Ac2-26 (Millipore, CBL415) and glial fibrillary acidic protein (GFAP; Sigma-Aldrich G3893). The stained sections had been examined having a fluorescent microscope (Olympus, IX71, Japan) that got a DP72 camera pursuing treatment with supplementary antibodies: Goat anti-mouse Alexa-fluor 568 Ac2-26 (Molecular Probes) or goat anti-mouse FITC (Sigma-Aldrich). Nuclei Ac2-26 had been stained with 4′,6-diamidino-2-phenylindole (DAPI). Several sections had been stained with hematoxylin and eosin (H and E). Outcomes Clinical Assessments MSCs useful for transplantation had been positive at passing one for Compact disc90 ( 90%), Compact disc105 ( 90%), Compact disc73 ( 90%) and Compact disc44 ( 60%). These MSCs demonstrated low manifestation of Compact disc11b ( 30%) and minor double manifestation of Compact disc45/34 ( 10%) [Shape 1]. Open up in another window Shape 1 Movement cytometry of bone tissue marrow-mesenchymal stem cells (MSCs) of individuals with retinitis pigmentosa (RP) at major culture. P, individual. 1 day after intravitreal transplantation of 106 MSCs per 0.1 ml, ocular examination revealed zero signals of any kind of significant intraocular increase or inflammation in IOP. Through the postoperative program, we noticed no symptoms of adverse occasions in individuals P1 and P2 at fourteen days after intravitreal shot from the MSCs [Shape ?[Shape2a2aCc]. These individuals reported visible improvement 14 days after intravitreal shot from the MSCs which persisted up Rabbit polyclonal to ZKSCAN4 to three months. Multifocal ERG performed at weeks 3, 6 and 12 exposed no significant modification set alongside the baseline mfERG. Baseline OCT demonstrated severe thinning from the macular middle, marked atrophy from the external retina, photoreceptor reduction, severe reduced amount of Ac2-26 the peripapillary nerve dietary fiber coating and central choroidal width. An evaluation of OCT results before with different time factors following the MSCs shot did not display any significant variations with regards to central macular width (CMT), peripapillary nerve dietary fiber layer width and central choroidal width (CCT) [Shape ?[Shape2a2aCc]. Baseline FAF demonstrated diffuse hypoautofluorescence without obvious modification following the shot [Shape ?[Shape2d2dCf]. FA revealed window defects and abnormal visibility of the choriocapillaris secondary to severe RPE atrophy in addition to extensive areas of hypofluorescence due to loss of the choriocapillaris. There was no significant leakage observed before and 3 months after the injection [Figure ?[Figure2g2g and ?andhh]. Open in a separate window Figure 2 Enhanced depth imaging-optical coherence tomography (EDI-OCT) (a-c), Ac2-26 auto-fluorescence (FAF) (d-f), and fluorescein angiography (FA) (g-h) images of the left eye of patient P2. (a) Prior to the cell injection, there was a severe reduction in the central macular and.