Generation of Retinal Pigmented Epithelium-Like Cells from Pigmented Spheres Differentiated from Bone Marrow Stromal Cell-Derived Neurospheres

BACKGROUND: Retinal degeneration causes blindness, and cell replacement is a potential therapy. The purpose of this study is to formation of pigmented neurospheres in a simple medium, low-cost, high-performance manner over a short period of time while expressing markers of RPE cells and the activation of specific genes of the pigment cells. Also, these neurospheres have the ability to produce a monolayer of retinal pigment epithelium-like cells (RPELC) with the ability of photoreceptor outer segment phagocytosis. METHODS: BMSC were isolated from pigmented hooded male rats and were immunoreactive to BMSC markers, then converted into neurospheres, differentiated into pigmented spheres (PS), and characterized using Retinal pigment epithelium-specific 65 kDa protein (RPE65), Retinaldehyde-binding protein 1 (CRALBP) and orthodenticle homeobox 2 (OTX2) markers by immunocytochemistry, RT-PCR and RT-qPCR. The PS were harvested into RPELC. The functionality of RPELC was evaluated by phagocytosis of fluorescein-labeled photoreceptor outer segment. RESULTS: The BMSC immunophenotype was confirmed by immunostained for fibronectin, CD90, CD166 and CD44. These cells differentiated into osteogenic and lipogenic cells. The generated neurospheres were immunoreactive to nestin and stemness genes. The PS after 7–14 days were positive for RPE65 (92.76–100%), CRALBP (95.21–100%) and OTX2 (94.88–100%), and after 30 days RT-PCR, qPCR revealed increasing in gene expression. The PS formed a single layer of RPELC after cultivation and phagocyte photoreceptor outer segments. CONCLUSION: Bone marrow stromal stem cells can differentiate into functional retinal pigmented epithelium cells in a simple, low-cost, high-performancemanner over a short period of time. These cells due to expressing theRPELCgenes andmarkers can be used in cell replacement therapy for degenerative diseases including age-relatedmacular degeneration as well as retinitis pigmentosa.

Evaluation of Pain and Its Effect on Quality of Life and Functioning in Men with Spinal Cord Injury

BACKGROUND: Pain is one of the most important consequences of spinal cord injury (SCI). It may affect several aspects of life, especially the quality of life (QoL). Hence, this study was conducted to establish an understanding of pain and its correlates and effects on patients with SCI in our community. METHODS: In a cross-sectional study, 58 male veterans suffering from SCI were admitted to our center for a regular follow-up. Demographic and SCI-related descriptive information were gathered using a self-reported questionnaire. To evaluate the patients' pain quality and the effect of pain on daily life, a questionnaire in 3 parts of lumbar, cervical and shoulder pain was administered. EuroQoL questionnaire and General Health Questionnaire (GHQ) 12 were also used to assess the patients' QoL. RESULTS: The mean age of the participants was 45.91 +/- 6.69 with mean injury time of 25.54 +/- 5.91. forty-four patients (75.9%) reported pain, including lumbar pain (63%), cervical pain (39%) and shoulder pain (51%). The presence of pain was associated with lower QoL. Patients with lumbar pain reported a significant amount of pain affecting their daily life and this effect was higher in patients with lower GHQ score or anxiety/depressive disorder. CONCLUSIONS: Musculoskeletal pain, is a common complaint in veterans with SCI and is inversely associated with functioning and general health status. Lumbar and shoulder pain affects patient's daily living more than cervical pain.