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.

Decrease in cavity size and oligodendrocyte cell death using Neurosphere-Derived oligodendrocyte-like cells in spinal cord contusion model

Background: Oligodendrocyte cell death is among the important features of spinal cord injury, which appears within 15 min and occurs intensely for 4 h after injury, in the rat spinal contusion model. Accordingly, the number of oligodendrocytes progressively reduced within 24 h after injury. Administration of oligodendrocyte-like cells [OLCs] into the lesion area is one of the approaches to counterbalance this condition

Methods: Bone marrow stromal cells were transdifferentiated into neurospheres and then into neural stem cells and later were differentiated into OLCs using triiodothyronine and transplanted into the spinal cord contusion rats. The postinjury functional recovery was explored and compared with the control group using Basso-Beattie-Bresnahan and narrow beam behavioral tests. At the end of 12th week, spinal cord segments T12-L1 were histomorphologically studied by immunohistochemistry

Results: Motor improvement was more obvious during 2nd to 4th weeks and got less prominent during 4th to 12th weeks. Histomorphometric findings indicated that cavity formation decreased in epicenter of transplantation area in experimental groups in comparison with the control groups

Conclusion: The findings obtained in the present study showed that OLC therapy is a potential approach in the treatment of spinal cord traumatic injuries

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.

Effects of transfer training on musculoskeletal pain in the caregiver of a stroke patient: a case report

The purpose of this case report was to evaluate the effectiveness of training of transfer techniques to a caregiver of a person who had suffered a stroke in decreasing musculoskeletal pain, depression and anxiety. This study adopted a single subject research design to evaluate the effectiveness of transfer-techniques training on musculoskeletal pain, depression, and anxiety in a 25-year-old female caregiver of a person with a stroke. The study was completed in four phases, including a baseline evaluation [1st and 3rd week], training [3rd, 5th and 7th week], post-training [9th week], and follow-up evaluation [11th week]. During the 1st week, demographic and descriptive information [such as age, time since diagnosis, cognition and independence of daily living] were collected from the stroke patient. Also, pain severity, anxiety and depression levels of the caregiver were evaluated. In weeks 3, 5 and 7, transfer training was undertaken. The patient was involved in the training with the caregiver under the supervision of an occupational therapist in their own home. The effectiveness of the training with regard to musculoskeletal pain and depression and anxiety levels of the caregiver was evaluated in the 5th, 9th and 11th weeks. The data were analyzed using a visual analysis of trends and levels. The results showed a decrease in pain severity, anxiety and depression during training and post-training. The changes continued during the follow-up stage. This study suggests promising results for the effectiveness of the transfer-techniques training and justifies further clinical trials. A larger trial is required to confirm the effectiveness of transfer training in improving pain management in caregivers of stroke survivors

Bone marrow stromal cell transdifferentiation into oligodendrocyte-like cells using triiodothyronine as a inducer with expression of platelet-derived growth factor ? as a maturity marker

Background: the present study investigated the functional maturity of oligodendrocyte derived from rat bone marrow stromal cells [BMSC]

Methods: the BMSC were isolated from female Sprague-Dawley rats and evaluated for different markers, such as fibronectin, CD106, CD90, Oct-4 and CD45. Transdifferentiation of OLC from BMSC was obtained by exposing the BMSC to DMSO and 1 [micro]M all-trans-retinoic acid during the preinduction stage and then induced by heregulin [HRG], platelet-derived growth factor AA [PDGFR-alpha], fibroblast growth factor and T3. The neuroprogenitor cells [NPC] were evaluated for nestin, neurofilament 68, neurofilament 160 and glial fibrillary acidic protein gene expression using immunocytochemistry. The OLC were assessed by immunocytochemistry for O4, oligo2, O1 and MBP marker and gene expression of PDGFR-alpha was examined by RT-PCR

Results: our results showed that the fibronectin, CD106, CD90, CD45 and Oct-4 were expressed after the fourth passage. Also, the yield of OLC differentiation was about 71% when using the O1, O4 and oligo2 markers. Likewise, the expression of PDGFR-alpha in pre-oligodendrocytes was noticed, while MBP expression was detected in oligodendrocyte after 6 days of the induction

Conclusion: the conclusion of the study showed that BMSC can be induced to transdifferentiate into mature OLC. Iran. Biomed. J. 17 [2]: 62-70, 2013

Microglial activation in rat experimental spinal cord injury model

The present study was designed to evaluate the secondary microglial activation processes after spinal cord injury [SCI]. A quantitative histological study was performed to determine ED-1 positive cells, glial cell density, and cavitation size in untreated SCI rats at days 1, 2, and 4, and weeks 1, 2, 3, and 4. The results of glial cell quantification along the 4900- Micro m long injured spinal cord showed a significant increase in glial cell density percentage at day 2 as compared to other days. Whereas the highest increase in ED-1 immunoreactive cells [monocyte/phagocyte marker in rats] was observed at day 2 [23.15%] post-injury. Evaluation of cavity percentage showed a significant difference between weeks 3 and 4 post-injury groups. This study provides a new insight into the multiphase immune response to SCI, including cellular inflammation, macrophages/microglia activation, glial cell density, and cavitation. Better understanding of the inflammatory processes associated with acute SCI would permit the development of better therapeutic strategies

Remyelination of demyelinated rat spinal cord model by transplanting neural stem cells

Demyelination of CNS axons occurs under pathological conditions such as multiple sclerosis and spinal cord injuries, but can be repaired by cell therapy. Within the CNS remyelination can be achieved by transplantation of neural stem cells [NSCs]. NSCs are self-renewing cells that maintain the capacity to differentiate into CNS-specific cell types and can differentiate into the three main neural phenotypes: astroglia, oligodendroglia and neurons. They may also replace or repair diseased CNS tissue. Bone marrow stromal cells [BMSCs] were aseptically isolated from the tibia and femurs of young adult Sprague Dawley rats. BMSCs were evaluated by fibronectin and CD31 markers. BMSC-derived NSCs were evaluated by nestin and NF-68. An ethidium bromide-induced demyelinated dorsal column lesion was produced in young adult rats. Transplanting NSCs derived-BMSCs into demyelinated lesions after 3 days in adult rat spinal cords was done. Three weeks after transplantation of NSCs, the spinal cords were processed to evaluate remyelination by Luxol fast blue staining. After passage 3, BMSCs were evaluated and the result, showed the percentage of immunoreactive cells to fibronectin [94.7 +/- 2.65], however BMSC-derived NSCs expressed nestin [86.15 +/- 0.64] and NF-68 [84.55 +/- 0.94] which correlated with fibronectin down regulation. Histologically, the lesions showed slightly irregular elongated areas and had an average length of 1336.36 +/- 39.43 microm. Transplanted NSCs were capable of eliciting remyelination. These data support the conclusion that transplantation of NSCs results in functional remyelination of a dorsal column lesion and have valuable applications in the treatment of neurodegenerative diseases such as spinal cord injuries