George Tharion, (2011) Role of Bone Marrow Mesenchymal Stem Cells in Motor recovery of spinal cord injury in rats. Masters thesis, Christian Medical College, Vellore.
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Abstract
INTRODUCTION: Spinal cord injury is a major catastrophy with no solutions for cure, and realistic expectations and possibilities for spinal cord repair are being acknowledged recently. The incidence of spinal cord injuries is approximately 30 per million. The incidence of spinal cord injuries is highest in the age group of 16 to 30 years. More than 80% of spinal cord injury occurs in males. Unfortunately most of the victims are young, in the prime of life, left with permanent paralysis of limbs, bladder, bowel and sexual functions. This results in far reaching social, vocational and psychological consequences. Prevention of complications, stabilisation of spine and active rehabilitation and long term follow up constitute the mainstay of the management of persons with spinal cord injury. Early rehabilitation in an organised multidisciplinary SCI team has been shown to be beneficial with lower mortality, decreased pressure sores, and slightly greater chance of neurological recovery and shorter lengths of stay in the hospital. Surgical stabilisation helps in early mobilisation, but there are no significant differences in neurological level changes between early surgery, late surgery and no surgery. There was no evidence that routine early surgical intervention or decompression improved neurological outcomes. The national spinal cord injury study NASCIS trials I, II, and III on early steroids to limit cellular damage from secondary injury processes suggested this to be part of the early management of SCI. The conclusions of this study have been challenged due to a variety of reasons. AIM OF THE STUDY: To study the effect of Bone marrow Mesenchymal Stem Cell transplantation for motor recovery in rat models of spinal cord injury. OBJECTIVES: 1. Bone marrow mesenchymal stem cell (MSC)culture and characterization. 2. Transdifferentiation of MSC to neuronal cells. 3. MSC transplantation to injured spinal cord segment of rat models. 4. Outcome assessment of transplantation by BBB scale, and EMG. MATERIALS & METHODS: All experiments were conducted with the approval from Institutional Review Board and the Animal Ethics Committee. Materials required: Cell culture facilities: Phosphate buffered saline (PBS), Gibco,Catalog no: 10010. RosetteSep antibody cocktail, Stem Cell Technologies Inc, catalog. No: 15168. Ficoll-Paque, GE Healthcare Bio-Sciences, catalog .no: 17-1440-03. T75 culture flask, Greiner. Alpha - Minimum essential medium, Gibco, Fetal bovine serum, Gibco, catalog.no: 10082. L-Glutamine, Gibco, Catalog.no: 25030. 0.25%Trypsin-EDTA, Gibco, Catalog.No: 25200. Dulbaco modified eagle medium, Gibco,catalog.No: 11995. DiMethylSulphoxide, DMSO Sigma, Catalog no: C6295 Butylated hydroxyanisole (BHA), ACROS. Potassium chloride (Kcl) , Qualigens. Valproic acid, sigma, catalog. No:P4543. Forskolin, Sigma. Hydrocortisone, Sigma, catalog.No: H6909. Insulin, Sigma, catalog.No: I9278. bFGF, Invitrogen, catalog.no: 13256-029. 30 Hanks Balanced salt solution (HBSS), Gibco, catalog no: 14175. 100U/mL penicillin, 100μg/ml streptomycin and 25ng/ml of amphotericin-B. Centrifuge, REMI. Laminar flow hood, Kartos International. CO2 incubator, Thermo Electron Corporation. Inverted phase-contrast microscope. Animal experiments: Albio Wistar rats, Animal house with air-conditioned facilities, Operation theatre for anaesthesia and surgery. Ketamine. Xylazine. Operating microscope. Surgical instruments: Bone rongeur. Surgical scissors. Equipment for drop weight device and transducers to record forces. Transcranial cortical stimulator. EMG recorder. Gait analysis set up for rats and video recorder for BBB score. 3 D Injection device for cell transplantation. CONCLUSIONS: Spinal cord injury results in permanent paralysis, loss of bowel and bladder control and other autonomic functions. Despite extensive research, there is no treatment available for this condition. This study demonstrated the role of bone marrow mesenchymal stem cells in spinal cord injury in rat models. 1. Bone MarrowMesenchymal Stem Cells can be cultured from the bone marrow sample and can be characterised to be used for transplantation. 2. Bone marrow Mesenchymal Stem Cells can be trans differentiated into neural cells in vitro using appropriate inducing media. However this observation needs further confirmation. 3. Drop weight model of spinal cord injury gives a consistent comparable contusion model for research studies. 4. Transcranial stimulation can be used to elicit motor evoked potential to demonstrate functional continuity of corticospinal tract in rat models of spinal cord injury. 5. Transplantation of bone marrow stromal stem cells can improve the motor out come in rat models of spinal cord injury as measured by BBB motor score and electrophysiological studies.
| Item Type: | Thesis (Masters) |
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| Uncontrolled Keywords: | Bone marrow mesenchymal stem cells, motor recovery, spinal cord injury, rats. |
| Subjects: | MEDICAL > Physical Medicine and Rehabilitation |
| Depositing User: | Subramani R |
| Date Deposited: | 23 Mar 2020 14:32 |
| Last Modified: | 23 Mar 2020 14:32 |
| URI: | http://repository-tnmgrmu.ac.in/id/eprint/12411 |
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