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Modeling of spinal cord injury in rats. Cell-mediated gene therapy, epidural stimulation
Russia (TaMSA-Tatarstan) - Kazan State Medical University, Kazan
Department of topographical anatomy and operative surgery
Bashirov Farid Vagizovich
Fadeev Filip Olegovich
English, Russian, Turkish
Type of Research Project
- Basic science
What is the background of the project?
The work on spinal cord injury in rats began at the department in 2014, at first it was about overcoming the consequences of spinal cord injury using only the gene therapy. But gene therapy alone is effective only first 3 weeks and this condition requires non-stop therapy including all standard procedures. It is well-known that epidural stimulation shows good effect on motor function restoration after SCI and we thought what if we try to combine both methods. And as a result, we saw good outcomes. Both of these methods work in synergy and bring better morphofunctional restoration effects.
What is the aim of the project?
The aim of the project is to prove the reliability of the model and to prove the efficacy of combined electro- and gene therapy.
What techniques and methods are used?
All surgical procedures were performed under aseptic conditions and were divided into three stages (1) Implantation of stimulating electrodes. This stage included an installation of a 12-pin headplug (Omnetics Connector Corporation, USA) and teflon-coated wire electrodes (AS632, Cooner Wire) to connect an EES stimulator to electrodes on the headplug. Stimulating electrodes were placed at C5 and at L2 before SCI at Th8–Th9 level. After the local laminectomy at C5 and L2 levels, the electrodes were fixed to the dura using 9.0 suture. The wires of the headplug were routed under the skin. In previous studies with EES on animals, stimulating electrodes have been implanted at various time points before, during or after SCI. In the current study, we implanted stimulating electrodes before SCI to minimize potential surgical complications affecting evaluation of locomotor recovery post traumatic spinal cord, avoid losing experimental animals during the electrode implantation surgery, and include only animals with successful headplug implantation resulting in muscle contractions. (2) Spinal cord injury (SCI). Contusion SCI was performed a week after implantation of electrodes in a standard way as described previously. A mid-dorsal skin incision was made from vertebrae Th7 to Th10, tendons and muscles were removed from the spinous processes and vertebral arches and laminectomy was performed at the Th8–Th9 vertebral level. Contusion injury of spinal cord was induced using an impact rod (weight 10 g, diameter 2 mm) centered above Th8 and dropped from a height of 25 mm onto the exposed dura mater of the spinal cord, which inflicts a moderate contusion injury. Before closing the wound two surgeons verified the contusion injury which was accompanied by the appearance of hematoma at the site exposed to the impact rod. (3) Gene therapy (xenotransplantation of genetically modified human UCB-MC). A deep level of anesthesia was maintained for four hours after modeling of SCI (supplemental doses of Zoletil administered, i.p. as needed). While rats were kept under anesthesia, their body temperature was maintained at 37-38°С with a heating pad. After 4 hours, animals underwent a laminectomy at the level of L4-L5. Intrathecal injection of 20 μl of solution was made using a 25-μl Hamilton syringe and a 33 gauge needle. Five animals with implanted stimulating electrodes received 2×106 human UCB-MC+Ad5-VEGF-GDNF-NCAM in 20 μl of saline prepared as described above. Ten rats after implantation of stimulating electrodes were injected with 20 μl of saline.
What is the role of the student?
- The tasks of the student will be performed on his/her own
- The tasks will be done under supervision
What are the tasks expected to be accomplished by the student?
Presentations according to topic, mostly to understand the main purposes of this work (what is spinal cord injury, what is the meaning to the humanity, the statistics; what is gene therapy; what is epidural stimulation in model of spinal cord injury), mastering some surgical procedures such as rules of aseptics and antiseptics, performing surgical approaches in rats models, basic knowledge of lab animals anatomy, mastering the LabChart software.
Will there be any theoretical teaching provided (preliminary readings, lectures, courses, seminars etc)
Preliminary reading will be provided for the students. Mostly about the project itself, related articles, basics of microsurgery. Frequency and literature load depends on a level of training of incoming students.
What is expected from the student at the end of the research exchange? What will be the general outcome of the student?
- No specific outcome is expected
What skills are required of the student? Is there any special knowledge or a certain level of studies needed?
Nothing specific, better be interested in surgery.
Are there any legal limitations in the student’s involvement
Type of students accepted
This project accepts: - Medical students - Graduated students (less than 6 months)
- Andrei A. Izmailov et al. “Spinal Cord Molecular and Cellular Changes Induced by Adenoviral Vector- and Cell-Mediated Triple Gene Therapy after Severe Contusion”; Front. Pharmacol.; 13 November 2017 https://doi.org/10.3389/fphar.2017.00813 Rustem Robertovich Islamov et al. “Evaluation of direct and cell-mediated triple-gene therapy in spinal cord injury in rats”; Brain Research Bulletin; Volume 132;June 2017; Pages 44-52 https://www.sciencedirect.com/science/article/abs/pii/S0361923017300953?via%3Dihub
- Rustem Robertovich Islamov et al. “Evaluation of direct and cell-mediated triple-gene therapy in spinal cord injury in rats”; Brain Research Bulletin; Volume 132;June 2017; Pages 44-52 https://www.sciencedirect.com/science/article/abs/pii/S0361923017300953?via%3Dihub
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