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Objective: Spinal cord injury is a common and frequently-occurring disease, can cause irreversible below the level of injury feeling, movement and sphincter dysfunction, clinical treatment is not satisfactory, the medical field has been hot and difficult. Intraperitoneal injection of the experimental application has a protective effect on spinal cord injury safflower flavin joint MSCs transplantation in the treatment of spinal cord injury model to explore the safflower flavin intraperitoneal injection of MSCs transplantation for spinal cord injury repair mechanisms, and treatment for spinal cord injury clinical effect to find a feasible way. Methods: adherent separation isolated and purified rat of MSCs observed morphological characteristics of primary and passaged MSCs, growth characteristics, and its feasibility as seed cells using flow cytometry analysis of the technical appraisal MSCs surface protein expression. Carrying the GFP gene adenovirus infection MSCs, fluorescent microscope the MSCs transfected situation and calculate the transfection efficiency, observe GFP-MSCs morphological characteristics, growth characteristics of GFP-labeled MSCs biological traits. 55 adult SD rats, male and female, depending on the approach, were randomly divided into five groups: A group of (safflower Flavin intraperitoneal injection of MSCs transplantation group), group B (normal saline intraperitoneal injection of MSCs transplantation group), C group (safflower flavin intraperitoneal injection of PBS group) and Group D (intraperitoneal saline PBS group), more than 10 in each group; E group for the supplement group, 15. Modified Allen T7 plane production model of spinal cord injury. After the success of the model, A group of MSCs (GFP-MSCs) and damage local real-time group B directly after transplantation mark of 6 × 106/60μl, C group and D group was injected with an equal volume of PBS. Group A and group C by intraperitoneal injection of 40mg/kg dose of safflower Flavin, B and D groups to the same dose of saline, 1 day, shared 2w. During the experiment, the animals of each group died from Group E randomly selected supplement, to resume calculating model animal feeding processing time, to ensure that in each group the same as the number of experimental animals. Postoperative 1w, 2w, 3w and 4w using the BBB score hindlimb locomotor function after cardiac perfusion harvested production of frozen sections of fluorescence microscope observation transplanted MSCs; the HE staining histological changes; Immunohistochemical detection of GFAP, NeuN and NGF expression. Results: 1, adherence separation separation purified adherent cell morphology, growth characteristics with MSCs consistent flow cytometry to detect cell surface protein expression: CD45-positive rate of 14.47%, 89.26% CD44 positive rate, CD29-positive rate of 93.27 suggesting that cells obtained is of MSCs; carrying the GFP gene adenovirus infection MSCs fluorescence microscopy observations confirm that GFP-labeled MSCs in successful the adenovirus transfection efficiency (93% ± 2%), transfected MSCs no significant changes in morphology, growth characteristics; 3, GFP-MSCs transplantation under a fluorescence microscope, various time points 1w, 2w, 3w and 4w frozen tissue slices can be observed in bright green fluorescence of GFP-MSCs issued to prove transplantation of MSCs can survive in the injured spinal cord in the rat; 4, after 2w-4W, safflower flavin intraperitoneal injection of MSCs transplantation alone can raise the the rat BBB score score, both associated with the role of more , the difference was statistically significant (P lt; 0.05), the postoperative 1w, each group rated the difference is not significant (P gt; 0.05); postoperative 1w, 2w, 3w and 4w point in time frozen tissue slices HE staining showed that safflower flavin intraperitoneal injection of MSCs transplantation alone can improve the nerve tissue lesions situation, reducing the formation of the cavity, and the degree of improvement and the BBB rating scores, both associated with the cytopathic effect is more obvious; 6 Immunohistochemical detection of groups GFAP, NeuN and NGF expression increased with the extension of time after the same point in time the same indicator group A, group B and group C expression levels were significantly higher in group D, significant differences (P lt; 0.05), where the expression levels of the group A than in group B and group C, the difference was significant (P lt; 0.05), the level of expression of group B than in group C, the difference was significant (P lt ; 0.05). Conclusion: 1, adherence separation is a better the MSCs separation method, received the rapid growth of adherent cells, vitality, flow cytometry analysis technology proved to MSCs, can be used as seed cells for tissue engineering; 2 application carrying the GFP gene adenovirus infection of MSCs, GFP transfection efficiency, no significant effect on the biological characteristics of MSCs labeled MSCs can survive in the rat spinal cord, display, GFP is a better marker of MSCs method ; 3, safflower flavin intraperitoneal injection or the MSCs transplantation able to enhance the secretion of NGF alone, prompting the differentiation of stem cells into nerve cells and glial cells direction conducive to the repair of damaged nerve tissue, both combined with a synergistic effect.
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