Polydatin loaded collagen/heparin sulfate scaffold in the treatment of spinal cord injury in rats

doi:10.12307/2022.305

Abstract

Abstract: BACKGROUND: At present, the research on the neuroprotective effect of polydatin has gradually become a hot spot in the field of neurology. The research direction is mostly focused on ischemic cerebrovascular disease, and there are few studies on the application of spinal cord injury.
OBJECTIVE: With collagen/heparin sulfate scaffold as carrier, polydatin was applied to the injured spinal cord to observe the repair effect.
METHODS: (1) Collagen/heparin sulfate scaffolds and collagen/heparin sulfate scaffolds loaded with 0.5, 1, 1.5 mmol/L polydatin were prepared. The third generation of rat neural stem cells was seeded on four kinds of scaffolds, and the proliferation of cells was detected by CCK-8 assay. During inducing neural differentiation of neural stem cells, the expression levels of glial fibrillary acidic protein, Tuj-1 and Oligo were detected by immunofluorescence staining and RT-PCR. (2) The spinal cord injury model of adult male SD rats was established and divided into three groups. The model control group (n=10) was not implanted with any materials. The control group (n=10) was implanted with collagen/heparin sulfate scaffold, and the experimental group (n=10) was implanted with 1 mmol/L polydatin/collagen/heparin sulfate scaffold. Meanwhile, sham operation group (n=10) was set up. BBB score was used to test the motor function of the right hind limb within 8 weeks after operation. At 8 weeks after operation, the spinal cord tissues were taken for histological observation, immunohistochemical analysis, and western blot assay.
RESULTS AND CONCLUSION: (1) At 3 and 7 days of culture, the absorbance value of cell proliferation on 1 mmol/L polydatin/collagen/heparin sulfate scaffold was higher than that on the other three scaffolds (P < 0.05). (2) Immunofluorescence staining 7 days after induction showed that the expression of glial fibrillary acidic protein in 1 and 1.5 mmol/L polydatin/collagen/heparin sulfate scaffold groups were less than those in the other two collagen/heparin sulfate scaffold groups, but the expression levels of Oligo and Tuj-1 were more than those in the other two collagen/heparin sulfate scaffold groups. (3) RT-PCR results showed that the expression of glial fibrillary acidic protein mRNA in 1 and 1.5 mmol/L polydatin/collagen/heparin sulfate scaffold groups was lower than that in the other two collagen/heparin sulfate scaffold groups (P < 0.05). There was no significant difference in the expression of Tuj-1 mRNA among the four groups. The expression of Oligo mRNA in 1 mmol/L polydatin/collagen/heparin sulfate scaffold group was higher than that in the other three groups (P < 0.05). (4) Spinal cord injury repair experiments showed that the BBB score of the experimental group was always higher than that of the model control group and the control group 2-8 weeks after operation (P < 0.05). Hematoxylin-eosin staining showed that the spinal cord defects in the control group and the observation group were filled with scaffolds, and the space between the tissues was smaller than that in the model control group. The tissue continuity and space in the observation group were better than those in the control group. Immunohistochemical analysis and western blot assay showed that the expression of neurofilament-200 protein in the experimental group was higher than that in the control group and model control group (P < 0.05). The expression of glial fibrillary acidic protein in the experimental group was lower than that in the control group and model control group (P < 0.05). (5) It is concluded that polydatin loaded collagen/heparin sulfate scaffolds can promote the repair of spinal cord injury.

Key words: polydatin, collagen, heparin sulfate, scaffold, tissue engineering scaffold, biomaterials, spinal cord repair, spinal cord injury

CLC Number:

Liu Kang, Wang Yongping, Zuo Kebin, Yang Haitao, Jia Bin. Polydatin loaded collagen/heparin sulfate scaffold in the treatment of spinal cord injury in rats[J]. Chinese Journal of Tissue Engineering Research, 2022, 26(28): 4500-4506.

Figures/Tables 10

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