An injectable dextran/gelatin composite hydrogel loaded with exosomes for repair of rat intervertebral disc degeneration

doi:10.12307/2023.554

Abstract

Abstract: BACKGROUND: Exosomes can improve the inflammatory microenvironment of the degenerated intervertebral disc. However, direct injection of exosomes into the degenerated intervertebral disc at local concentrations and transient release cannot guarantee sustained effects, so the in vivo therapeutic effect of exosomes alone is limited.
OBJECTIVE: To observe the effect of exosome-loaded glucan/gelatin composite hydrogel on repairing rat degenerative intervertebral disc.
METHODS: Exosomes were extracted from bone marrow mesenchymal stem cells of rats. Exosome-loaded dextran/gelatin composite hydrogels were prepared by Schiff crosslinking reaction, and the microstructure and elastic modulus of the hydrogels were detected. Sixty Sprague-Dawley rats were randomly divided into five groups, with 12 rats in each group. Group A was the normal control. In groups B, C, D, and E, the intervertebral disc degeneration model was established by percutaneous puncture. PBS, exosomes, glucan/gelatin composite hydrogel and exosome-loaded glucan/gelatin composite hydrogel were injected into the intervertebral disc in turn. At postoperative 8 weeks, imaging, histopathology and western blot assay were performed.
RESULTS AND CONCLUSION: (1) The exosome-loaded glucan/gelatin composite hydrogels presented a three-dimensional porous structure with interconnected pores. The pore size was 50-200 μm, and the elastic modulus was (4.41±0.23) kPa, respectively. The exosomes could be released continuously and stably in vitro for more than 20 days. (2) At 8 weeks after operation, X-ray films exhibited that the intervertebral disc height index percentages of rats in groups B-E were lower than those in group A (P < 0.05). The intervertebral disc height index percentages of rats in groups C, D and E were higher than those in group B
(P < 0.05), of which group E was the most obvious. At 8 weeks after operation, MRI examination showed compared with group A, the tissue structure of nucleus pulposus in groups B-E was damaged to different degrees. Compared with group B, the tissue structure of nucleus pulposus in groups C-E was improved to varying degrees, among which group E had the most obvious improvement. (3) Hematoxylin-eosin staining and saffron O staining at 8 weeks after surgery exhibited that the structure of the nucleus pulposus in group B was severely damaged and only contained a small amount of collagen. Compared with group B, the structure of nucleus pulposus in groups C-E was improved, and the structure of nucleus pulposus in group E was finer, clearer, regular and had the most collagen content. (4) Western blot assay demonstrated that compared with group A, the expression levels of type II collagen and proteoglycan protein in groups B-E were decreased (P < 0.05). Compared with group B, the expression levels of type II collagen and proteoglycan protein in groups C-E were increased (P < 0.05), and those in group E were the most obvious. (5) It is concluded that the dextran/gelatin composite hydrogel loaded with exosomes injected into the degenerative intervertebral disc can restore the intervertebral disc height and intervertebral disc signal intensity, restore the tissue structure, and delay intervertebral disc degeneration to a certain extent.

Key words: intervertebral disc degeneration, hydrogel, exosome, composite hydrogel, animal experiment

CLC Number:

Pan Yujun, Shi Changjiang, Cao Sheng, Mu Huaizhao, Zhang Yilong, Wang Kaihua. An injectable dextran/gelatin composite hydrogel loaded with exosomes for repair of rat intervertebral disc degeneration[J]. Chinese Journal of Tissue Engineering Research, 2023, 27(34): 5477-5482.

Figures/Tables 7

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