Adipose tissue derived extracellular vesicles promote nerve regeneration and functional recovery after sciatic nerve injury in rat
10.3760/cma.j.cn441206-20240626-00158
- VernacularTitle:脂肪组织来源细胞外囊泡促进大鼠坐骨神经损伤后的神经修复
- Author:
Siwen ZHANG
1
;
Jinsheng HUANG
;
Laijin LU
;
Nan ZHOU
Author Information
1. 郑州大学第一附属医院骨科，郑州 450000
- Publication Type:Journal Article
- Keywords:
Adipose tissue derived extracellular vesicles;
Sciatic nerve injury;
Axon regeneration;
Rat;
Nerve function recovery
- From:
Chinese Journal of Microsurgery
2025;48(2):179-186
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To study the effect of adipose tissue derived extracellular vesicles (AT-EVs) on nerve regeneration and functional reinnervation after sciatic nerve injury (SNI) in rat.Methods:From April 2023 to April 2024, the experiment was completed by the Department of Orthopaedics, the First Affiliated Hospital of Zhengzhou University in the key laboratory of the School of Medical Sciences, Zhengzhou University. AT-EVs were isolated and extracted from subcutaneous adipose tissue of 12 healthy male SD rats using ultracentrifugation. The morphology, size and distribution of AT-EVs were characterised via transmission electron microscopy (TEM) and nanoparticle tracking analysis (NTA). The effect of AT-EVs on the proliferation ability of pheochromocytoma cells of rat (PC12) and rat Schwann cells (RSC 96) were detected by cell proliferation assay. Twenty-four healthy male SD rats were randomly divided into 3 groups with 8 rats per group, including a sham group, a SNI group and an AT-EVs treatment group. Right sciatic nerves of rat models in sham group were exposed without injury. For the rat models in SNI group and AT-EVs treatment group, right sciatic nerves were injected with phosphate buffered solution (PBS) and AT-EVs under the epineurium of the nerve respectively following sciatic nerve crush. At 28 days after surgery, Sciatic function index (SFI) and pace analysis was performed. Then, rats in all groups were sacrificed, and bilateral gastrocnemius muscles were collected to measure the muscle wet weight ratio. The right sciatic nerves were removed for further exploration of nerve regeneration. The histopathological changes and axon arrangement of sciatic nerve were evaluated by Hematein Eosin (HE) staining and Masson staining. The regeneration efficiency of Myelin forming protein (MBP) and Neuronal marker [β-tubulin (Tuj-1)] were analysed by immunofluorescence staining of sciatic nerve. Statistical analysis was conducted using GraphPad Prism 8.0. P<0.05 were considered statistically significant. Results:The results of TEM and NTA revealed that AT-EVs exhibited a typical disc-shaped bilayer membrane structure with a predominant particle size of 184.5 nm. AT-EVs significantly enhanced the proliferation ability of PC12 and RSC96, and the difference was statistically significant ( P<0.05). At 28 days after surgery, the SFI and muscle wet weight ratio were higher in the AT-EVs treatment group compared to the SNI group, and the difference was statistically significant ( P<0.05). Axons in AT-EVs treatment group were arranged more closely and orderly than those in SNI group at 28 days after surgery, and there were less axon disintegration and vacuolation caused by injury. The immunofluorescence assay indicated that the MBP and Tuj-1 fluorescence intensity in AT-EVs treatment group was significantly higher than those in the SNI group, and the difference was statistically significant ( P<0.05). Conclusion:AT-EVs can enhance the proliferation ability of PC12 and RSC96 to promote axon regeneration and functional reinnervation following SNI.
