Preparation and evaluation of an improved model of contusion spinal cord injury in rats

doi:10.3877/cma.j.issn.2095-9141.2023.06.002

Abstract

Abstract:

Objective

To prepare a rat contusion SCI model using a novel spinal cord injury (SCI) impactor and evaluate the model, in order to lay the foundation for exploring the mechanism of neuropathic pain after SCI and early intervention and treatment.

Methods

One hundred adult male SD rats were randomly divided into Control group (n=20), Sham group (n=40) and SCI group (n=40). Rats in SCI group were exposed to T10 segments and the SCI model was established by modified Allen's method. Rats in Sham group were exposed to T10 segments without SCI, while rats in Control group were not intervened. Open field test (BBB score) was used to evaluate the motor function of rats 1 d before injury and 1, 3, 7, 14, 21 and 28 d after injury. The mechanical foot retraction threshold (MWT) of the hind limb was measured 1 d before injury and 14, 21 and 28 d after injury, and the histopathological changes of the spinal cord were detected by hematoxylin eosin (HE) staining to evaluate whether the SCI model was successfully constructed.

Results

(1) The BBB scores of three groups of rats at 1, 3, 7, 14, 21, and 28 d after injury were compared, and the differences were statistically significant (P<0.05). Among them, the BBB scores of the SCI group were lower than those of the Control group and the Sham group, and the differences were statistically significant (P<0.05). The BBB scores of the SCI group were all 0 points on the 1^st day after injury, and showed an upward trend at 3, 7, 14, 21, and 28 d after injury, and the difference between any two time points in pairwise comparison was statistically significant (P<0.05). (2) There was no statistically significant difference in rat hindlimb MWT among the three groups 1 d before injury (P>0.05), but there was a statistically significant difference at 14, 21, and 28 d after injury (P<0.05). Moreover, the rat hindlimb MWT of SCI group was significantly lower than that of the Control group and Sham group, and the difference was statistically significant (P<0.05). The rat hindlimb MWT of SCI group rats showed a trend of first decreasing and then rebounding at 1 d before injury and 14, 21, and 28 d after injury. Except for no statistically significant difference between 21 d after injury and 28 d after injury (P>0.05), there were statistically significant differences in hindlimb MWT at all other time points (P<0.05). (3) On the 14 d after injury, HE staining showed extensive infiltration of spinal cord glial cells. A large number of glial cells differentiated and proliferated 21 d after injury, and a large amount of glial fibers were generated 28 d after injury.

Conclusion

The contusion SCI model established by using the new percussion device has the advantages of digitalization, stability, standardization, strong operability and good repeatability, can provide a standardized model for the pathological mechanism of neuropathic pain formation after SCI and the study of targeted drugs.

Key words: Contusion spinal cord injury, Improved animal model, Neuropathic pain, Mechanical withdrawal threshold

Yehuang Chen, Kaiqin Chen, Liang Xue, Jianwu Wu, Yubei Huang, Liangfeng Wei, Bingxiang Zeng, Shousen Wang. Preparation and evaluation of an improved model of contusion spinal cord injury in rats[J]. Chinese Journal of Neurotraumatic Surgery(Electronic Edition), 2023, 09(06): 325-332.

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References 23

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组别	只数	伤后1 d	伤后3 d	伤后7 d	伤后14 d	伤后21 d	伤后28 d	F值	P值
Control组	5	20.60±0.89	20.60±0.89	20.60±0.89	20.60±0.89	20.60±0.55	20.60±0.55	<0.001	1.000
Sham组	10	20.20±0.92	20.20±1.40	20.00±1.49	20.50±0.85	20.70±0.67	20.80±0.63	2.978	0.128
SCI组	10	0.00±0.00^ab	0.40±0.10^ab	3.60±1.07^ab	10.50±1.08^ab	15.90±2.33^ab	18.70±1.57^ab	464.306	<0.001
F值		2527.148	1040.879	541.671	328.933	27.675	10.199
P值		<0.001	<0.001	<0.001	<0.001	<0.001	0.001

组别	只数	伤后1 d	伤后3 d	伤后7 d	伤后14 d	伤后21 d	伤后28 d	F值	P值
Control组	5	20.60±0.89	20.60±0.89	20.60±0.89	20.60±0.89	20.60±0.55	20.60±0.55	<0.001	1.000
Sham组	10	20.20±0.92	20.20±1.40	20.00±1.49	20.50±0.85	20.70±0.67	20.80±0.63	2.978	0.128
SCI组	10	0.00±0.00^ab	0.40±0.10^ab	3.60±1.07^ab	10.50±1.08^ab	15.90±2.33^ab	18.70±1.57^ab	464.306	<0.001
F值		2527.148	1040.879	541.671	328.933	27.675	10.199
P值		<0.001	<0.001	<0.001	<0.001	<0.001	0.001

组别	只数	伤前1 d	伤后14 d	伤后21 d	伤后28 d	F值	P值
对照组	5	17.00±2.74	17.00±2.74	18.00±2.74	17.00±2.74	0.146	0.930
假手术组	10	17.50±2.64	16.50±2.42	16.00±2.11	17.50±2.64	1.976	0.167
SCI组	10	16.50±2.42	7.60±0.97^abc	9.60±0.70^abcd	9.90±0.32^abcd	82.375	<0.001
F值		0.379	59.787	46.271	39.143
P值		0.689	<0.001	<0.001	<0.001

组别	只数	伤前1 d	伤后14 d	伤后21 d	伤后28 d	F值	P值
对照组	5	17.00±2.74	17.00±2.74	18.00±2.74	17.00±2.74	0.146	0.930
假手术组	10	17.50±2.64	16.50±2.42	16.00±2.11	17.50±2.64	1.976	0.167
SCI组	10	16.50±2.42	7.60±0.97^abc	9.60±0.70^abcd	9.90±0.32^abcd	82.375	<0.001
F值		0.379	59.787	46.271	39.143
P值		0.689	<0.001	<0.001	<0.001

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