The effects of punctured nucleus pulposus on lumbar radicular pain in rats: a behavioral and immunohistochemical study

Laboratory investigation

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Object

Application of the nucleus pulposus (NP) to the nerve root has been developed as a model of lumbar radicular pain. The relationship between disc degeneration and the induction of radicular pain, however, has not yet been fully explored. The authors of this study investigated pain-related behaviors and expression of tumor necrosis factor–α (TNF-α) in the dorsal root ganglion (DRG) to evaluate the effects of punctured NP on lumbar radiculopathy.

Methods

An anular needle puncture model of intervertebral disc degeneration in a rat tail was established. Normal and previously punctured NP tissues were obtained and placed on the L-5 nerve root following a hemilaminectomy. Behavioral tests including assessment of motor function, mechanical threshold, and thermal withdrawal latency were performed before and after surgery. The TNF-α immunoreactivity in L-5 DRG specimens was examined through immunohistochemical study.

Results

The punctured discs showed significant degeneration 2 weeks after intervention. Application of both normal and punctured NP induced mechanical hyperalgesia in the ipsilateral paw for 10 days after surgery, but hyperalgesia was more severe in the punctured NP group. No statistically significant within-group changes in thermal withdrawal latency over time were found. A significant increase in the expression of TNF-α–positive neurons in DRG specimens was observed in both NP graft groups.

Conclusions

Needle puncture led to degenerative changes in the rat tail disc, and the degenerated NP enhanced mechanical hyperalgesia induced by application of the NP to the lumbar nerve root. This model of disc degeneration and lumbar radicular pain is appropriate for evaluating the efficacy of biological treatments for degenerative disc diseases.

Abbreviations used in this paper: DRG = dorsal root ganglion; IL = interleukin; NP = nucleus pulposus; TNF-α = tumor necrosis factor–α.

Article Information

Address correspondence to: Zhao-Min Zheng, M.D., Ph.D., Department of Spine Surgery, The First Affiliated Hospital, Sun Yat-sen University, 58 Zhongshan Road 2, Guangzhou, 510080, Guangdong, China. email:zhengzm1@163.com.

© AANS, except where prohibited by US copyright law.

Headings

Figures

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    Representative T2-weighted, sagittal MR images of the coccygeal spine obtained in rats from the Control (left) and P-NP (right) groups. Note the significant signal change at the operated levels (arrows) compared with the same levels in the tail in which sham surgery was performed (arrowheads).

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    Photomicrographs of midsagittal sections of rat tail discs. Discs from animals in the Control (A) and Sham Surgery (B) groups demonstrated a rounded NP, well-organized collagen lamellae, and a well-defined border between the anulus fibrosus and the NP. In the punctured disc (C), the amount of NP was decreased, the anulus fibrosus was disorganized, and the border was interrupted. H & E, original magnification × 4.

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    Graphs showing the mechanical withdrawal threshold of the rats' ipsilateral (left) and contralateral (right) hind paws. Results are expressed as the means ± SEMs of the 50% withdrawal threshold. NC = Control Group (naïve control); N-NP = N-NP Group (normal NP); P-NP = P-NP Group (needle-punctured NP); Sham = Sham Surgery Group. * Statistically significant difference between presurgical and postsurgical values (any group). # Statistically significant difference between the N-NP and P-NP groups.

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    Graphs showing the thermal withdrawal latency in testing of the rats' ipsilateral (left) and contralateral (right) hind paws. Results are expressed as the means ± SEMs of the 50% withdrawal threshold. * Statistically significant difference between presurgical and postsurgical values.

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    Photomicrographs showing the results of immunohistochemical staining of L-5 DRG specimens. Specimens from the N-NP (A) and P-NP (B) groups show a strong immunoreactivity compared with those from the Control (C) and Sham Surgery (D) groups. Bar = 50 μm.

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    Graphic illustration of the number of TNF-α positive cells in sections obtained at Days 4 and 14 after surgery. Results are expressed as means ± SEMs. The number of positive cells was significantly higher in the N-NP and P-NP groups compared with the Control and Sham Surgery groups. No statistically significant differences were observed between the N-NP and P-NP groups. * p < 0.05 versus Control Group. # p < 0.05 versus Sham Surgery Group.

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