A metabolic profiling analysis of degenerative intervertebral disc disease in a rabbit model via GC/TOF-MS

Abstract Background: To establish an analytical method for studying intervertebral disc metabolomics based on gas chromatography/time-of-flight mass spectrometry (GC/TOF-MS). Methods: To establish a rabbit model of intervertebral disc degeneration, the intervertebral discs of six New Zealand white rabbits were punctured. GC/TOF-MS was applied to analyze degenerated discs of the six model group animals and six similar New Zealand white rabbits considered as the control group. Pattern recognition and nonparametric test analyses were utilized to evaluate the data as well as screen for and identify significant biomarkers. Metabolites and metabolic pathways relevant to associated pathological processes were studied. Results: We established a rabbit model of intervertebral disc degeneration and an orthogonal partial least squares-discriminant analysis (OPLS-DA) disease-distinguishing model of intervertebral disc degeneration for contrast against the typical physiological state of intervertebral discs. Expression of six metabolites in degenerated and normal intervertebral discs of New Zealand white rabbits was evaluated. Levels of four metabolites expressed in the model group were significantly higher than in the control group, while two other metabolites were expressed at significantly lower levels in the model group as compared to the control group. Conclusions: This paper demonstrates that metabolic profiling of both degenerated and normal intervertebral rabbit discs is a feasible method of metabolomic analysis. Via in-depth filtering of characteristic metabolites, we found a high correlation between metabolic variations and intervertebral disc degeneration. Further studies on endogenous metabolites and pathways involved in intervertebral disc degeneration will provide a better understanding of associated molecular mechanisms and lay foundations for effective clinical treatment.

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Efficacy of Platelet-Rich Plasma Containing Xenogenic Adipose Tissue-Derived Stromal Cells on Restoring Intervertebral Disc Degeneration: A Preclinical Study in a Rabbit Model

Pain Research and Management ◽

10.1155/2019/6372356 ◽

2019 ◽

Vol 2019 ◽

pp. 1-7

Author(s):

Chao Ma ◽

Ran Wang ◽

Dingliang Zhao ◽

Naikun Wang ◽

Ying Han ◽

...

Keyword(s):

Adipose Tissue ◽

Intervertebral Disc ◽

Disc Degeneration ◽

Stromal Cells ◽

Signal Intensity ◽

Intervertebral Disc Degeneration ◽

Platelet Rich Plasma ◽

Rabbit Model ◽

Control Group ◽

Arterial Blood

Objective. Platelet-rich plasma (PRP) containing multiple growth factors is a promising strategy for disc degeneration. Thus, this study hypothesizes that the combination of PRP and adipose tissue-derived stromal cells (ADSCs) may repair degenerative disc more effectively than using each one of them alone. Methods. The model of early intervertebral disc degeneration was induced by annular puncture in the New Zealand rabbit. Autologous PRP was extracted from fresh arterial blood by using two centrifugation techniques. ADSC was offered by the Center for Clinic Stem Cell Research. Four weeks after the first experiment, PRP or ADSCs or a combination of PRP and ADSCs was injected into the punctured intervertebral disc. Four weeks later, disc height and signal intensity on T2-weighted magnetic resonance imaging (MRI) were assessed. Results. One month after puncture, we detected relatively narrow discs and lower signal intensity in MRI T2-weighted images. At four weeks after injection, the PRP-ADSC group statistically significantly restored discs, compared with PRP, ADSCs, or negative control group. Conclusions. The combination of PRP and ADSCs shows an effective potential to restore degenerated intervertebral discs in the rabbit.

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Histomorphometric and collagen fibers organization in annular punctured induced intervertebral disc degeneration in rabbit model: restorative and antioxidant role of Brassica oleracea

Advances in Traditional Medicine ◽

10.1007/s13596-021-00560-z ◽

2021 ◽

Author(s):

B. Ogunlade ◽

S. A. Adelakun ◽

B. K. Akinola ◽

O. A. Adedotun

Keyword(s):

Intervertebral Disc ◽

Brassica Oleracea ◽

Disc Degeneration ◽

Intervertebral Disc Degeneration ◽

Rabbit Model ◽

Collagen Fibers

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Transcriptomics Study to Determine the Molecular Mechanism by which sIL-13Rα2-Fc Inhibits Caudal Intervertebral Disc Degeneration in Rats

BioMed Research International ◽

10.1155/2020/7645989 ◽

2020 ◽

Vol 2020 ◽

pp. 1-13

Author(s):

Xin Wang ◽

Jianshi Tan ◽

Junhao Sun ◽

Pengzhong Fang ◽

Jinlei Chen ◽

...

Keyword(s):

Intervertebral Disc ◽

Disc Degeneration ◽

Intervertebral Disc Degeneration ◽

Type I Collagen ◽

Type Ii Collagen ◽

Type I ◽

Model Group ◽

Expression Levels ◽

Protein Levels ◽

Collagen Protein

Background. Intervertebral disc degeneration is related to tissue fibrosis. ADAMTS can degrade the important components of the ECM during the process of intervertebral disc degeneration, ultimately resulting in the loss of intervertebral disc function. sIL-13Rα2-Fc can inhibit fibrosis and slow down the degeneration process, but the mechanism involved remains unclear. Objective. To determine the mechanism by which sIL-13Rα2-Fc inhibits ECM degradation and reduces intervertebral disc tissue fibrosis using a transcriptomics analysis. Methods. A rat model of caudal intervertebral disc degeneration was established, and Sirius red staining was used to observe the pathological changes in the caudal intervertebral disc. Transcriptome sequencing was employed to assess the gene expression profiles of the intervertebral disc tissues in the model group and the sIL-13Rα2-Fc-treated group. Differentially expressed genes were identified and analyzed using GO annotation and KEGG pathway analyses. Real-time fluorescence quantitative PCR was used to verify the expression levels of candidate genes. The levels of GAG and HA were quantitatively assessed by ELISA, and the levels of collagen I and collagen II were analyzed by western blotting. Results. Sirius red staining showed that in the model group, the annulus fibrosus was disordered, the number of breaks increased, and the type I collagen protein levels increased, whereas in the sIL-13Rα2-Fc group, the annulus fibrosus was ordered, the number of breaks decreased, and the type II collagen protein levels increased. In comparison with the model group, we identified 58 differentially expressed genes in the sIL-13Rα2-Fc group, and these were involved in 35 signaling pathways. Compared with those in the model group, the mRNA expression levels of Rnux1, Sod2, and Tnfaip6 in the IL-13Rα2-Fc group were upregulated, and the mRNA expression levels of Aldh3a1, Galnt3, Fgf1, Celsr1, and Adamts8 were downregulated; these results were verified by real-time fluorescence quantitative PCR. TIMP-1 (an ADAMTS inhibitor) and TIMP-1 combined with the sIL-13Rα2-Fc intervention increased the levels of GAG and HA, inhibited the expression of type I collagen, and promoted the expression of type II collagen. Conclusion. Adamts8 may participate in the degradation of ECM components such as GAG and HA and lead to an imbalance in the ECM of the intervertebral disc, resulting in intervertebral disc degeneration. sIL-13Rα2-Fc promoted anabolism of the ECM and increased the levels of ECM components by inhibiting the expression of Adamts8, thus maintaining the dynamic equilibrium of the ECM and ultimately delaying intervertebral disc degeneration.

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