Studies of Molecular Changes in Intervertebral Disc Degeneration in Animal Model

Abstract

Objective: To evaluate the structural and molecular changes in the extracellular matrix (ECM) during the process of intervertebral disc degeneration, using animal model. Methods: Wistar rats underwent intervertebral disc degeneration through 20-gauge needle puncture, and 360° rotation applied for 30 sec, representing the degenerated group, while control group was not submitted to this procedure. Histological parameters and expression of extracellular matrix molecules were evaluated in the 15th and 28th days after degenerative induction. Results: Fifteen days after the induction of intervertebral disc degeneration, significant changes were observed, such as reduction in the expression metalloprotease-9 (MMP9) and interleukins (IL-6 and IL-10). There was a significant increase in the expression of vascular endothelial growth factor (VEGF) and caspase-3. However, different alterations in the ECM were observed at 28 days, the level of collagen I, metalloprotease-2 (MMP2) and caspase-3 were enhanced. Furthermore, expression of heparanase isoforms (HPSE1 and HPSE2) mRNA were increased in the degenerative intervertebral disc. Conclusion: The different profiles of ECM molecules observed during the intervertebral disc degeneration suggest that molecular processes such as ECM remodeling, neovascularization, apoptosis and inflammation occur. Experimental Study.

Vascular endothelial growth factor gene variations as a risk predictor in disc degeneration

“…Nonetheless, nucleus pulposus cells could potentially regulate the growth of endothelial cells in a process of disc degeneration through catabolic pathways leading to chronic lumbosciatalgia. ( 26 ) It is not known exactly in which stage of disc degeneration that the neovascularization process is enabled, but some factors, such as smoking, overweight and practice of exercises, can also influence the angiogenesis process. ( 15 )…”

Granulocyte‐colony stimulating factor improves intervertebral disc degeneration in experimental adult male rats: A microscopic and radiological study

“…This incision was performed carefully between the left lateral tail vein and the ventral artery to avoid excessive hemorrhage. After direct access to the targeted tail discs, a sterile 20‐G needle was inserted into the lateral side of the Co5/Co6 and Co7/Co8 discs until the needle reached the nucleus pulposus (NP; Figure 1d,e; de Campos et al, 2016; Li et al, 2016). To ensure a constant depth of needle insertion of 5 mm in all punctured discs, a handmade stopper sleeve was applied to the needle at this distance (Figure 1a; Hua et al, 2016).…”

“…To ensure a constant depth of needle insertion of 5 mm in all punctured discs, a handmade stopper sleeve was applied to the needle at this distance (Figure 1a; Hua et al, 2016). After insertion of the needle, it was rotated 360° and held at its site for 30 s before being withdrawn (de Campos et al, 2016; Li et al, 2016). The Co6/Co7 disc was regarded as a sham‐operated disc as it was exposed without any manipulation.…”

“…An aqueous solution of 3% hydrogen peroxide was used to block the activity of the endogenous peroxidase. The sections were then incubated with anti‐caspase‐3 primary antibodies overnight at 4°C before being incubated with a complex of streptavidin (de Campos et al, 2016).…”

“…Visualizations of SDF‐1α and caspase‐3 expressions were developed by 0.06% 3,3′‐diaminobenzidine staining, and counterstaining was then performed using hematoxylin (de Campos et al, 2016; Ying et al, 2019). A positive expression was indicated by a brown color under light microscope examination.…”