Influence of needle puncture versus abnormal mechanical compression on the biological properties of annulus fibrosus: a preliminary study at nano-scale

doi:10.3969/j.issn.2095-4344.0649

Abstract

Abstract:

BACKGROUND: Currently, the specific pathological mechanism of intervertebral disk degeneration, especially the tissue alteration at nano-scale, is still unknown.

OBJECTIVE: To establish a rat model of intervertebral disk degeneration in the caudal vertebrae and to explore the biomechanical alteration of collagen fibrils of annulus fibrosus at nano-scale induced by needle puncture and abnormal mechanical loading.

METHODS: Eight adult Sprague-Dawley male rats were chosen and the coccygeal vertebral discs from C_5-11 were located under anesthesia. The Co5-Co6 segments were set as normal group. The Co6-Co7 segments were subjected to needle puncture (needle-puncture group). The Co7-Co10 segments were drilled percutaneously by four parallel K-wires and the wires were fixed with self-made braces to make the Co8-Co9 segments under a constant compression environment. The Co8-Co9 segments were set as abnormal compression group. The Co10-Co11 segments were set as adjacent group. Sagittal T2-weighted MRI scanning of the rat caudal vertebrae was performed at 4 weeks after treatment. Hematoxylin-eosin and safranin O/Fast green staining of the intervertebral disc was also performed, and the inner and outer layers of the annulus fibrosus were observed using atomic force microscope.

RESULTS AND CONCLUSION: (1) The MRI signals in the abnormal compression group and needle-puncture group were significantly lower than those in the normal and adjacent groups (P < 0.05), while the MRI signals in normal and adjacent groups showed no significant difference. (2) Histological analysis revealed similarity in the normal and adjacent groups. In the needle-puncture group, the annulus fibrosus on the puncture site arranged disorderly, thickened and bulged inward into the nucleus pulposus, resulting in an unclear boundary between the annulus fibrosus and nucleus pulposus. The number of nucleus pulposus cells on the puncture side was dramatically reduced and vacuole-like structure disappeared. In the abnormal compression group, nucleus pulposus area remarkably decreased, the number of nucleus pulposus cells reduced and vacuole-like structure in the nucleus pulposus disappeared. Boundaries between the nucleus pulposus and annulus fibrosus remained clear. (3) Under the atomic force microscope, the elastic modulus of collagen fibrils in the outer layer was higher than that in the inner layer (P < 0.05). The elastic modulus of collagen fibrils in the adjacent and abnormal compression groups was similar to that in the normal group, while the elastic modulus of collagen fibrils in the needle-puncture group was higher than that in the normal and abnormal compression groups (P < 0.01). To conclude, these results reveal that the needle-puncture can induce remarkable stiffening of collagen fibrils which differs from the abnormal compression.

中国组织工程研究杂志出版内容重点：生物材料；骨生物材料; 口腔生物材料; 纳米材料; 缓释材料; 材料相容性；组织工程

Key words: Intervertebral Disk Degeneration, Microscopy, Atomic Force, Elastic Modulus, Tissue Engineering

CLC Number:

R318

Li Haitao, Liang Ting, Shao Yijie, Chen Xi, Yang Huilin, Luo Zongping. Influence of needle puncture versus abnormal mechanical compression on the biological properties of annulus fibrosus: a preliminary study at nano-scale[J]. Chinese Journal of Tissue Engineering Research, 2018, 22(34): 5526-5517.

Figures/Tables 5

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