Morphology of normal and degenerative nucleus pulposus cells and quantitative analysis of collagen type II protein

doi:10.3969/j.issn.2095-4344.2014.15.012

Abstract

Abstract:

BACKGROUND: The narrowing of intervertebral space induced by the intervertebral disc degeneration is mainly characterized by the expression of proteoglycan in nucleus pulposus cells and the reduction of collagen type II.
OBJECTIVE: To quantitatively observe collagen type II protein in adult normal and degenerative intervertebral disc nucleus pulposus cells by immunofluorescence staining and safranin O staining.
METHODS: The nucleus pulposus specimens were collected from adult scoliosis patients and patients with intervertebral disc protrusion, who were all volunteers. After culture, 26 cells in each patient were measured. There were 78 cells in both normal group and degeneration group. The normal and degenerative intervertebral disc nucleus pulposus cells were subjected to safranin “O” staining, and gray values were determined; intracellular collagen type II was detected by immunofluorescence staining.
RESULTS AND CONCLUSION: Immunofluorescence staining revealed that, degenerative intervertebral disc nucleus pulposus cells were only mildly stained, with the fuzzy staining, the shape was round, spindle, fusiform and irregular. There were a very small amount of fluorescent particles within cells. The expression of collagen type II was decreased significantly compared with normal cells (P < 0.05). Safranin O staining showed that, degenerative nucleus pulposus cells began to swell, the nuclei swelled and were stained slightly, cell processes were prolonged, cytoplasmic dyeing was uneven accompanying with vacuole, cell disruption, scattered and chaotic distribution were visible, patches of necrosis were observed. The image gray value showed no significant difference compared with normal nucleus pulposus cells (P > 0.05). The degenerative intervertebral disc nucleus pulposus cells have a small quantity and partially become apoptotic, the content of collegen type II protein is decreased significantly compared with normal nucleus pulposus cells.

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

全文链接：

Key words: intervertebral disc, intervertebral disc degeneration, collagen type II

CLC Number:

R318

Wang Qi, Ren Long-tao, Wei Cheng-gang, He Jun-ren. Morphology of normal and degenerative nucleus pulposus cells and quantitative analysis of collagen type II protein[J]. Chinese Journal of Tissue Engineering Research, 2014, 18(15): 2363-2368.

Figures/Tables 2

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