Morphology of normal and degenerative nucleus pulposus cells and quantitative analysis of collagen type II protein
10.3969/j.issn.2095-4344.2014.15.012
- VernacularTitle:正常与退变髓核细胞形态学及Ⅱ型胶原蛋白的定量分析
- Author:
Qi WANG
;
Longtao REN
;
Chenggang WEI
;
Junren HE
- Publication Type:Journal Article
- Keywords:
intervertebral disc;
intervertebral disc degeneration;
col agen type II
- From:
Chinese Journal of Tissue Engineering Research
2014;(15):2363-2368
- CountryChina
- Language:Chinese
-
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 col agen type II. OBJECTIVE:To quantitatively observe col agen 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 col ected from adult scoliosis patients and patients with intervertebral disc protrusion, who were al 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 col agen 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 smal amount of fluorescent particles within cells. The expression of col agen type II was decreased significantly compared with normal cells (P<0.05). Safranin O staining showed that, degenerative nucleus pulposus cells began to swel , the nuclei swel ed and were stained slightly, cellprocesses were prolonged, cytoplasmic dyeing was uneven accompanying with vacuole, celldisruption, 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 smal quantity and partial y become apoptotic, the content of col egen type II protein is decreased significantly compared with normal nucleus pulposus cells.
