Abstract:

Objective   To investigate the effects of nucleus pulposus cells apoptosis on degenerated intervertebral discs by using cell amplification with microcarrier system and studying cell apoptosis. Methods   After primary culture of the degenerative nucleus pulposus tissues by operation, nucleus pulposus cells were randomly separated into two groups. Of the two groups, one was cultured with monolayer cluture, another with microcarrier system. In order to detect apoptosis of different stages, Annexin-V-PI staining was used for early stage apoptosis, DeadEndTM staining for late stage apoptosis, immunohistochemical staining for Bcl2 protein. Results   Annexin-V-PI staining demonstrated that apoptosis rate of nucleus pulposus cultured with microcarrier system was lower in stable phase (P<0.05). However, there was no significant difference between the two different culture methods in logarithmic growth phase (P>0.05).  The data of DeadEndTM staining and immunohistochemical staining showed that apoptosis rate of nucleus pulposus cultured with microcarrier system was lower in both phases (P<0.05). Conclusion   Microcarrier system, which can efficiently reduce the apoptosis rate of nucleus pulposus cells, is a promising culture method applied in tissue engineering of intervertebral discs.

Key words: Microcarrier; Apoptosis; Cell culture; Neucleus pulposus; Degenerative