摘要
背景:椎间盘器官体外培养模型作为体内试验和细胞培养的联系,能够在原生基质中维持细胞活性,从而保留了重要的细胞与细胞及细胞与基质的相互作用。目的:观察体外培养兔脊柱运动节段模型髓核组织的变化。方法:将13只新西兰白兔处死后在无菌条件下取出脊柱运动节段50个,在高渗培养基中进行整体培养(410 m Osm/kg),于培养前及培养后第3,7,14,21天,各取10个椎间盘分别进行苏木精-伊红染色、Ⅱ型胶原免疫组织化学、蛋白多糖含量和髓核细胞活力测定。结果与结论:培养14 d苏木精-伊红显示椎间盘组织结构基本保持完整,21 d椎间盘组织形态学破坏;Ⅱ型胶原免疫组织化学染色强度14 d无显著差异(P>0.05),21 d染色变浅,与之前各时间点相比有显著差异(P<0.05);蛋白多糖PAS/AB染色7 d内强度无降低,14 d强度有所减弱,21 d染色强度进一步减弱;髓核细胞荧光检测21 d髓核细胞荧光强度减弱,细胞活性降低,与之前各时间点比较差异明显(P<0.05)。结果表明,培养14 d兔脊柱运动节段椎间盘髓核组织退变不显著,培养21 d模型髓核退变明显,脊柱运动节段目前在14 d内可作为研究生物力学对椎间盘影响的体外实验模型。
BACKGROUND： In vitro organ culture model of the intervertebral disc as a contact between in vivo tests and cell culture can maintain cell activity in a native matrix, thus retaining the important cell-cell and cell-matrix interactions. OBJECTIVE： To observe the changes in the nucleus pulposus during in vitro culture of rabbit spinal motion segments. METHODS： A total of 50 spinal motion segments were harvested from 13 New Zealand white rabbits under aseptic conditions. These specimens were maintained for organ culture with hyperosmotic media（410 m Osm/kg）, then 10 discs were observed by hematoxy-eosin staining and immunohistochemistry of type II collagen as well as proteoglycan content and viability of nucleus pulposus cells were determined before and at 3, 7, 14, 21 days after culture, respectively. RESULTS AND CONCLUSION： Hematoxylin-eosin staining showed the structure of intervertebral disc tissue remained intact after 14 days of culture, but severely degenerated at 21 days of culture. The intensity value of type II collagen immunohistochemical staining in the nucleus pulposus had no significant changes within 14 days（P 〉 0.05）, but the staining became shallow at 21 days, which was significantly different from that at 3, 7, 14 days（P 〈 0.05）. PAS/AB staining of proteoglycan of the nucleus pulposus showed no decrease in tinting strength within 7 days, but the strength became weakened slightly at 14 days and further weakened at 21 days. The intensity value of fluorescence staining of nucleus pulposus cells decreased at 21 days and the viability of nucleus pulposus cells decreased significantly as compared with that at 3, 7, 14 days（P 〈 0. 05）. These findings indicate that the degeneration of the nucleus pulposus of rabbit spinal motion segment showed no significant changes at 14 days of culture, but became remarkable at 21 days of culture, indicating that the spinal motion segement can be used as an in vitro experimental model to study the biomechanical changes of the intervertebral disc within 14 days of culture.
出处
《中国组织工程研究》
CAS
北大核心
2015年第51期8241-8246,共6页
Chinese Journal of Tissue Engineering Research
基金
骨关节退行性病变中医防治创新团队(YS1304)
骨与关节退行性病变研究专项课题(WJYY2014-ZX-06)~~
