The Change Of The Nucleus Pulposus When Rat Disc As A Whole Organ System Is Cultured

Objective:Disc degeneration is an important reason for chronic low back pain, how to study its pathogenesis in vitro is currently a hot research.To find a practical method of culturing discs organ system by observing the changes of the nucleus pulposus and the main components of the nucleus extracellular matrix mRNA expression after the whole intervertebral discs, including cartilage eng-plate nucleus,pulposus and anulus fibrosus, were cultivated and evaluating the results of their culture;Methods:400 intervertebral discs were taken out completely from 90 SD rats and put to the culture plate after being divided into 4 groups randomly. Cultivate the whole intervertebral discs with high osmotic culture medium.Osmotic pressure is 410mOsm/kg, then harvest materials at the 0,3,7,14 day and as a standard of different groups and using DAPI double staining with NBT to observe the cell viability, Using HE, Safranin O staining to observe tissue integrity and usingâ…¡collagen immunohistochemical staining andâ… ,â…¡collagen, decorin, aggrecan mRNA expression to observe morphology of them.the comparison was performed between each two groups and the disc tissue culture changes were Observd with belonging of time;Result:The cell viability, tissue integrity and morphology of the whole intervertebral discs were not changed statistically. This shows that:Through our training methods can be successfully cultured in vitro disc complete organs, and can guarantee that within two weeks, no significant changes;Conclusion:We refer to Haschtmann experimental methods to establish the overall culture model of rat disc method Our system could be used to cultivate the whole intervertebral discs for next investigations.