The Research Of The Activating Effect Of Platelet Rich Plasma (PRP) In Nucleus Pulposus Cells (NPCS) During The Early Intervertebral Disc Degeneration

Objective:To study relevance of the activity as well as proliferation ability of NPCs and IDD,in order to infer the mechanism of IDD.And using PRP to interact with the NPCs during early IDD,to detect whether the PRP could enhance the activity of NPCs so as to promote the repairment of IDD.Methods:1.Use three-month-old NZW rabbits to build the IDD model by needle punching the intervertebral disc.Before the operation,ensure the intervertebral disc is in health by MRI.2.After 2,4and8 weeks,check whether the modeling is successful.Set the normal rabbits as group A,2-week model as group B,4-week model as group C,8-week model as group D.Isolate and culture the NPCs of all the groups until the 2nd generation to test activity of the NPCs.Then detect the related gene by Real-time PCR.3.Extract PRP by Landesberg method using blood of ear artery.4.Use the NPCs of group B interfered by PRP.Simple culture group was set to the control group.Test activity of the NPCs and detect the related gene by CCK-8 and Real-time PCR after 3,4,5,6and 7 days.Results:1.The IDD models were built successfully and the MRI showed that the severity aggravated over time.2.The NPCs showed spindle like shape under LM.After stained by toluidine blue,the cell body was blue,and there were red particles in cytoplasm and around the cell.Immunohistochemistry showed brown granules full of cytoplasm,which matched the characterize of NPCs.3.The activity and soft ossification ability of the NPCs of the 4 groups by CCK-8 and Real-time PCR was A>B>C>D,which decreased over time.4.The PRP was extracted by Landesberg method successfully.The concentration was(1183±13.21)×109/L.5.At the each time point of be intervention with PRP,the activity of NPCs increased,compared with the control group.The activity of NPCs increased not related to the time.Conclusion:1.The IDD model could be built successfully by needle punching the intervertebral disc,and the severity aggravated over time.2.The NPCs could be Isolated by using the pancreatin and be identified by toluidine blue staining and immunohistochemistry.3.The activity and proliferation ability of the NPCs decreased by the develop of IDD.4.After intervention with PRP,the activity and proliferation ability of the NPCs increased,but not related to the time.