The Feasibility Of Nuclear Pulposus-like Cells Complicated With Alginate Treatment For Degenerated Intervertebral Disc

【Objective】1.To testify the capable of intervertebral disc degeneration model introduced by Annularacupuncture.2.To observe what happened to collage type II、aggrecan、MMP-3and IL-1of thedegenerated intervertebral disc after the transplantation of nuclear pulposus-like cellscomplicated with Alginate.【method】(1)First，rabbit bone derived mesenchymal stem cells (BMSCs) and nucleus pulposuscells were achieved、cultured and expanded，then co-culture model which contains bothBMSCs and nucleus pulposus cells were established in the Transwell chamber.(2)On theother hand L3/4、L4/5、L5/6intervertebral disc degeneration of18rabbit were inducedby Annular acupuncture;(3)once it’s conformed by MRI two weeks later, the BMSCswhich co-cultured with nucleus pulposus cells(NPC) in the Transwell chambercomplicated with alginate and the alginate with no cell were transplanted into two of thedegenerated intervertebral discs randomly(experiment group and conditional experimentgroup); nothing more than annular acupuncture was did to the other one (no treatmentcontrol group).(4)4、8、12weeks later specimen were collected, immunohistochemistryand pro plus were used to compare if there is a different of proteoglycan and collagetype II between each group.and western blotting were used to compare if the contain ofMMP-3and IL-1were reduced.【result】MRI showed that the disc of all the18rabbits degenerated2weeks later afteranu-puncture. BMSCs immunohistochemistry, three pages after coculture, showsstrong positive both in proteoglycan and collage type II;immunohistochemistry foundthat aggrecan and collagen type II in the experiment group,4、8、12weeks later aftersurgery, increased,and is higher than the other two group.as time passed by,aggrecan and type II collagen level in the experiment group augmented,and they were significantlyhigher than that in the blank group and control group (both P<0.05);MMP-3and IL-1inthe control group on the other hand reduced,as time passed by,and were smaller than theother two group.【conclusion】1.Annular acupuncture can successfully conduce disc degeneration of the rabbit2.Nuclear pulposus-like cells complicated with Alginate can not only increase collagentype II and proteoglycan, in the same time，it can also reduce MMP-3and IL-1in thedegenerated disc.