The Rabbit Intervertebral Disc Cell Three-diamensions Culture In Vitro And Study On The Effect Of Ozone

Discogenic pain already has become the commen problem to harm the healthy of people. Along with the deep recognition, traditional operation no more is the single choice, and new treatment technics sping up to treat discogenic pain. Chemodiscolysis with ozone with periradicular and periganglionic infiltration is a recent percutaneous techinique widespread in Europe, it has some grace that is security, utility, small damage, rapid recover, etal. Nowaday with rising development of Principles and Practice of the tissue engineering and successfully utilizing to repair bone and articular cartilage defect, we began to try with the tissue engineering to rebuild the disc that had been damaged or removed.Objective:1. Base on the monolayer culture, the study want to establish three diamensions culture model as the experiment groundwork of trasplanting disc tissue to cure discogenic pain.2. This study want to observe the effective of O₂-O₃ mixture in different concentrate at different time through the mixture contributes to intervertebral disc cell cultured in vitro, in order to direct the clinic.Methods:Testâ… : First to set up rabbit intervertebral disc cell monolayer culture, after passage collected the monolayer culture cell to a conical flask, keep giving a eccentric pivot power to revolve(37℃, 160rpm) for six days, after formalin fixation then Bengal gelatine fixiform and embedded in paraffin block to slice for viewing.Testâ…¡: On the other hand, monolayer culture cells were divided to four groups: the first given pure O₂, the others given respective concentration O₃: 30ug/ml, 60ug/ml and 80ug/ml, at 15 and 30 minut obtained cell to make trypan blue test.Statistical analysis: Compared by using statistics software 12.0 to analyze, within grounps using t-test for paired value single, interblock using one-way ANOVA, P＜0.05 consider to have statistical significance. Testâ… finds: Three diamensions culture discovers the cell aggregate look like white sands, i. e. Pallet. After embeding paraffin block and HE dyeing cells gather to take on cell mass, compared with monolayer cell culture showing fusiform shape like mechanocyte, they present round nearly natural.Testâ…¡finds: After 15 minutes different concentration groups all have statistical significance, and along with the concentration of O₃ increasing, the living cells show a lossing tendency. At 30 minutes, statistical result show: besides control group and 30ug/ml group, among others exist statistical significance. At different time, only the 30ug/ml group has statistical significance.Conclude: Dyeing Cell globule cultured through traditionary cell mass aggregation technique in centrifuge tube find that the cells on surface layer is fusiform and internal cells is similar to round or polygon. The phenomenon maybe relate with time length.We discern 30ug/ml O₃ can affect rabbit disc cells, but because the concentration is low, the effective is faultiness; with the extending of the time the effective of 60ug/ml O₃ is all the better flawless; also 80ug/ml O₃ already kill all cells after 15 minutes, cytotoxicity is exorbitance.