| Background: From the human good history of medicine record, the neck shoulder pain, the pain in waist and lower extremities are contents which its involves, but the intervertebral disc draws back changes also is the neck lumbago most common reason. In 1934, Mixter and Barr first through surgery confirmation and cure waist intervertebral disc prominent oppression nerve root result sciatica, thus founded so-called"the intervertebral disc time (Dynasty of the disc)".The nucleus pulpous excised this classical technique type 70 remaining years of life to cure the innumerable intervertebral disc pathological change patient, however this 70 remaining years of life came, the world every year had several thousand related literature to publish, each time the related orthopedics conference, the intervertebral disc pathological change was still one of most main subjects. This fact also showed that, at present the intervertebral disc drew back the denatured disease the treatment still to have many problems, needed to continue thoroughly the research and the exploration. In the last few years, the organization engineering research also expands to the spinal column surgical department's domain; the overseas scholar has done the organization project intervertebral disc correlation basic research work. Although publishes memoir quantity still were few, however actually responded this research encouraging prospect.Objective: The research and the intervertebral disc denaturalization or the regeneration related environmental factor as well as the intervertebral disc organization and the cell biology behavior, the observation blood platelet derivation growth factor to in vitro raise intervertebral disc cell division multiplication and the matrix synthesis influence, from the cell level, the molecular biology and the gene level carries on the gene treatment and the organization engineering research to the denatured intervertebral disc.Method: 1st, separates and digests the big mouse intervertebral disc organization to obtain the single cell and to complete the original generation of cell vaccination raise.2nd, establishes the control group and the experimental group after the original generation of cytomixis transfer of generation (infliction 1μg/L, 10μg/L, 100μg/L, 1000μg/L different density blood platelet derivation growth factor stimulation growth).3rd, uses HE, the toluidine blue dyeing legal time observes the cellular form, the multiplication change under the inverted micro- scope, the immune cell chemistry dyeing examinationⅡCollagen expression situation.4th, uses MTT (sai zuo blue compared to color) the law determination intervertebral disc cell extinction value and draws up the cell growth curve.5th, will transfer generation the cell digestion for single, after will flow the type cell instrumental measurement to be in the S time cell content surely.6th, draws the conclusion after the statistical software analysis.Results: 1st, observes continuously under the inversion phase contrast microscope, 10 days latter 95% above cell paste the wall, original generation of cell many for spindle-shaped, has the pseudo pod to stretch out; After the pancreas enzyme digestion transfer of generation, the cell pastes the wall growth speed to speed up, but joins the blood platelet derivation growth factor system intervertebral disc cell convergence time to reduce, growth acceleration.2nd, HE dyeing cell many for spindle-shaped, has the pseudo pod to stretch out, the cell nucleus is the circular or the ellipse, the encompassment cell nucleus, in the cytoblastema obviously many vacuoles; The toluidine blue dyeing cell nucleus is the purple, the cytoblastema dyes for the dark blue.ⅡThe collagen immune cell chemistry dyeing presents the masculine result.3rd, MTT the law determination intervertebral disc cell extinction (OD) value may know after the variance analysis examination, the F value is 20.276, P=0.000, explained five group of between comparison differences have the significance. Five groups of between 22 comparisons obtains the control group, 1μg/L the group and 10μg/L between the group compares does not have the difference, 1000μg/L the group and 100μg/L between the group compares does not have the difference, the control group, 1μg/L the group, 10μg/L the group separately with 1000μg/L the group, 100μg/L the group comparison difference has statistics significance, statistics difference obviously (P< 0.05).4th, along with the growth factor density increase, after flows the type cell meter examination to be in the S time cell quantity to present is being connected. rR-PDGF-AA can enhance the cell multiplication activeness, and assumes the dosage effect relations in the effective concentration scope.Conclusion: This experiment provided simple, the effective method for the intervertebral disc cell raise, the blood platelet derivation growth factor multiplied, in the matrix to in vitro raise intervertebral disc cell fission the collagen and the protein polysaccharide synthesis has the promoter action, might enhance the cell line plastochondria activeness obviously, increase the S time cell percentage, its effect in certain scope and dosage and time present correlation. |
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