| OBJECTIVE:This study aims to observe the changes of the process of early intervertebral disc degeneration of rabbits.Meanwhile the changes in terms of iconography,cytology and histology after platelet rich plasma treating intervertebral disc degeneration are also observed.METHODS:Sixty healthy New Zealand rabbits are obtained which are about three month old and weigh about 2.0?3.0 kilogram with half male and female.Several rabbits are randomly obtained to get 5?10 milliliter blood from ear vein respectively.2 milliliter blood of each rabbit are preserved for platelet counting.The remaining blood are prepared for platelet rich plasma extraction using the method of Landesberg twice centrifugation method.Platelet counting after extraction is also performed.Sixty rabbits are randomly divided equally in five groups which are blank control group,sham operation group,PBS group,NPC group and PRP group.Only sham operation group receives the operation of exposing intervertebral disc rather than puncturing while blank control group receives nothing.Others receive the degeneration operation on the second and third intervertebral discs.All rabbits receive MRI to explore the degree of degeneration after two weeks and accept intervention operation directly.PBS group:the degenerated intervertebral disc was injected 0.1 milliliter PBS;NPC group:the degenerated intervertebral disc was injected 0.1 milliliter NPC;PRP group:the degenerated intervertebral disc was injected 0.1 milliliter PRP.Four rabbits are randomly selected from each group to perform iconography,histology and molecular biology detection after two,four,eight weeks respectively.The detection of intervertebral disc degeneration contains MRI,HE staining,Masson staining and PCR.RESULTS:1.In microscope,long fusiform and intensively distributed nucleus pulposus was seen in none degeneration groups.Nucleus pulposus cells in degeneration groups grow slow,poorly distributed and varied morphology.2.Masson Staining shows that the collagen in none degeneration intervertebral discs is much more than degeneration intervertebral discs.The collagen in NPC and PRP groups is thicker and larger than PBS group.The collagen density and area of PRP group is bigger than NP group.The collagen is also bigger and more intensive in PRP group at high power.HE Staining shows cells are more and intensively distributed in none degeneration groups.There is no obvious changes in cells and matrix distribution in the degeneration groups after 2 weeks intervention.Matrix change can be seen in the 4th week of intervention while cells decrease in the 8th week.However,there is still a lot of cells can be found in matrix in PRP group in the 8th week of intervention.3.MRI T2 shows that the signal is generally high in none degeneration groups while it is low in degeneration groups.The T2mapping value is higher in none degeneration groups than degeneration groups.The disc height of blank control group is as same as fake operation group.And the difference value of disc height is also same in three degeneration groups.The T2mapping difference value of/nucleus pulposus and platelet rich plasma group is smaller than that of phosphate buffered saline in the second disc after 4 weeks intervention.And it is same between nucleus pulposus and platelet rich plasma group.The same result was found in the third disc after 8 weeks intervention.4.PCR result shows that Col1A1,Col ? and Sox9 have the same trend.The expression of Col1A1 gene goes up in NPC group and PRP group after 2 weeks intervention.This result is meaningful.However,The expression of Col1A1 gene goes up in PRP group only after 4 weeks intervention while NPC group not.The expression of Col ? and Sox9 have the same trend.So PRP and NPC can interve intervertebral disc degeneration in different degree.And PRP can last longer than NPC according to the activity of degenerated nuclus pulposes cells in degenerated intervertebral disc.CONCLUSION:Platelet rich plasma can effectively slow down the process of intervertebral disc degeneration.There is no significant difference of curative effect between platelet rich plasma and nucleus pulposes when treating intervertebral disc degeneration.Both of them have intervetion effect in the process of intervertebral disc degeneration. |
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