| Part Ⅰ Therapeutic effect of local application of pure platelet-rich plasma on intervertebral disc degeneration in ratsObjective:Investigate the therapeutic effect of local application of pure platelet-rich plasma(P-PRP)on intervertebral disc degeneration in rats.Method:20 rats were randomly divided into experimental group(n=15)and control group(n=5).Rats in the experimental group used annulus puncture to construct a model of intervertebral disc degeneration.After confirming the success of the model by MRI examination,5 rats in the experimental group were randomly selected,and the disc samples were taken for chemical staining after being killed.The rats in the control group were also killed and the samples were taken for chemical staining.The blood of the remaining rats in the experimental group was collected to prepare P-PRP.Rats in the experimental group were injected with about 5u L autologous P-PRP(P-PRP group)into the intervertebral disc of Co7/8,and the same amount of phosphate buffer saline(PBS)(PBS group)was injected into the intervertebral disc of Co6/7.MRI examinations were performed at 2,4,and 8 weeks after injection,and disc degeneration was graded by Pfirrmann grading method to quantify the signal intensity of the intervertebral disc.After the last MRI examination,the rats were killed to obtain the disc samples,and the morphology of the intervertebral discs and the concentration of extracellular mechanism were observed and estimated by HE,Safranine O-Fast green staining,Masson staining and immunohistochemical staining.The degree of intervertebral disc degeneration was evaluated by Nishimura histological score.Result:MRI confirmed that the models were successful,and there was no significant difference in the grades of disc degeneration between PBS group and P-PRP group(P<0.05).At 2,4 and 8 weeks after injection,the signal of intervertebral disc in PBS group decreased continuously.On the contrary,the signal of T2 image in PRP group gradually improved with time.The grade of disc degeneration in P-PRP group was lower than that in PBS group at each time point.At 4 and 8 weeks after injection,the signal intensity of PRP group was significantly better than that of PBS group(P<0.05),suggesting that P-PRP can make the degenerative disc recover from low signal to high signal.According to the statistical results,the grade of disc degeneration in P-PRP group was significantly lower than that of PBS group at the 8th week after injection,which indicated that P-PRP can recover the low signal of degenerative disc to high signal.HE staining showed that the structure of annulus fibrosus was disordered and nucleus pulposus was obviously shrunk in PBS group;In P-PRP group,the structure of annulus fibrosus was still clear,the arrangement was slightly disordered,and the area of nucleus pulposus was not significantly reduced.According to Nishimura histological score,the score of PRP group was significantly better than that of PBS group(P<0.05).Safranine O Fast green staining showed that the red staining of proteoglycan in nucleus pulposus of PRP group was more uniform and darker than that of PBS group.Masson staining showed that the blue stained collagen in PRP group was significantly darker than that in PBS group,suggesting that the content of collagen fibers in the disc was increased.The results of immunohistochemistry showed that the expression of type II collagen in P-PRP group was significantly higher than that in PBS group.Conclusion:The results of animal experiments show that P-PRP injection can delay the degeneration development,and P-PRP has a certain repair function on the degenerative disc.Part Ⅱ Efficacy of local application of leukocyte depleted platelet rich plasma in the treatment of discogenic low back painObjective:To evaluate the efficacy and safety of P-PRP percutaneous injection in the treatment of discogenic low back pain.Method:The study was approved by medical ethics committee of Affiliated Hospital of Qingdao University.From June 2018,the patients who met the inclusion criteria and exclusion criteria were selected strictly,and the general information of patients was collected.P-PRP was prepared by collecting the patient’s autologous blood before the operation,then PRP was injected into the intervertebral disc of the degenerative segment,and patients were followed up for up to 1 year after the operation.The pain scores,functional scores and MRI manifestations of intervertebral disc were statistically analyzed before and after treatment.Result:Finally,10 eligible patients were included,with an average age of 32.0±8.7 years.The average concentration of platelets in the patient’s whole blood was(255.6±40.8)×10~9/L,the average concentration of platelets in P-PRP was(967.9±124.7)×10~9/L,and the platelet concentration in PRP was approximately 3.78 times that in whole blood.Before treatment,the average VAS and ODI scores were 7.4±0.9 and34.9±4.8,respectively.After 1 week,4 weeks,12 weeks,24 weeks,48 weeks and 1 year after treatment,the patients’VAS and ODI scores were significantly improved(P<0.05,respectively).The improvement was most obvious at 12 weeks after treatment.At this time,the VAS and ODI scores were 3.1±0.9 and 20.1±4.7,respectively.MRI showed that at 1year after the treatment,the signal of the intervertebral disc in the treated segment of the patient was improved compared with that before the treatment.There were no adverse reactions and complications during the whole study.Conclusion:Percutaneous intradiscal injection of P-PRP is a safe and promising new treatment strategy that can effectively relieve discogenic low back pain. |
PDF Full Text Request