不同血小板浓度富血小板血浆对动物软骨细胞增殖的影响及其治疗膝骨关节炎的临床效果观察  

Effect of platelet-rich plasma with different concentrations on chondrocyte proliferation and its clinical effect on knee osteoarthritis

作　　者：许育兵[1] 刘广亚[1] 朱展鸿 张卫[1] 施琳颖[1] 单桂秋[1] XU Yubing;LIU Guangya;ZHU Zhanhong;ZHANG Wei;SHI Linyin;SHAN Guiqiu(Department of Blood Trafusion,Guangzhou General Hospital of Guangzhou Military Command,Guangzhou 510010,China.)

机构地区：[1]广州军区广州总医院输血科

出　　处：《中国输血杂志》2018年第6期583-587,F0001,共6页Chinese Journal of Blood Transfusion

基　　金：军队后勤科研计划项目(BGZ15C002);广州市科技计划项目(201508020115);广东省医学科学技术研究基金(A2018221)

摘　　要：目的探讨富血小板血浆（PRP）中不同血小板浓度（Plt）对软骨细胞增殖和膝骨关节炎治疗效果影响。方法软骨细胞增殖实验选择小鼠软骨细胞做细胞培养：将人浓缩血小板分成4个实验组,分别调整Plt（×109/L）为A组200、B组500、C组1 000、D组1 500,对照组以DMEM/F12培养基加入5%的胎牛血清;采用CCK-8检测法检测不同Plt的PRP对小鼠软骨细胞在24、48、72 h的增殖效果。临床疗效观察选取膝关节骨性关节炎120例作为研究对象,按照上述细胞增殖实验的4种Plt的PRP随机均分为A、B、C、D 4组（n=30）;用EDTA抗凝无菌试管采集受试患者静脉血40 m L/（人）次,使用富浆法离心分离浓缩血小板,调整血小板达到上述4种Plt,治疗4次/例;采用Lysholm膝关节评分量表法评估不同Plt的PRP在治疗前和治疗后1、3、6个月4个时间（点）对骨性关节炎的疗效。结果软骨细胞增殖实验（OD值）：对照组及A、B、C和D 4个实验组在小鼠软骨细胞培养24 h时,分别为0.64±0.01、0.77±0.03、0.84±0.01、0.87±0.02、0.88±0.13（P〈0.05）;48 h时分别为0.86±0.01、1.03±0.10、1.29±0.06、1.32±0.01、1.40±0.04（P〈0.05）;72 h时分别为1.18±0.07、1.58±0.03、1.79±0.08、1.90±0.04、1.92±0.02（P〈0.05）。其中4个实验组组间比较,24、48、72 h时B、C、D 3组均明显优于A组（P〈0.05）,同时C、D 2组又明显优于B组（P〈0.05）。PRP治疗膝关节骨性关节炎疗效（膝关节功能评估以Lysholm评分表示）：与治疗前比较,除A组治疗前后无明显改善（P〉0.05）外、B、C、D 3组治疗后1个月的评分为69.93±2.43、68.57±2.13、66.98±2.33,3个月为70.50±3.22、73.20±2.44、71.24±3.75,6个月为72.10±2.54、75.13±3.18、74.80±2.43（均为P〈0.05）;其中组间比较,C组明显优于B组（P〈0.05）;6个月时,C、D 2组明显优于B组（P〈0.05）。结论 Plt为1 000×109/L的PRP对小鼠软骨细胞增殖效果最为明显,且能获得PRObiective To investigate the effect of platelet-rich plasma（ PRP） with different concentrations on chondrocyte proliferation and its clinical effect on knee osteoarthritis. Methods Chondrocyte proliferation： Blood platelets were collected from healthy subjects and divided into 4 treatment groups and 1 control group. The platelet concentration in the treatment groups are as follows： 200×109/L in group A,500×109/L in group B,1 000×109/L in group C and 1 500×109/L in group D. DMEM/F12 medium with 5% fetal bovine serum was added in the control group. Mouse chondrocyte was selected for cell proliferation experiment. The proliferation effects of PRP with different concentrations on chondrocytes were detected by CCK-8 assay at the time point of 24,48 and 72 h. Clinical effect on knee osteoarthritis： A total of 120 patients with knee osteoarthritis were recruited in this study and randomly divided into 4 groups（ A,B,C,D,n = 30） according to the 4 PRP concentrations mentioned previously. 40 m L of peripheral blood was collected from each patient by EDTA anticoagulant sterile test tube method. Platelets were separated and concentrated by the rich-plasma method and then adjusted to appropriate concentration for knee injection. Each patient were treated with 4 injections. The effects of PRP with different concentrations on knee osteoarthritis were evaluated by Lysholm Knee Joint Score at 4 different time points,before the treatment and 1,3,6 months after the treatment,respectively. Results Results Chondrocyte proliferation was evaluated by the OD value of CCK-8 assay. The OD value in group A,B,C,D and the control was 0. 64±0. 01,0. 77±0. 03,0. 84±0. 01,0. 87±0. 02,0. 88±0. 13 at 24 h（ P〈0. 05）,0. 86 ± 0. 01,1. 03 ± 0. 10,1. 29 ±0. 06,1. 32±0. 01,1. 40±0. 04 at 48 h（ P〈0. 05）,and 1. 18±0. 07,1. 58±0. 03,1. 79±0. 08,1. 90±0. 04,1. 92±0. 02 at 72 h（ P〈0. 05）,respectively. The comparison among the 4 treatment groups showed that cell proliferation in group B,C and D were better t

关 键 词：富血小板血浆 膝骨关节炎 软骨细胞 细胞增殖 PRP浓度动物实验 小鼠 

分 类 号：R331.1[医药卫生—人体生理学] R457.1[医药卫生—基础医学]