富血小板血浆联合主动运动对大鼠髌股关节全层软骨缺损的修复效果研究

The effect of platelet-rich plasma plus active-motion on repair of full-thickness cartilage defects at the patellofemoral joint in rats

目的 探讨富血小板血浆（PRP）联合主动运动对大鼠髌股关节全层软骨缺损修复的效果.方法 SPF级SD大鼠30只,15只用来提取富血小板血浆（PRP）,另外15只随机分成5组（n=6）：A组：空白对照组,B组：造模＋自由活动组,C组：造模＋主动运动组,D组：造模＋ PRP＋自由活动组,E组：造模＋ PRP＋主动运动组.10周后进行大体、组织学、免疫组化观察,检测血清中金属蛋白酶组织抑制因子-1 （TIMP-1）、基质金属蛋白酶-3（MMP-3）含量,计算TIMP-1与MMP-3OD值比值,并进行比较.结果 大体观察、HE染色、免疫组化染色显示：A组,软骨表面光滑,软骨细胞数量多,排列有序,Ⅱ型胶原含量丰富;B组软骨缺损仍较大,表面不平整,成纤维细胞数量最少,Ⅱ型胶原含量最少;C、D、E3组分别进行主动运动或（和）PRP干预后软骨缺损基本修复,表面比较平整,成纤维细胞数量相对较多,Ⅱ型胶原含量相对较多;后3组缺损部位修复的软骨组织基本为纤维软骨组织,很少有透明软骨.各组TIMP-1/MMP-3OD值的比值分别为0.401±0.056、0.247±0.028、0.583±0.054、0.435±0.054、0.387±0.125,差异有统计学意义（F=8.585,P=0.003）.其中B组低于A、C、D、E组,C组高于D、E组,差异均有统计学意义（P＜0.05）.结论 软骨缺损后,早期进行中等强度主动运动或（和）关节腔内注射PRP,对软骨修复有一定的促进作用,尚不能证实主动运动和PRP之间的协同促进作用.

Objective To investigate the effect of platelet-rich plasma（PRP） plus active-motion on the repair of full-thickness cartilage defects at the patellofemoral joint in rats.Methods Thirty SD rats were used for this study.of them,15 were used for preparation of platelet-rich plasma,and the other 15 were randomized into 5 equal groups （n =6）.Group A was subjected to no intervention after sham operation; after modeling of full-thickness cartilage defects at the patellofemoral joint,group B was subjected to free motion,group C to active motion,group D to PRP injection plus free motion,and group E to PRP injection plus active motion.The animals were sacrificed 10 weeks postoperation for gross observation and HE staining to assess the defects,ELISA to assess the serum contents of TIMP-1 and MMP-3,and immunohistochemical staining to detect type Ⅱ collagen.The ratio of TIMP-1 to MMP-3 was calculated and compared between groups.Results Gross observation,HE staining and immunohistochemical staining revealed smooth articular surface and plentiful well-arranged chondrocytes and type Ⅱ collagen in group A,the largest defects with a rough surface and the least type Ⅱ collagen in group B,and defects generally repaired with a relatively smooth surface and more content of type Ⅱ collagen in groups C,D and E.The regenerated tissue was fibrous cartilage rather than hyaline cartilage.The OD value of TIMP-1/MMP-3 was 0.401 ±0.056 in group A,0.247 ±0.028 in group B,0.583 ±0.054 in group C,0.435 ±0.054 in group D,and 0.387 ±0.125 in group E,showing significant differences between groups （F =8.585,P =0.003）.Group B had a significantly lower value than Groups A,C,D and E （P 〈 0.05）.Group C was significantly higher than groups D and E （P 〈 0.05）.Conclusions After articular cartilage defection,early active-motion or （and） PRP injection into the articular cavity may promote repair of cartilage,but the synergistic effect of the two has not been fully confirmed.