富血小板血浆联合透明质酸钠对兔膝关节软骨的影响

Efficacy of Platelet-Rich Plasma Combined with Hyaluronic Acid on Rabbit Knee Cartilage

目的:探讨富血小板血浆+透明质酸钠对软骨细胞增殖的影响。方法:48只新西兰大白兔随机分为空白组、富血小板血浆(Platelet Rich Plasma,PRP)组、透明质酸钠(Hyaluronic Acid,HA)组和富血小板血浆联合透明质酸钠(PRP+HA)组,每组12只。PRP组、HA组和PRP+HA组通过改良Hulth法复制兔左膝骨关节炎模型,空白组仅打开膝关节腔后缝合。改良Hulth法6周可建立标准的骨关节炎模型,PRP组和PRP+HA组于造模后第1周取耳静脉血按Aghaloo法制备富血小板血浆,分别将0.5mL PRP和0.5mL PRP+0.5mL HA行左膝关节腔注射,透明质酸钠组0.5mL HA左膝关节腔注射,空白组注射等量生理盐水。1周注射1次,连续注射5周后处死实验动物,取相应标本待检。分别对关节软骨行肉眼下Pelletier评分,苏木精-伊红(HE)染色并进行Mankin评分,Masson染色和甲苯胺蓝染色分别观测胶原纤维及蛋白多糖的变化,透射电镜观察软骨细胞形态变化,ELASA检测关节液Ⅱ型胶原羧基端肽(CTX-Ⅱ)、软骨寡聚基质蛋白(COMP)的表达情况。结果:PRP+HA较单一PRP或HA对兔膝骨性关节炎有更好的治疗效果,可更好地改善Pelletier评分和Mankin评分,差异有统计学意义(P<0.05);减少软骨组织中蛋白多糖及胶原纤维的降解,差异有统计学意义(P<0.05);更好保护软骨细胞微观水平结构;减低关节液中CTX-Ⅱ和COMP的浓度,差异有统计学意义(P<0.05)。结论:富血小板血浆联合透明质酸钠对膝关节骨关节炎软骨病理损伤有一定的保护作用。

Objective:To investigate the efficacy of platelet-rich plasma+hyaluronic acid on the proliferation of chondrocytes.Methods:48New Zealand white rabbits were randomly divided into control group,platelet rich plasma(PRP)group,hyaluronic acid(HA)group and platelet rich plasma combined with sodium hyaluronate(PRP+HA)group with 12rabbits in each group.The left knee osteoarthritis model of rabbit was established by the modified Hulth method in the PRP group,HA group and PRP+HA group.In the control group,the knee joint cavity of rabbit was opened and then sutured only.The standard osteoarthritis model can be established by the modified Hulth method for 6weeks.In the PRP and PRP+HA group,the ear vein blood was collected in the first week after modeling to prepare platelet-rich plasma according to the Aghaloo method.0.5mL PRP and 0.5mL PRP+HA was injected into the left knee joint cavity of rabbit respectively;and the control group was injected with the same volume of normal saline.The animals were injected once a week,and the experimental animals were sacrificed after continuous injection for 5weeks,and the corresponding specimens were taken for examination.The articular cartilage was subjected to Pelletier scores under the naked eye,HE staining and Mankin scores,Masson staining and toluidine blue staining were used to observe the changes of collagen fibers and proteoglycans,and the morphological changes of chondrocytes was observed by transmission electron microscope.The expression levels of CTX-Ⅱand COMP in articular fluid were detected by ELASA.Results:Compared with single PRP or HA,PRP+HA had a better therapeutic efficacy on rabbit knee osteoarthritis,and could improve Pelletier scores and Mankin scores(P<0.05);reduce the proteoglycan and collagen fibers in cartilage tissue(P<0.05);better protection of the microscopic level structure of chondrocytes;the concentrations of CTX-Ⅱand COMP in articular fluid were decreased(P<0.05).Conclusion:Platelet-rich plasma combined with hyaluronic acid has a certain protective efficacy on cartilage pathological damage in knee osteoarthritis.