富血小板血浆对兔膝关节滑膜炎的治疗作用

EFFECT OF PLATELET-RICH PLASMA ON SYNOVITIS OF RABBIT KNEE

目的富血小板血浆(platelet-rich plasma,PRP)对软组织修复、创伤愈合及骨缺损修复均有促进作用,通过建立蛋白酶诱导的兔膝骨性关节炎模型并以PRP进行干预,探讨PRP对滑膜炎的抑制及治疗作用。方法取6月龄清洁级新西兰大白兔20只,雌雄不限,体重2.5～3.0 kg;于兔耳中央动脉取血采用Landesberg等的方法制备PRP,同时对外周血及PRP进行血小板计数及PDGF、TGF和VEGF浓度检测。于20只兔双侧膝关节内分别注射0.3 mL 4%木瓜蛋白酶建立膝骨性关节炎模型后,随机分为实验组和对照组(n=10)。实验组:双侧膝关节每周注射0.3 mL PRP,共10周;对照组:注射无菌生理盐水。于第1次注射后第2、4、6、8、10周取血检测红细胞沉降率及IL-1β浓度,取膝关节滑膜标本行大体及HE染色观察,并根据Mankin评分标准对滑膜评分。结果血小板计数示PRP中血小板浓度约为外周血的6.8倍。PRP中PDGF、TGF-β和VEGF的浓度分别约为外周血中的5、8、7倍,差异均有统计学意义(P<0.05)。所有实验动物均存活至实验完成。实验组第2、6、8、10周红细胞沉降率与对照组比较,差异均有统计学意义(P<0.05),第4周组间比较差异无统计学意义(P>0.05)。实验组第4、6、8、10周IL-1β浓度与对照组比较,差异均有统计学意义(P<0.05),第2周组间比较差异无统计学意义(P>0.05)。对照组关节滑膜广泛水肿、增厚,表面纤维素性渗出,标本滑膜表面偶见血管翳样组织出现;实验组自第2周开始,可见滑膜渗出减少,随时间延长表面充血现象减轻,水肿程度下降,第10周时可见标本滑膜略增厚,未见明显水肿,表面少量淡黄色关节液覆盖,无粘连现象。实验组第4、6、8、10周Mankin评分与对照组比较,差异均有统计学意义(P<0.05),第2周组间比较差异无统计学意义(P>0.05)。结论 PRP通过修复受损组织、抑制炎性因子释放等途径,对蛋白酶诱导的兔滑膜炎发挥缓解和治疗作用。

Objective Platelet-rich plasma （PRP） can promote the repair of soft tissue, wound, and bone defect. To investigate the effect of PRP on synovitis by establ ishing papain-induced osteoarthritis model of rabbit knee and interfering with PRP. Methods Twenty healthy 6-month-old rabbits （male or female, weighing 2.5-3.5 kg） were randomly divided into the experimental group （n=10） and the control group （n=10）. The whole blood （10 mL） was extracted from the central aural artery and PRP was prepared with the Landesberg’s method. Meanwhile, the platelet derived growth factor （PDGF）, transforming growth factor （TGF）, and vascular endothel ial growth factor （VEGF） concentrations in the circulating blood and PRP were measured. The 4% papain solution （0.3 mL） was injected into the knee joint cavity to establ ish the osteoarthritis model. After that, PRP （0.3 mL） was injected into the knee joints every week for 10 weeks in the experimental group, while normal sal ine of the same volume in the control group. At 2nd, 4th, 6th, 8th, and 10th weeks after the first injection, the erythrocyte sedimentation rate （ESR） and interleukin 1β （IL-1β） concentrations in the whole blood were tested, and the histological changes of the synovium were observed by HE staining and the Mankin scores were made. Results The blood cell counting showed that the platelet concentration of PRP was 6.8 times as that of the circulating blood. PDGF, TGF-β, and VEGF were 5, 8, and 7 times as those of the circulating blood, showing significant differences （P 〈 0.05）. All animals survived to the end of experiment. There were significant differences in the ESR at 2nd, 6th, 8th, and 10th weeks and in the IL-1β at 4th, 6th, 8th, and 10th weeks between 2 groups （P 〈 0.05）. In the control group, the synovium was edematous and thickened with fibrous effusion and pannus on surface; in the experimental group, the effusion of the synovium was decreased and less congestion and edema were observed at the 2nd week; the synovium was observed to be a bit thickened without obvious edema, with sl ight amount of yellowish joint fluid on surface and no conglutination at the 10th weeks. There were significant differences in the Mankin score at 4th, 6th, 8th, and 10th weeks （P 〈 0.05） between 2 groups. Conclusion PRP is beneficial to the alleviation of synovitis induced by papain according to restoring the damaged tissue and depressing the inflammatory factors.