自体富血小板血浆纳米球凝胶缓释系统对兔膝关节软骨损伤的修复效果研究  

Study of autologous platelet-rich plasma loaded nanoparticle hydrogel sustained release system on rabbit kneel articular cartilage damage repair

作　　者：缪小兵[1] 周友浪 居飞 谭军 MIAO Xiaobing;ZHOU Youlang;JU Fei;TAN Jun(Department of Orthopedics,the People’s Hospital of Rugao,Jiangsu 226500;Department of Hand Surgery,the Affiliated Hospital of Nantong University)

机构地区：[1]如皋市人民医院骨科,江苏226500 [2]南通大学附属医院手外科

出　　处：《交通医学》2021年第1期5-9,F0002,共6页Medical Journal of Communications

基　　金：国家自然科学基金(81871824,81672231,81871763)。

摘　　要：目的:建立负载自体富血小板血浆纳米球凝胶缓释系统,观察对兔膝关节软骨损伤的修复效果。方法:建立兔双膝关节股骨内侧髁软骨面缺损模型,随机分成空白组、凝胶组和纳米球凝胶组各9只。兔心脏采血采用二次离心法制备自体富血小板血浆(PRP),将PRP加入透明质酸水凝胶中制成PRP凝胶。将PRP先负载到乳酸-羟基乙酸共聚物(PLGA)纳米球中,然后再加入透明质酸水凝胶中,建立PRP纳米球凝胶缓释系统。分别将PRP凝胶、PRP纳米球凝胶缓释系统应用于兔膝关节软骨缺损面。术后4周、8周、12周处死实验动物,对兔膝关节标本进行大体观察,采用国际软骨修复协会(ICRS)宏观修复评价量表进行评价。对损伤区进行组织取材,切片后行HE染色和番红固绿染色,应用O’Driscoll评分标准进行组织学评分。结果:凝胶组在4周、8周、12周时ICRS宏观修复评分分别为5.67±0.52分、8.83±0.75分和9.00±0.63分,纳米球凝胶组分别为5.67±0.52分、9.50±0.55分和10.16±0.75分,均高于空白组的2.50±0.58分、4.00±0.82分和4.25±0.50分,差异均有统计学意义(P<0.05),凝胶组、纳米球凝胶组8周、12周ICRS宏观修复评分高于4周,差异均有统计学意义(P<0.05),但凝胶组与纳米球凝胶组的差异无统计学意义(P>0.05)。空白组在4周、8周、12周时O’Driscoll组织学评分分别为4.00±0.82分、4.25±0.96分和4.00±0.82分,凝胶组分别为9.33±1.03分、11.50±1.05分和10.00±1.79分,纳米球凝胶组分别为10.83±1.17分、15.00±1.41分和13.50±1.87分,三组间差异均有统计学意义(P<0.05),纳米球凝胶组评分最高。结论:自体富血小板血浆凝胶和自体富血小板血浆纳米球凝胶缓释系统均具有软骨修复作用,可以促进局部软骨基质形成和软骨细胞聚集,但超过8周时软骨修复作用减退。Objective:To establish an autologous platelet-rich plasma loaded nanoparticle hydrogel sustained release system and observe its therapeutic effect on articular cartilage injury in rabbits.Methods:Cartilage defect models were manufactured in the rabbit femur medial condyle.They were randomly divided into three groups,9 in blank group,9 in gel group and 9 in nanoparticle gel group.Platelet rich plasma(PRP)was obtained from rabbit heart blood by two centrifugation.PRP was added into hyaluronic acid hydrogel to prepare PRP gel.PRP was first loaded into PLGA nanoparticle and then added into hyaluronic acid hydrogel to establish PRP nanoparticle hydrogel sustained release system.PRP gel and PRP nanoparticle hydrogel sustained release system were applied to the cartilage defect surface of rabbit knee respectively in the gel group and the nanoparticle gel group.The blank group was not treated.The experimental animals were killed at 4,8 and 12 weeks after operation.Firstly,the rabbit knee specimens were observed in general and evaluated by Interna tional Cartilage Repair Society macroscopic scoring scale.Then the tissue samples were taken from the injured area,and the sections were stained with hematoxylin eosin and safranin o-fast green.The O’Driscoll scoring standard was used for histological scoring.Results:At 4 weeks,8 weeks and 12 weeks,the macroscopic repair scores of the gel group were 5.67±0.52,8.83±0.75 and 9.00±0.63 respectively.Those of the nanoparticle gel group were 5.67±0.52,9.50±0.55 and 10.16±0.75 respectively,which were higher than those of the blank group 2.50±0.58,4.00±0.82 and 4.25±0.50.The difference was statistically significant(P<0.05).The macroscopic repair scores of the gel group and the nanoparticle gel group at 8 weeks and 12 weeks were all higher than at 4 weeks,the difference was statistically significant(P<0.05),but the difference between gel group and the nanoparticle gel group was not statistically significant(P>0.05).At 4 weeks,8 weeks and 12 weeks,O'Driscoll histological scores

关 键 词：富血小板血浆 纳米球凝胶缓释系统 软骨损伤 兔 

分 类 号：R336[医药卫生—人体生理学]