自体富血小板血浆联合骨髓基质干细胞复合改建脱细胞真皮基质修复兔关节软骨的可行性研究
The feasibility study of acellular dermal matrix reconstructed with autologous platelet-rich plasma combined with bone marrow stem cells in the repair of cartilage of rabbit
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摘要: 目的:研究自体富血小板血浆(PRP)联合骨髓基质干细胞(BMSCs)复合改建脱细胞真皮基质(ADM)修复兔关节软骨的修复效果及临床价值。方法:选择60只股骨髁关节损伤的青紫蓝兔作为研究对象,随机分为研究组、对照组和空白组,各20只。首先制备PRP,再分离BMSCs,并进行体外培养,接种于ADM支架上。研究组培养基添加10% PRP,其他条件同对照组,空白组兔不做任何处理。将BMSCs-ADM复合物植入兔关节缺损处,在1、2、3个月末对构建的软骨组织进行免疫力学、组织学、生物力学检测,比较PRP-BMSCs-ADM联合修复关节缺损的效果。结果:随着培养时间延长,BMSCs的细胞数目显著增多,呈现S形生长曲线,第3天后进入对数期,第9天后进入平台期;从第3天开始,细胞数目OD值研究组均明显高于对照组(P<0.01);随着时间的延长,各组标本软骨样细胞数目增多,软骨陷窝逐渐成熟,软骨表面逐渐光滑,IRCS宏观评分和组织学评分均呈现增长趋势(P<0.01),标本压力弹性模量有上升趋势(P<0.05);同时期研究组标本评分与压力弹性模量均显著高于对照组和空白组(P<0.01);3个月后,研究组软骨缺损修复状况明显优于对照组和空白组。结论:自体PRP联合BMSCs复合改建ADM修复兔关节软骨的修复效果良好,具有可行性。Abstract: Objective: To investigate the effects and clinical value of acellular dermal matrix(ADM) reconstructed with autologous platelet-rich plasma(PRP) combined with bone marrow stem cells(BMSCs) in the repair of cartilage of rabbit.Methods: Sixty rabbits with femoral condylar articular injury were randomly divided into the study group,control group and blank group(20 rabbits each group).The PRP and BMSCs were isolated,cultured,and inoculated with the ADM scaffolds in vitro.The 10% of PRP was added to the culture medium in study group,and the other treatment was the same as the control group.The blank group did not give any process.The BMSCs-ADM composite was implanted into the articular defect of rabbits.The reconstructing cartilage tissue was detected using the immune mechanics,histology and biomechanics at the end of 1,2 and 3 months of implantion.The restoration effects of joints defect with PRP-BMSCs-ADM were analyzed.Results: With the prolongation of the culture time,the number of BMSCs cells increased significantly,the growth curve of cells appeared S-shaped,which arrived at logarithmic phase and plateau phase after 3th day and 9th day,respectively.From the 3th day,the OD value of cell counts in study group was significantly higher than that in control group(P<0.01).With the extension of time,the number of cartilage-like cells in all groups increased,the cartilage space matured gradually,the surface of cartilage smoothed gradually,the macro scoring of IRCS and histological scoring increased significantly(P<0.01),the elastic modulus of the specimens pressure increased(P<0.05).During the same period,the specimen scoring and pressure elastic modulus in study group were significantly higher than those in control group and blank group(P<0.01).After 3 months,the repair of cartilage defect in study group was significantly better than that in control group and blank group(P<0.01).Conclusions: The effects of ADM reconstructed with PRP combined with BMSCs in repairing the cartilage of rabbit are good,which is feasible.
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[1] 时长江.PRP联合BMSCs负载改建脱细胞真皮基质对兔膝关节软骨损伤修复作用的研究[D].天津:天津医科大学,2014. [2] 蒋建召.富血小板血浆复合自体浓缩红骨髓促进骨缺损愈合的实验研究[D].郑州:郑州大学,2010. [3] 宋广明.补肾活血汤结合骨髓基质干细胞复合体修复兔膝关节软骨缺损[D].北京:北京中医药大学,2011. [4] 李小建.脱细胞真皮复合骨髓基质干细胞构建组织工程前交叉韧带的实验研究[D].西安:第四军医大学,2013. [5] 李春亮,李文方.膝关节半月板损伤磁共振成像诊断与关节镜检查对比研究[J].临床和实验医学杂志,2013,12(9):687. [6] 贾俊杰.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[D].广州:南方医科大学,2014. [7] SHERIDAN MH,SHEA LD,PETERS MC,et al.Bioabsorbable polymer scaffold for tissue engineering capable of sustained growth factor delivery[J].J Controlled Release,2000,64(1/3):91. [8] 李会波.生物反应器内应用富血小板血浆构建组织工程骨体内血管化的评估的实验研究[D].济南:山东大学,2013. [9] 严安.富血小板血浆凝胶复合材料修复颈椎缺损的实验研究[D].长沙:中南大学,2009. [10] 张寅权,李国东,张平,等.关节镜下FasT-Fix半月板内缝合技术治疗膝半月板损伤[J].临床和实验医学杂志,2012,11(7):507. [11] 李明,张长青.富血小板血浆与关节软骨修复[J].国际骨科学杂志,2010,48(3):171. [12] 李春根,康晓乐,宋广明,等.独活寄生汤结合骨髓基质干细胞复合体修复兔膝关节软骨缺损[J].中国组织工程研究与临床康复,2010,14(32):5957. [13] 陈冬,姚建华,黄炎,等.改良Brostrom法联合踝关节镜治疗踝关节扭伤致慢性踝关节不稳的研究[J].临床和实验医学杂志,2015,10(6):497. [14] PETER SJ,KIM DJ,STAMATAS GN.Effect of transforming growth factor β1 released from biodegradable polymer microparticles on marrow stroma osteoblasts cultured on poly(propylene fumarat)subsrates[J].J Biomed Mater Res,2000,50:452. [15] OKUDA K,KAWASE T,MOMOSE M,et al.Platelet-rich plasma contains high levels of platelet-derived growth factor and transforming growth factor and modulates the proliferation of periodontally related cells in vitro[J].J Periodontol,2003,74(6):849. [16] 徐静,王黎明,周立冬,等.富血小板血浆及脐带间充质干细胞修复软骨损伤[J].中国组织工程研究,2014,18(41):6633. [17] GRUBER R,VARGA F,FISCHER MB,et al.Platelets stimulate proliferation of bone cells:involvement of platelet-derived growth factor,microparticles and membranes[J].Clin Oral Implan Res,2002,13(5):529. [18] 林静,赵景新.大鼠高强度运动后的软骨损伤及寡聚基质蛋白的表达[J].临床和实验医学杂志,2014,09(22):1840. [19] 陈永锋.复合骨髓基质干细胞的β-磷酸三钙在兔脊柱后外侧融合中的应用研究[D].西安:第四军医大学,2012. [20] 张苍宇.组织工程骨膜的构建及其同种异体体内成骨修复兔肩胛骨缺损的实验研究[D].兰州:兰州大学,2014.
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