滑膜间充质干细胞联合富血小板血浆在软骨损伤修复中的研究
Study of synovial mesenchymal stem cells combined with PRP in cartilage repair
徐训安 1徐同光 1曹鹏飞 1黄永丰 1王强 1何毅 1刘勇1
作者信息
- 1. 苏州高新区人民医院骨科 苏州 215010
- 折叠
摘要
目的:对比滑膜间充质干细胞(SMSCs)、富血小板血浆(PRP)及两者联合的膝关节腔注射3种治疗方式对于兔软骨损伤的修复效果.方法:选取新西兰大耳兔24只,采用手术在兔膝关节中建造软骨损伤模型,采用随机数表法将软骨损伤模型分为空白组、单独SMSCs关节腔注射组(SMSCs组)、单独PRP关节腔注射组(PRP组)和SMSCs联合PRP关节腔注射组(SMSCs+PRP组)4组,每组6只.剔取4组兔关节滑膜进行SMSCs的体外培养,并进行SMSCs的形态学观察及鉴定.抽取兔静脉血,离心制备PRP,对比PRP与全血中血小板及生长因子的含量.将SMSCs和PRP注射入已造模的3组兔膝关节腔中,于注射治疗2、4和6周后根据国际软骨修复协会(ICRS)软骨修复评分对各组的软骨缺损区的修复情况进行评价.结果:兔膝关节滑膜的原代滑膜细胞分离初为圆形,传代后均为梭形.4组的SMSCs表面抗原CD73、CD90和CD105检测阳性率分别为100%、99.22%和99.99%,CD45检测0.5%,具备干细胞的性质.4组的血小板计数显示PRP中血小板浓度约为全血的6倍.PRP中血小板源性生长因子(PDGF)、转化生长因子-β(TGF-β)和血管内皮生长因子(VEGF)的浓度水平分别为(569.15±57.48)ng/ml、(633.56±63.90)ng/ml和(1243.55±106.04)ng/ml,约为全血中的5倍、6倍、7倍.关节腔注射治疗后2周时,4组的软骨修复评分差异均无统计学意义(P＞0.05);注射治疗4周、6周后,SMSCs+PRP组软骨修复宏观形态学及组织学评分较空白组显著提高,差异均有统计学意义(t4周=6.35、9.15,t6周=8.16、8.60,P＜0.05);结论:SMSCs联合PRP对于兔软骨损伤的修复效果明显优于单独进行PRP和单独SMSCs的治疗,且均优于未进行治疗情况,SMSCs联合PRP能够显著提高软骨损伤自我修复的效果.
Abstract
Objective:To compare the repair effects of three kinds of treatment methods included synovial mesenchymal stem cells(SMSCs),platelet rich plasma(PRP)and the combination of them with knee joint cavity injection on cartilage injury of rabbit.Methods:A total of 24 New Zealand rabbits were selected to establish a cartilage injury model of knee joint by using surgery in knee joint of them.The rabbits with cartilage injury model were divided into four groups using a random number table method,which included blank group,single SMSCs with joint cavity injection group(SMSCs group),PRP with joint cavity injection group(PRP group)and the combination of SMSCs and PRP with joint cavity injection group(SMSCs+PRP group),with 6 rabbits in each group.The synovium of four groups of rabbits were scraped off their joints to conduct in vitro culture of SMSCs,as well as the morphological observation and identification of SMSCs.The venous bloods of rabbits were extracted to prepare PRP by centrifugation.The contents of PRP,platelet and growth factor in blood were compared.The SMSCs and PRP were injected into the knee joint cavity of three groups of rabbits with model.After 2,4 and 6 weeks of injection treatment,the repair statuses of cartilages at defection area of different groups were evaluated according to cartilage repair scoring table of International Association for Cartilage Repair(ICRS).Results:The primary synovial cells of rabbit knee joint synovium were initially round in shape after isolation,but almost all of them were spindle shaped after passage.The positive detection rates of SMSCs surface antigen CD73,CD90 and CD105 of 4 group were respectively 100%,99.22%and 99.99%.The CD45 detection was 0.5%,which indicated that they possessed the property of stem cell.The platelet count of 4 groups showed that the platelet concentration in PRP was approximate 6 times of that in whole blood.The concentrations of platelet derived growth factor(PDGF),transforming growth factor-β(TGF-β)and vascular endothelial growth factor(VEGF)were respectively(569.15±57.48)ng/mL,(633.56±63.90)ng/mL and(1 243.55±106.04)ng/mL in PRP,which were approximately 5 times,6 times and 7 times of that in whole blood,respectively.After 2 weeks of injection treatment for joint cavity,there was no significant statistical difference in the scores of cartilage repair among 4 groups(P>0.05).At 4 and 6 weeks of injection treatment,the morphological and histological score of cartilage repair of the SMSCs+PRP group were significantly higher than those of the blank group,and the differences were significant(t4 week=6.35,9.15,t6 week=8.16,8.60,P<0.05),respectively.Conclusion:The repair effect of SMSCs combined with PRP on cartilage injury of rabbit is significantly better than that of single PRP and single SMSCs,respectively,and all of them are better than those without treatment.SMSCs combined with PRP can significantly improve the effect of self-repair of cartilage injury.
关键词
富血小板血浆(PRP)/滑膜间充质干细胞(SMSCs)/软骨损伤/修复Key words
Platelet-rich plasma(PRP)/Synovial mesenchymal stem cells(SMSCs)/Cartilage injury/Repair引用本文复制引用
基金项目
苏州高新区医疗卫生科技计划(2018Q002)
出版年
2024
国家科技期刊平台
NSTL
万方数据