脂肪间充质干细胞用于治疗骨性关节炎的临床实验研究

发布时间：2019-03-29 11:52

【摘要】：目的:探讨兔同种异体ADSCs细胞膜片可以作为软骨缺损修复的种子细胞;且在兔膝关节腔内微环境作用下可被诱导生长分化成透明软骨组织。人自体脂肪间充质干细胞(Human adipose derived mesenchymal stem cells,hADSCs)复合玻璃酸钠、富血小板血浆(platelet rich plasma,PRP)注射治疗膝关节骨性关节炎(Osteoarthritis,OA)的安全性和有效性。方法:实验一:(1)取新西兰大白兔腹股沟脂肪组织,分离、培养脂肪间充质干细胞。制备成ADSCs膜片。(2)新西兰大白兔膝关节股骨髁部软骨损伤造模后行ADSCs膜片移植。(3)ADSC修复兔软骨损伤大体照及组织学评价。实验二:(1)取人腹部脂肪组织,分离获取human ADSCs,经体外培养获得足量的ADSCS,并以流式细胞仪检测其表面标志物,分析成软骨细胞分化能力。(2)选择咸阳市第一人民医院骨科中心门诊30例膝关节轻中度骨性关节炎患者,关节腔内注射人ADSCs复合玻璃酸钠、富血小板血浆(PRP),评价观察干预前、后的患者是否有不良反应以及膝关节活动度的评分和疼痛视觉模拟评分法(VAS)评分,以及核磁共振下膝关节面影像变化。结果:ADSCs膜片移植在兔膝关节股骨髁部软骨损伤的大体照呈透明状,触摸较硬,甲苯胺蓝染色显示有透明软骨组织生成。人腹部脂肪组织分离出来的hADSCs均阳性表达CD105、CD29,阴性表达CD45、CD34,体外具有良好的成骨、成脂、成软骨能力;临床治疗干预后患者无不良反应,膝疼痛减轻,关节活动度较前增大,核磁共振可见关节面较治疗前斑片状影较治疗前显著减少,低信号区显著减少,T2值减低,未见明显关节腔积液,关节面厚度增加。结论:ADSCs在兔关节内环境作用下可被诱导分化成透明软骨组织。ADSCs可以作为软骨缺损修复或者骨性关节炎修复的种子细胞。关节腔注射hADSCs复合玻璃酸钠、富血小板血浆(PRP)治疗骨性关节炎能有效改善患者膝关节活动度,减少疼痛,提高生活质量;通过核磁共振检查发现关节面厚度增加,原软骨损伤部位显示修复影像。采用规范的关节穿刺注射技术安全可靠,临床适用性强。此方法安全有效,可以延缓关节退化,提高患者生活质量。
[Abstract]:Aim: to investigate the effect of allogeneic ADSCs cell membrane as seed cell for repairing cartilage defect and to induce growth and differentiation into hyaline cartilage tissue under the action of rabbit knee joint cavity microenvironment. Safety and efficacy of injection of autologous adipose-derived mesenchymal stem cells (Human adipose derived mesenchymal stem cells,hADSCs) combined with sodium hyaluronate and platelet-rich plasma (platelet rich plasma,PRP) in the treatment of knee osteoarthritis (Osteoarthritis,OA). Methods: experiment 1: (1) adipose tissue of New Zealand white rabbits was isolated and cultured. ADSCs membrane was prepared. (2) New Zealand white rabbit knee joint femoral condylar cartilage injury model was transplanted with ADSCs membrane. (3) ADSC repair rabbit cartilage injury large body radiography and histological evaluation. Experiment 2: (1) Human abdominal adipose tissue was isolated from human abdominal adipose tissue, human ADSCs, was isolated and cultured in vitro to obtain a sufficient amount of ADSCS, and its surface markers were detected by flow cytometry. The differentiation ability of chondrocytes was analyzed. (2) 30 patients with mild to moderate osteoarthritis of knee joint were selected from the outpatient department of orthopedics center of first people's Hospital of Xianyang City. Human ADSCs combined with sodium hyaluronate was injected into articular cavity and platelet-rich plasma (PRP), was injected into the joint. To evaluate whether the patients before and after intervention had adverse reactions, knee motion score and pain visual analogue score (VAS), as well as the changes of knee joint surface image under magnetic resonance imaging (MRI). Results: ADSCs membrane transplantation in rabbit knee joint femoral condylar cartilage injury of the large body is transparent, hard touch, toluene blue staining showed hyaline cartilage tissue formation. The hADSCs positive expression CD105,CD29, negative expression CD45,CD34, in human abdominal adipose tissue had good osteogenic, lipogenic and chondrogenic ability in vitro. No adverse reaction was found in the patients after clinical intervention, the knee pain was relieved, and the motion of the joint was increased. MRI showed that the patchy shadow of the articular surface decreased significantly, the low signal area decreased significantly, and the T _ 2 value decreased significantly compared with the pre-treatment patchy shadow of the joint surface before the treatment. There was no obvious hydrops in the joint cavity, and the thickness of the articular surface increased. Conclusion: ADSCs can be induced to differentiate into hyaline cartilage tissue in rabbit articular environment. ADSCs can be used as seed cells for repair of cartilage defect or repair of osteoarthritis. Intra-articular injection of hADSCs combined with sodium hyaluronate and platelet-rich plasma (PRP) in the treatment of osteoarthritis can effectively improve the knee joint activity, reduce pain, and improve the quality of life. The thickness of articular surface was increased by magnetic resonance imaging (MRI), and the repair images of the original cartilage were displayed. The standard technique of joint puncture injection is safe and reliable, and its clinical applicability is strong. This method is safe and effective and can delay joint degeneration and improve the quality of life of patients.
【学位授予单位】：兰州大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R684.3

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