背景:人脂肪干细胞(ADSCs)具有很强的抗炎作用,滑膜来源的干细胞(SDSCs)具有很高的软骨形成潜能。因此,这项研究旨在研究人类ADSCs和SDSCs的组合是否会产生协同作用,从而在体外增加骨关节炎(OA)软骨细胞的软骨形成潜能,并在体外减轻早期和晚期OA的软骨退变。
方法:ADSCs,SDSC,从接受全膝关节置换术的OA患者中分离出软骨细胞。ADSCs-SDSCs混合细胞比例为1:0(仅ADSCs),8:2,5:5(5A5S),2:8和0:1(仅SDSC)。用transwell测定法或具有各种混合细胞组的沉淀培养物体外评估OA软骨细胞的软骨形成潜力。然后选择具有最高软骨形成潜能的混合细胞组,并在体内注射到早期和晚期OA阶段的裸大鼠的膝关节中。然后在手术后12周和20周通过步态分析对动物进行评估,VonFrey测试,显微计算机断层扫描,MRI,免疫组织化学和组织学分析。最后,通过体内组织样本的RNA测序和OA软骨细胞自噬途径的Western印迹研究了这些发现的潜在机制.
结果:在MSCs治疗组中,5A5S具有最大的协同作用,在体外增加OA软骨细胞的软骨形成潜力,在体内抑制早期和晚期OA。5A5S组软骨退变明显减轻,滑膜炎症,疼痛感觉,软骨下裸鼠OA的神经侵犯,优于两种单细胞治疗。其潜在机制是通过FoxO1信号通路激活软骨细胞自噬。
结论:人类ADSCs和SDSCs的组合显示出比单一类型干细胞更高的潜力,证明了作为一种新型治疗OA的潜力。
BACKGROUND: Human adipose-derived stem cells (ADSCs) exert a strong anti-inflammatory effect, and synovium-derived stem cells (SDSCs) have high chondrogenic potential. Thus, this study aims to investigate whether a combination of human ADSCs and SDSCs will have a synergistic effect that will increase the chondrogenic potential of osteoarthritis (OA) chondrocytes in vitro and attenuate the cartilage degeneration of early and advanced OA in vitro.
METHODS: ADSCs, SDSCs, and chondrocytes were isolated from OA patients who underwent total knee arthroplasty. The ADSCs-SDSCs mixed cell ratios were 1:0 (ADSCs only), 8:2, 5:5 (5A5S), 2:8, and 0:1 (SDSCs only). The chondrogenic potential of the OA chondrocytes was evaluated in vitro with a transwell assay or pellet culture with various mixed cell groups. The mixed cell group with the highest chondrogenic potential was then selected and injected into the knee joints of nude rats of early and advanced OA stages in vivo. The animals were then evaluated 12 and 20 weeks after surgery through gait analysis, von frey test, microcomputed tomography, MRI, and immunohistochemical and histological analyses. Finally, the mechanisms underlying these findings were investigated through the RNA sequencing of tissue samples in vivo and Western blot of the OA chondrocyte autophagy pathway.
RESULTS: Among the MSCs treatment groups, 5A5S had the greatest synergistic effect that increased the chondrogenic potential of OA chondrocytes in vitro and inhibited early and advanced OA in vivo. The 5A5S group significantly reduced cartilage degeneration, synovial inflammation, pain sensation, and nerve invasion in subchondral nude rat OA, outperforming both single-cell treatments. The underlying mechanism was the activation of chondrocyte autophagy via the FoxO1 signaling pathway.
CONCLUSIONS: A combination of human ADSCs and SDSCs demonstrated higher potential than a single type of stem cell, demonstrating potential as a novel treatment for OA.