Jing Xu-hong, Yang Liu, Duan Xiao-jun, Xie Bing, Chen Wei, Li Zhong, Tan Hong-bo
Center of Joint Surgery, Southwest Hospital, The Third Military Medical University, Chongqing, China.
Joint Bone Spine. 2008 Jul;75(4):432-8. doi: 10.1016/j.jbspin.2007.09.013. Epub 2008 May 2.
To track superparamagnetic iron oxide nanoparticle (SPIO)-labeled, bone-derived mesenchymal stem cells (MSCs) by in vivo magnetic resonance imaging (MRI) with a 1.5T-system following injection of engineered autologous MSCs into the knee joint cavity in rabbit articular cartilage defect models.
Rabbit MSCs were labeled with SPIO and a transfection agent. Cell viability, proliferation and differentiation capacity were assessed in vitro using appropriate functional assays. Cells underwent GRE T2*-weighted MRI in vitro. The autologous MSCs seeded in chitosan and glycerophosphate (C-GP) gel were injected into the knee joint cavity of rabbit models for cartilage defects. All rabbits underwent GRE T2*-weighted MRI 1, 4, 8 and 12 weeks post-injection. MR imaging findings were compared histologically.
Nanoparticles were stained with Prussian blue and observed by transmission electron microscopy inside the cells. Cell viability, proliferation, and differentiation were comparable between labeled and non-labeled cells. After intra-articular injection of labeled autologous MSCs, marked hypointense signal void areas representing the injected MSCs can be observed for at least 12 weeks on GRE T2*-weighted images. At 12 weeks post-injection, labeled MSCs migrated into the synovial fluid at the suprapatellar bursa, the popliteal space site and subchondral bone of the femur but no MSCs were detected in the defect. Histochemical staining confirmed the presence of Prussian blue-positive cells and BrdU-positive cells.
MRI would be an efficient noninvasive technique to visually track SPIO-labeled seed cells in vivo; the engineered autologous MSCs do not actively participate in the repair of articular cartilage defects following intra-articular injection.
在兔关节软骨缺损模型的膝关节腔注射工程化自体间充质干细胞(MSCs)后,利用1.5T系统通过体内磁共振成像(MRI)追踪超顺磁性氧化铁纳米颗粒(SPIO)标记的骨源性间充质干细胞。
用SPIO和转染剂标记兔MSCs。使用适当的功能测定法在体外评估细胞活力、增殖和分化能力。细胞在体外进行GRE T2加权MRI检查。将接种于壳聚糖和甘油磷酸(C-GP)凝胶中的自体MSCs注入兔软骨缺损模型的膝关节腔。所有兔子在注射后1、4、8和12周接受GRE T2加权MRI检查。将MR成像结果与组织学结果进行比较。
纳米颗粒用普鲁士蓝染色,并通过透射电子显微镜在细胞内观察到。标记细胞和未标记细胞之间的细胞活力、增殖和分化相当。关节内注射标记的自体MSCs后,在GRE T2*加权图像上至少12周可观察到代表注射MSCs的明显低信号空洞区域。注射后12周,标记的MSCs迁移到髌上囊、腘窝间隙部位的滑液和股骨的软骨下骨,但在缺损处未检测到MSCs。组织化学染色证实存在普鲁士蓝阳性细胞和BrdU阳性细胞。
MRI将是一种在体内直观追踪SPIO标记的种子细胞的有效非侵入性技术;关节内注射后,工程化自体MSCs不积极参与关节软骨缺损的修复。