WNT3A-loaded exosomes enable cartilage repair.
WNT3A-loaded exosomes enable cartilage repair.
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DOI：
10.1002/jev2.12088
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发表时间：
2021-05
影响因子：
16
通讯作者：
Dell'Accio F
中科院分区：
文献类型：
--
作者：
Thomas BL;Eldridge SE;Nosrati B;Alvarez M;Thorup AS;Nalesso G;Caxaria S;Barawi A;Nicholson JG;Perretti M;Gaston-Massuet C;Pitzalis C;Maloney A;Moore A;Jupp R;Dell'Accio F
软骨缺损修复不良。最近的遗传学研究表明，WNT3a可能有助于软骨再生，然而致密的无血管软骨细胞外基质限制了它的渗透和向软骨细胞发出信号。细胞外囊泡活跃地穿透完整软骨。本研究探讨了使用外泌体作为递送载体在体内将WNT3a递送到大软骨缺损中的效果。外泌体通过超离心从过表达WNT3a的L细胞或对照未转导的L细胞的条件培养基中纯化，并通过电子显微镜、纳米颗粒跟踪分析和标记谱进行表征。负载在外泌体上的WNT3a通过western blotting进行定量，并使用SUPER8TOPFlash报告基因检测和其他已建立的读数（包括增殖和蛋白多糖含量）进行体外功能表征。使用TCF/ left:H2B‐GFP报告小鼠评估体内通路激活。将装载Wnt3a的外泌体注射到小鼠的膝盖中，通过手术产生大的骨软骨缺损。8周后对修复程度进行组织学评分。WNT3a被成功装载到外泌体上，并在体外激活了WNT信号传导。在体内，重组WNT3a不能激活软骨中的WNT信号，而装载WNT3a的外泌体单次激活典型WNT信号至少一周，八周后，改善了骨软骨缺陷的修复。WNT3a组装在外泌体上，有效地递送到软骨中，有助于骨软骨缺损的愈合。
Cartilage defects repair poorly. Recent genetic studies suggest that WNT3a may contribute to cartilage regeneration, however the dense, avascular cartilage extracellular matrix limits its penetration and signalling to chondrocytes. Extracellular vesicles actively penetrate intact cartilage. This study investigates the effect of delivering WNT3a into large cartilage defects in vivo using exosomes as a delivery vehicle. Exosomes were purified by ultracentrifugation from conditioned medium of either L‐cells overexpressing WNT3a or control un‐transduced L‐cells, and characterized by electron microscopy, nanoparticle tracking analysis and marker profiling. WNT3a loaded on exosomes was quantified by western blotting and functionally characterized in vitro using the SUPER8TOPFlash reporter assay and other established readouts including proliferation and proteoglycan content. In vivo pathway activation was assessed using TCF/Lef:H2B‐GFP reporter mice. Wnt3a loaded exosomes were injected into the knees of mice, in which large osteochondral defects were surgically generated. The degree of repair was histologically scored after 8 weeks. WNT3a was successfully loaded on exosomes and resulted in activation of WNT signalling in vitro. In vivo, recombinant WNT3a failed to activate WNT signalling in cartilage, whereas a single administration of WNT3a loaded exosomes activated canonical WNT signalling for at least one week, and eight weeks later, improved the repair of osteochondral defects. WNT3a assembled on exosomes, is efficiently delivered into cartilage and contributes to the healing of osteochondral defects.
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DOI：
10.1053/jars.2002.32839
发表时间：
2002-09-01
影响因子：
4.7
作者：
Hjelle, K;Solheim, E;Brittberg, M
通讯作者：
Brittberg, M
DOI：
10.1016/s0749-8063(97)90124-9
发表时间：
1997-08-01
影响因子：
4.7
作者：
Curl, WW;Krome, J;Poehling, GG
通讯作者：
Poehling, GG
影响因子：
7
作者：
Eltawil NM;De Bari C;Achan P;Pitzalis C;Dell'accio F
通讯作者：
Dell'accio F
影响因子：
56.9
作者：
Johnson, Kristen;Zhu, Shoutian;Schultz, Peter G.
通讯作者：
Schultz, Peter G.
影响因子：
16.6
作者：
Monteagudo S;Cornelis FMF;Aznar-Lopez C;Yibmantasiri P;Guns LA;Carmeliet P;Cailotto F;Lories RJ
通讯作者：
Lories RJ