Supramolecular polypseudorotaxane gels for controlled delivery of rAAV vectors in human mesenchymal stem cells for regenerative medicine.
Supramolecular polypseudorotaxane gels for controlled delivery of rAAV vectors in human mesenchymal stem cells for regenerative medicine.
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DOI：
10.1016/j.ijpharm.2017.05.050
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发表时间：
2017-10
影响因子：
5.8
通讯作者：
A. Rey-Rico;Heiko Babicz;H. Madry;A. Concheiro;C. Alvarez‐Lorenzo;M. Cucchiarini
中科院分区：
文献类型：
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作者：
A. Rey-Rico;Heiko Babicz;H. Madry;A. Concheiro;C. Alvarez‐Lorenzo;M. Cucchiarini
这项工作的目的是调查，第一次，使用超分子polypropylodorotaxane凝胶作为支架，可以持久地提供rAAV载体的应用在软骨再生的可能性。在PBS中制备含有透明质酸（HA）或硫酸软骨素（CS）的Pluronic®F68（PF 68）或Tetronic®908（T908）的分散体。然后将α-环糊精（αCD）加入到一定的分散体中形成聚β-多西紫杉烷凝胶。在体外研究了rAAV载体从释放介质和直接接触凝胶中的释放以及rAAV载体对人骨髓间充质干细胞（hMSCs）的转导。与游离载体相比，凝胶提供了更高水平的转基因表达。CS（或HA）/PF 68/αCD凝胶可快速释放rAAV载体，而CS（或HA）/T908/αCD凝胶可持续释放rAAV载体，这可能与其与病毒载体的相互作用不同有关。将αCD掺入CS（或HA）/PF 68凝胶中导致rAAV浓度升高，并随时间推移维持转基因表达水平。HA提高了凝胶的生物活性和细胞相容性，特别是基于T908的凝胶。总的来说，结合rAAV基因转移与polypeptide dorotaxane凝胶可能为人类组织工程和再生医学策略提供新的，有前途的工具。
The aim of this work was to investigate, for the first time, the possibility of using supramolecular polypseudorotaxane gels as scaffolds that can durably deliver rAAV vectors for applications in cartilage regeneration. Dispersions of Pluronic®F68 (PF68) or Tetronic®908 (T908) containing either hyaluronic acid (HA) or chondroitin sulfate (CS) were prepared in PBS. Then, alpha-cyclodextrin (αCD) was added to some dispersions to form polypseudorotaxane gels. Polysaccharides and αCD reinforced the viscoelasticity of the gels, which could withstand autoclaving without changes.In vitrorelease of rAAV vectors and subsequent transduction of human mesenchymal stem cells (hMSCs) by rAAV vectors from the release medium and from gels in direct contact with the cells were investigated. Compared with free vectors, the gels provided higher levels of transgene expression. CS (or HA)/PF68/αCD gels rapidly released rAAV vectors while CS (or HA)/T908/αCD gels provided sustained release probably due to different interactions with the viral vectors. Incorporation of αCD into CS (or HA)/PF68 gels resulted on higher rAAV concentrations and sustained levels of transgene expression over time. HA increased the bioactivity and cytocompatibility of the gels, especially those based on T908. Overall, combining rAAV gene transfer with polypseudorotaxane gels may provide new, promising tools for human tissue engineering and regenerative medicine strategies.