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首页> 外文期刊>Acta biomaterialia >Design and characterization of microporous hyaluronic acid hydrogels for in vitro gene transfer to mMSCs.
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Design and characterization of microporous hyaluronic acid hydrogels for in vitro gene transfer to mMSCs.

机译：用于体外基因转移至mMSCs的微孔透明质酸水凝胶的设计和表征。

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摘要

The effective and sustained delivery of DNA locally could increase the applicability of gene therapy in tissue regeneration and therapeutic angiogenesis. One promising approach is to use porous hydrogel scaffolds to encapsulate and deliver nucleotides in the form of nanoparticles to the affected sites. We have designed and characterized microporous (μ-pore) hyaluronic acid hydrogels which allow for effective cell seeding in vitro post-scaffold fabrication and allow for cell spreading and proliferation without requiring high levels of degradation. These factors, coupled with high loading efficiency of DNA polyplexes using a previously developed caged nanoparticle encapsulation (CnE) technique, then allowed for long-term sustained transfection and transgene expression of incorporated mMSCs. In this study, we examined the effect of pore size on gene transfer efficiency and the kinetics of transgene expression. For all investigated pore sizes (30, 60, and 100 μm), encapsulated DNA polyplexes were released steadily, starting by day 4 for up to 10 days. Likewise, transgene expression was sustained over this period, although significant differences between different pore sizes were not observed. Cell viability was also shown to remain high over time, even in the presence of high concentrations of DNA polyplexes. The knowledge acquired through this in vitro model can be utilized to design and better predict scaffold-mediated gene delivery for local gene therapy in an in vivo model where host cells infiltrate the scaffold over time.
机译：DNA的有效和持续传递可以增加基因治疗在组织再生和治疗性血管生成中的适用性。一种有前途的方法是使用多孔水凝胶支架将纳米颗粒形式的核苷酸包封并递送至患处。我们已经设计并表征了微孔（μ孔）透明质酸水凝胶，可在体外支架制造后进行有效的细胞接种，并允许细胞扩散和增殖而无需高水平的降解。这些因素，再加上使用先前开发的笼式纳米粒子封装（CnE）技术的DNA多链体的高加载效率，可实现所整合mMSC的长期持续转染和转基因表达。在这项研究中，我们检查了孔径对基因转移效率和转基因表达动力学的影响。对于所有调查的孔径（30、60和100μm），封装的DNA多链体稳定释放，从第4天开始长达10天。同样，尽管未观察到不同孔径之间的显着差异，但转基因表达在此期间得以维持。即使存在高浓度的DNA多链体，细胞活力也随着时间的推移而保持高水平。通过该体外模型获得的知识可用于设计和更好地预测在宿主细胞随时间渗入支架的体内模型中用于局部基因治疗的支架介导的基因递送。

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