Regulated non-viral gene delivery from coaxial electrospun fiber mesh scaffolds.
Regulated non-viral gene delivery from coaxial electrospun fiber mesh scaffolds.
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DOI：
10.1016/j.jconrel.2009.12.009
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发表时间：
2010-04-02
影响因子：
10.8
通讯作者：
Mikos, Antonios G.
中科院分区：
文献类型：
--
作者：
Saraf, Anita;Baggett, L. Scott;Raphael, Robert M.;Kasper, F. Kurtis;Mikos, Antonios G.
为了增加三维支架在组织工程应用中的多功能性，最近的实验设计将生物分子如质粒和蛋白质掺入支架结构中。这样的支架充当感兴趣的生物分子的储库，同时在不同的持续时间内调节它们的释放。在这里，我们描述了使用同轴静电纺丝作为一种手段，用于制造纤维网支架和封装和随后释放的非病毒基因递送载体在长达60天的时间。基于部分析因设计，在两个水平上研究了四个处理参数的影响，制造了在芯内含有质粒DNA（pDNA）和在同轴纤维鞘内含有非病毒基因递送载体聚（乙烯亚胺）-透明质酸（PEI-HA）的各种纤维网支架。研究了聚（ε-己内酯）鞘聚合物浓度、聚（乙二醇）核聚合物分子量和浓度以及pDNA浓度对平均纤维直径、PEI-HA释放动力学和转染效率的影响。经确定，增加每个研究参数的值会导致纤维平均直径的增加。PEI-HA从纤维的释放动力学受pDNA的负载浓度影响（相应地调节PEI-HA浓度以维持复合物内恒定的氮磷（N：P）比）。模型成纤维样细胞的二维细胞培养实验表明，从纤维网支架释放的PEI-HA与pDNA的复合物可以成功地转染细胞并诱导增强型绿色荧光蛋白（EGFP）的表达。峰值EGFP表达随所研究的处理参数而变化，并且观察到的平均转染是聚（乙二醇）（核心）分子量和浓度的函数。此外，直接接种到含有PEI-HA和pDNA的同轴纤维网支架上的成纤维细胞样细胞在60天内显示出EGFP表达，这显著大于用仅含有pDNA的支架观察到的EGFP表达。因此，在从电纺同轴纤维网支架释放pDNA和非病毒基因递送载体时，可以在延长的时间段内实现可变的转染活性，其中释放和随后的转染由可调同轴纤维网制造参数控制。
In an effort to add to the versatility of three-dimensional scaffolds for tissue engineering applications, recent experimental designs are incorporating biological molecules such as plasmids and proteins within the scaffold structure. Such scaffolds act as reservoirs for the biological molecules of interest while regulating their release over various durations of time. Here, we describe the use of coaxial electrospinning as a means for the fabrication of fiber mesh scaffolds and the encapsulation and subsequent release of a non-viral gene delivery vector over a period of up to 60 days. Various fiber mesh scaffolds containing plasmid DNA (pDNA) within the core and the non-viral gene delivery vector poly(ethylenimine)-hyaluronic acid (PEI-HA) within the sheath of coaxial fibers were fabricated based on a fractional factorial design that investigated the effects of four processing parameters at two levels. Poly(ε-caprolactone) sheath polymer concentration, poly(ethylene glycol) core polymer molecular weight and concentration, and the concentration of pDNA were investigated for their effects on average fiber diameter, release kinetics of PEI-HA, and transfection efficiency. It was determined that increasing the values of each of the investigated parameters caused an increase in the average diameter of the fibers. The release kinetics of PEI-HA from the fibers were affected by the loading concentration of pDNA (with PEI-HA concentration adjusted accordingly to maintain a constant nitrogen to phosphorous (N:P) ratio within the complexes). Two-dimensional cell culture experiments with model fibroblast-like cells demonstrated that complexes of pDNA with PEI-HA released from fiber mesh scaffolds could successfully transfect cells and induce expression of enhanced green fluorescent protein (EGFP). Peak EGFP expression varied with the investigated processing parameters, and the average transfection observed was a function of poly(ethylene glycol) (core) molecular weight and concentration. Furthermore, fibroblast-like cells seeded directly onto coaxial fiber mesh scaffolds containing PEI-HA and pDNA showed EGFP expression over 60 days, which was significantly greater than the EGFP expression observed with scaffolds containing pDNA alone. Hence, variable transfection activity can be achieved over extended periods of time upon release of pDNA and non-viral gene delivery vectors from electrospun coaxial fiber mesh scaffolds, with release and subsequent transfection controlled by tunable coaxial fiber mesh fabrication parameters.
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DOI：
10.1089/ten.tec.2008.0422
发表时间：
2009-09
期刊：
Tissue engineering. Part C, Methods
影响因子：
--
作者：
Saraf A;Lozier G;Haesslein A;Kasper FK;Raphael RM;Baggett LS;Mikos AG
通讯作者：
Mikos AG
影响因子：
4.6
作者：
Shin, YM;Hohman, MM;Rutledge, GC
通讯作者：
Rutledge, GC
影响因子：
10.8
作者：
Luu, YK;Kim, K;Hadjiargyrou, M
通讯作者：
Hadjiargyrou, M
影响因子：
4.2
作者：
Huang, YC;Riddle, K;Mooney, DJ
通讯作者：
Mooney, DJ
影响因子：
4.9
作者：
Peng, Lin;Cheng, Xiangrong;Li, Siqun
通讯作者：
Li, Siqun