Utilizing cell-matrix interactions to modulate gene transfer to stem cells inside hyaluronic acid hydrogels.
Utilizing cell-matrix interactions to modulate gene transfer to stem cells inside hyaluronic acid hydrogels.
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DOI：
10.1021/mp200171d
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发表时间：
2011-10-03
影响因子：
4.9
通讯作者：
Segura T
中科院分区：
文献类型：
--
作者：
Gojgini S;Tokatlian T;Segura T
DNA 的有效局部递送将增加基因治疗在组织再生中的适用性，即原位修复患病组织。一种有前途的方法是使用水凝胶支架将质粒DNA以纳米颗粒的形式封装并递送至患病组织，从而使渗透支架的细胞被转染以诱导再生。本研究的重点是负载 DNA 纳米粒子的水凝胶支架的设计。特别是，本研究侧重于了解细胞-基质相互作用如何影响基因转移到基质金属蛋白酶可降解透明质酸水凝胶支架内培养的成体干细胞。使用基质金属蛋白酶可降解肽和迈克尔加成化学将透明质酸交联形成水凝胶材料。这些水凝胶材料内的基因转移被评估为复合物氮磷比（N/P = 5至12）、基质硬度（100至1700 Pa）、RGD浓度（10至400 µM）和RGD呈现（每个HA分子0.2至4.7 RGD）的函数。发现所有变量都会影响 DNA 负载水凝胶内小鼠间充质干细胞培养物的基因转移。正如预期的那样，较高的 N/P 比导致较高的基因转移效率，但毒性也较高，较软的水凝胶比较硬的水凝胶导致更高的转基因表达，中间的 RGD 浓度和 RGD 聚类导致更高的转基因表达。我们相信，通过这种体外模型获得的知识可用于设计更好的支架介导的基因递送，用于局部基因治疗。
The effective delivery of DNA locally would increase the applicability of gene therapy in tissue regeneration, where diseased tissue is to be repaired in situ. One promising approach is to use hydrogel scaffolds to encapsulate and deliver plasmid DNA in the form of nanoparticles to the diseased tissue, so that cells infiltrating the scaffold are transfected to induce regeneration. This study focuses on the design of a DNA nanoparticle loaded hydrogel scaffold. In particular, this study focuses on understanding how cell-matrix interactions affect gene transfer to adult stem cells cultured inside matrix metalloproteinase degradable hyaluronic acid hydrogel scaffolds. Hyaluronic acid was crosslinked to form a hydrogel material using a matrix metalloproteinase degradable peptide and Michael addition chemistry. Gene transfer inside these hydrogel materials was assessed as a function of polyplex nitrogen to phosphate ratio (N/P = 5 to 12), matrix stiffness (100 to 1700 Pa), RGD concentration (10 to 400 µM) and RGD presentation (.2 to 4.7 RGDs per HA molecule). All variables were found to affect gene transfer to mouse mensenchymal stem cells culture inside the DNA loaded hydrogels. As expected higher N/P ratios lead to higher gene transfer efficiency but also higher toxicity, softer hydrogels resulted in higher transgene expression than stiffer hydrogels, an intermediate RGD concentration and RGD clustering resulted in higher transgene expression. We believe that the knowledge gained through this in vitro model can be utilized to design better scaffold-mediated gene delivery for local gene therapy.
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影响因子：
9.7
作者：
Dhaliwal A;Maldonado M;Han Z;Segura T
通讯作者：
Segura T
影响因子：
82.9
作者：
Bonadio, J;Smiley, E;Goldstein, S
通讯作者：
Goldstein, S
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5.1
作者：
Aoki, K;Furuhata, S;Yoshida, T
通讯作者：
Yoshida, T
影响因子：
12.4
作者：
Gautam, A;Densmore, CL;Waldrep, JC
通讯作者：
Waldrep, JC
影响因子：
12.4
作者：
Jang, JH;Rives, CB;Shea, LD
通讯作者：
Shea, LD