Spatial control of gene expression within a scaffold by localized inducer release.
Spatial control of gene expression within a scaffold by localized inducer release.
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DOI：
10.1016/j.biomaterials.2010.12.037
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发表时间：
2011-04
期刊：
影响因子：
14
通讯作者：
Wagner, William R.
中科院分区：
文献类型：
--
作者：
Baraniak, Priya R.;Nelson, Devin M.;Leeson, Cory E.;Katakam, Anand K.;Friz, Jennifer L.;Cress, Dean E.;Hong, Yi;Guan, Jianjun;Wagner, William R.
基因表达可以通过使用诱导物/启动子系统在遗传修饰的细胞中控制，其中诱导物分子的存在调节基因表达的时间和水平。通过应用控制释放的原理，应该可以通过诱导物从下面的材料基质中释放的存在或不存在来控制生物材料表面上的基因表达，从而避免依赖于从材料表面摄取相对不稳定的DNA的替代技术。为了评估这一概念，一个修改的蜕皮激素响应基因表达系统转染到B16小鼠细胞和诱导配体，这是从弹性体聚（酯氨基甲酸酯）脲（PEUU）释放的能力，启动基因表达进行了研究。首先将合成的诱导配体加载到PEUU中，以证明在体外一年内在各种加载密度下生物活性分子的延长释放。图案化膜的PEUU可变加载诱导剂导致在空间上控制细胞表达的基因产物（绿色荧光蛋白，GFP）。在由PEUU通过盐浸提制成的多孔支架中，其中中心区域专门装载有诱导剂，细胞主要在装载的中心区域表达GFP，而在省略配体的外部区域表达最小。这种支架系统最终可以提供一种在体内以空间限定的方式精确控制祖细胞定型的方法，用于软组织修复和再生。
Gene expression can be controlled in genetically modified cells by employing an inducer/promoter system where presence of the inducer molecule regulates the timing and level of gene expression. By applying the principles of controlled release, it should be possible to control gene expression on a biomaterial surface by the presence or absence of inducer release from the underlying material matrix, thus avoiding alternative techniques that rely upon uptake of relatively labile DNA from material surfaces. To evaluate this concept, a modified ecdysone-responsive gene expression system was transfected into B16 murine cells and the ability of an inducer ligand, which was released from elastomeric poly(ester urethane) urea (PEUU), to initiate gene expression was studied. The synthetic inducer ligand was first loaded into PEUU to demonstrate extended release of the bioactive molecule at various loading densities over a one year period in vitro. Patterning films of PEUU variably-loaded with inducer resulted in spatially controlled cell expression of the gene product (green fluorescent protein, GFP). In porous scaffolds made from PEUU by salt leaching, where the central region was exclusively loaded with inducer, cells expressed GFP predominately in the loaded central regions whereas expression was minimal in outer regions where ligand was omitted. This scaffold system may ultimately provide a means to precisely control progenitor cell commitment in a spatially-defined manner in vivo for soft tissue repair and regeneration.
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影响因子：
14
作者：
Hashizume, Ryotaro;Fujimoto, Kazuro L.;Hong, Yi;Amoroso, Nicholas J.;Tobita, Kimimasa;Miki, Toshio;Keller, Bradley B.;Sacks, Michael S.;Wagner, William R.
通讯作者：
Wagner, William R.
DOI：
10.1002/jbm.10204
发表时间：
2002-09-05
期刊：
JOURNAL OF BIOMEDICAL MATERIALS RESEARCH
影响因子：
--
作者：
Guan, JJ;Sacks, MS;Wagner, WR
通讯作者：
Wagner, WR
影响因子：
4.6
作者：
Campbell, Phil G.;Weiss, Lee E.
通讯作者：
Weiss, Lee E.
影响因子：
5.3
作者：
Kennedy, Timothy E.;Wang, Hao;Tessier-Lavigne, Marc
通讯作者：
Tessier-Lavigne, Marc
影响因子：
4.3
作者：
Baker, R. E.;Maini, P. K.
通讯作者：
Maini, P. K.