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首页> 外文期刊>Biomaterials >The use of gene activated matrix to mediate effective SMAD2 gene silencing against hypertrophic scar
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The use of gene activated matrix to mediate effective SMAD2 gene silencing against hypertrophic scar

机译:基因激活基质在介导针对肥厚性瘢痕的有效SMAD2基因沉默中的应用

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Hypertrophic scar (HS) originates from the over-expression of transforming growth factor β (TGF-β) and downstream SMAD2. With attempts to rectify HS by RNA interference (RNAi) against SMAD2, we report the design of plasmid DNA encoding SMAD2 siRNA (pSUPER-SMAD2), and identify the optimal siRNA sequence toward maximal RNAi efficiency. To realize effective and sustained RNAi, we developed gene activated matrix (GAM) based on porous atelocollagen scaffold and embedded trimethyl chitosan-cysteine (TMCC)/pSUPER-SMAD2 polyplexes for promoting cell growth and gene transfection. The GAM exhibited porosity higher than 80%, pore size of 200-250μm, desired mechanical strength, and sustained pSUPER-SMAD2 release profiles. Normal skin fibroblasts (NSFs) and hypertrophic scar fibroblasts (HSFs) were allowed to infiltrate and proliferate in GAM; at the meantime they were transfected with TMCC/pSUPER-SMAD2 polyplexes to display remarkably reduced SMAD2 levels that lasted for up to 10 days, consequently inhibiting the over-production of type I and type III collagen. We further unraveled the notably higher transfection levels of GAM in three-dimensional (3D) than in 2D environment, which was attributed to the improved cell-matrix interactions that promote cell proliferation and polyplex internalization. This highly safe and effective GAM may serve as a promising candidate towards HS treatment.
机译:肥厚性瘢痕(HS)源自转化生长因子β(TGF-β)和下游SMAD2的过度表达。通过尝试通过针对SMAD2的RNA干扰(RNAi)纠正HS,我们报告了编码SMAD2 siRNA(pSUPER-SMAD2)的质粒DNA的设计,并确定了朝向最大RNAi效率的最佳siRNA序列。为了实现有效和持续的RNAi,我们开发了基于多孔胶原蛋白支架和嵌入式三甲基壳聚糖-半胱氨酸(TMCC)/ pSUPER-SMAD2多聚体的基因激活基质(GAM),以促进细胞生长和基因转染。 GAM表现出高于80%的孔隙率,200-250μm的孔径,所需的机械强度以及持续的pSUPER-SMAD2释放曲线。正常皮肤成纤维细胞(NSFs)和肥厚性瘢痕成纤维细胞(HSFs)被允许在GAM中浸润和增殖;同时,将它们用TMCC / pSUPER-SMAD2多聚体转染以显示SMAD2水平显着降低,可持续长达10天,从而抑制了I型和III型胶原蛋白的过度生产。我们进一步揭示了GAM在3D(3D)中的转染水平明显高于2D环境,这归因于改善的细胞-基质相互作用,促进了细胞增殖和多聚体内在化。这种高度安全和有效的GAM可能成为HS治疗的有希望的候选者。

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