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首页> 美国卫生研究院文献>other >Controlling fibrous capsule formation through long-term down-regulation of collagen type I (COL1A1) expression by nanofiber-mediated siRNA gene silencing
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Controlling fibrous capsule formation through long-term down-regulation of collagen type I (COL1A1) expression by nanofiber-mediated siRNA gene silencing

机译：通过纳米纤维介导的siRNA沉默的长期下调胶原型I（COL1A1）表达的长期下调纤维状胶囊形成

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摘要

The foreign body reaction often interferes with the long-term functionality and performance of implanted biomedical devices through fibrous capsule formation. While many implant modification techniques have been adopted in attempts to control fibrous encapsulation, the outcomes remained sub-optimal. Nanofiber scaffold-mediated RNA interference may serve as an alternative approach through the localized and sustained delivery of siRNA at implant sites. In this study, we investigated the efficacy of siRNA-PCLEEP (poly(caprolactone-co-ethylethylene phosphate) nanofibers in controlling fibrous capsule formation through the down-regulation of Collagen type I (COL1A1) in vitro and in vivo. By encapsulating complexes of COL1A1 siRNA with a transfection reagent (Transit TKO) or cell penetrating peptides (CPPs), CADY or MPG, within the nanofibers (550–650 nm in diameter), a sustained release of siRNA was obtained for at least 28 days (loading efficiency ~ 60–67%). Scaffold-mediated transfection significantly enhanced cellular uptake of oligonucleotides and prolonged in vitro gene silencing duration by at least 2–3 times as compared to conventional bolus delivery of siRNA (14 days vs 5–7 days by bolus delivery). In vivo subcutaneous implantation of siRNA scaffolds revealed a significant decrease in fibrous capsule thickness at weeks 2 and 4 as compared to plain nanofibers (p < 0.05). Taken together, the results demonstrated the efficacy of scaffold-mediated siRNA gene-silencing in providing effective long-term control of fibrous capsule formation.
机译：异物反应通常会通过纤维囊的形成而干扰植入式生物医学设备的长期功能和性能。虽然已采用许多植入物修饰技术来尝试控制纤​​维包囊，但结果仍然欠佳。纳米纤维支架介导的RNA干扰可通过在植入位点局部和持续递送siRNA来作为替代方法。在这项研究中，我们研究了siRNA-PCLEEP（聚（己内酯-乙基乙基磷酸乙烯酯）纳米纤维）在体外和体内通过下调I型胶原（COL1A1）来控制纤维囊形成的功效。在纳米纤维（直径为550-650 nm）内，将COL1A1 siRNA与转染试剂（Transit TKO）或细胞穿透肽（CPP），CADY或MPG结合使用，可连续释放siRNA至少28天（负载效率〜 （60–67％）。与传统的单次推注siRNA相比，支架介导的转染显着提高了寡核苷酸的细胞摄取，并延长了体外基因沉默时间至少2-3倍（14 d vs 5–7 d） 。与普通的纳米纤维相比，在体内皮下植入siRNA支架显示在第2和第4周纤维囊厚度显着降低（p <0.05）。 affold介导的siRNA基因沉默可有效地长期控制纤维囊的形成。

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