In vivo delivery of functional Flightless I siRNA using layer-by-layer polymer surface modification

J Biomater Appl. 2015 Sep;30(3):257-68. doi: 10.1177/0885328215579422. Epub 2015 Apr 2.

Abstract

Gene silencing using small interfering RNA has been proposed as a therapy for cancer, viral infections and other diseases. This study aimed to investigate whether layer-by-layer polymer surface modification could deliver small interfering RNA to decrease fibrotic processes associated with medical device implantation. Anti-green fluorescent protein labelled small interfering RNA was applied to tissue culture plates and polyurethane using a layer-by-layer technique with small interfering RNA and poly-L-lysine. In vitro studies showed that the level of down-regulation of green fluorescent protein was directly related to the number of coatings applied. This layer-by-layer coating technique was then used to generate Rhodamine-Flii small interfering RNA-coated implants for in vivo studies of small interfering RNA delivery via subcutaneous implantation in mice. After two days, Rh-positive cells were observed on the implants' surface indicating cellular uptake of the Rhodamine-Flii small interfering RNA. Decreased Flii gene expression was observed in tissue surrounding the Rhodamine-Flii small interfering RNA coated implants for up to seven days post implantation, returning to baseline by day 21. Genes downstream from Flii, including TGF-β1 and TGF-β3, showed significantly altered expression confirming a functional effect of the Rhodamine-Flii small interfering RNA on gene expression. This research demonstrates proof-of-principle that small interfering RNA can be delivered via layer-by-layer coatings on biomaterials and thereby can alter the fibrotic process.

Keywords: Animal model; drug delivery; polyurethane; siRNA.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Biocompatible Materials*
  • Carrier Proteins
  • Cell Line
  • Cytoskeletal Proteins / genetics*
  • Mice
  • Mice, Inbred BALB C
  • Microfilament Proteins
  • NIH 3T3 Cells
  • Polymers / chemistry*
  • RNA, Small Interfering / administration & dosage*
  • Surface Properties
  • Trans-Activators

Substances

  • Biocompatible Materials
  • Carrier Proteins
  • Cytoskeletal Proteins
  • FlII protein, mouse
  • Microfilament Proteins
  • Polymers
  • RNA, Small Interfering
  • Trans-Activators