Device-based local delivery of siRNA against mammalian target of rapamycin (mTOR) in a murine subcutaneous implant model to inhibit fibrous encapsulation.
Fibrous encapsulation of surgically implanted devices is associated with elevated proliferation and activation of fibroblasts in tissues surrounding these implants, frequently causing foreign body complications. Here we test the hypothesis that inhibition of the expression of mammalian target of rapamycin (mTOR) in fibroblasts can mitigate the soft tissue implant foreign body response by suppressing fibrotic responses around implants. In this study, mTOR was knocked down using small interfering RNA (siRNA) conjugated with branched polyethylenimine (bPEI) in fibroblastic lineage cells in serum-based cell culture as shown by both gene and protein analysis. This mTOR knock-down led to an inhibition in fibroblast proliferation by 70% and simultaneous down-regulation in the expression of type I collagen in fibroblasts in vitro. These siRNA/bPEI complexes were released from poly(ethylene glycol) (PEG)-based hydrogel coatings surrounding model polymer implants in a subcutaneous rodent model in vivo. No significant reduction in fibrous capsule thickness and mTOR expression in the foreign body capsules were observed. The siRNA inefficacy in this in vivo implant model was attributed to siRNA dosing limitations in the gel delivery system, and lack of targeting ability of the siRNA complex specifically to fibroblasts. While in vitro data supported mTOR knock-down in fibroblast cultures, in vivo siRNA delivery must be further improved to produce clinically relevant effects on fibrotic encapsulation around implants.
Takahashi
Hironobu
H
Department of Pharmaceutics and Pharmaceutical Chemistry, University of Utah, Salt Lake City, UT 84112-5820, USA. htakahashi@abmes.twmu.ac.jp
Wang
Yuwei
Y
Grainger
David W
DW
D000818:Animals; D049109:Cell Proliferation; D024042:Collagen Type I / Q000235:genetics / Q000378:metabolism; D015536:Down-Regulation; D004343:Drug Implants / Q000009:adverse effects*; D005260:Female; D005347:Fibroblasts / Q000378:metabolism* / Q000473:pathology; D005355:Fibrosis; D005549:Foreign-Body Reaction / Q000209:etiology / Q000235:genetics / Q000473:pathology / Q000517:prevention & control*; D020100:Hydrogels; D007700:Kinetics; D051379:Mice; D008810:Mice, Inbred C57BL; D041681:NIH 3T3 Cells; D011092:Polyethylene Glycols / Q000737:chemistry; D011094:Polyethyleneimine / Q000737:chemistry; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000008:administration & dosage* / Q000737:chemistry / Q000378:metabolism; D012995:Solubility; D058570:TOR Serine-Threonine Kinases / Q000235:genetics* / Q000378:metabolism
D024042:Collagen Type I; D004343:Drug Implants; D020100:Hydrogels; D034741:RNA, Small Interfering; D011092:Polyethylene Glycols; D011094:Polyethyleneimine; C546843:mTOR protein, mouse; D058570:TOR Serine-Threonine Kinases