MMP-9 Downregulation with Lipid Nanoparticles for Inhibiting Corneal Neovascularization by Gene Silencing

Abstract

Gene silencing targeting proangiogenic factors have been shown to be a useful strategy in the treatment of corneal neovascularization (CNV). Among interference RNA (RNAi) molecules, short-hairpin RNA (shRNA) is a plasmid-coded RNA able to down-regulate the expression of the desired gene. It is continuously produced in the host cell, inducing a durable gene silencing effect. The aim of this work was to develop a solid lipid nanoparticle (SLN)-based shRNA delivery system to downregulate metalloproteinase 9 (MMP-9), a proangiogenic factor, in corneal cells for the treatment of CNV associated with inflammation. The nanovectors were prepared using a solvent emulsification-evaporation technique, and after physicochemical evaluation, they were evaluated in different culture cell models. Transfection efficacy, cell internalization, cell viability, the effect on MMP-9 expression, and cell migration were evaluated in human corneal epithelial cells (HCE-2). The inhibition of tube formation using human umbilical vein endothelial cells (HUVEC) was also assayed. The non-viral vectors based on SLN were able to downregulate the MMP-9 expression in HCE-2 cells via gene silencing, and, consequently, to inhibit cell migration and tube formation. These results demonstrate the potential of lipid nanoparticles as gene delivery systems for the treatment of CNV-associated inflammation by RNAi technology.

Development of Brain Targeting Peptide Based MMP-9 Inhibiting Nanoparticles for the Treatment of Brain Diseases with Elevated MMP-9 Activity

“…The MMP-9 inhibiting NPs in this work, up to 250 µg/mL, did not present significant toxicity towards HeLa cells, and other nanoparticle formulations also showed similar findings; 17,52,53 while PVP-coated platinum NPs did show cell toxicity at 100 µg/mL concentration. 53 Furthermore, magnetic NPs-bound TMP-1 NPs showed cell toxicity at concentrations above 10 µg/mL.…”

“…In addition, solid lipid NPs-encapsulating MMP-9 downregulating shRNA-reduced the secretion of MMP-9 in human corneal epithelial cells by 30%. 52 The ability of our NPs to inhibit MMP-9 was investigated through the use of a specially-designed and optimised 19 F NMR based assay that measures the remaining 19 F-labelled peptide upon treatment with MMP-9 compared with control. MMP-9 inhibiting NPs of this study had concentration of 480 µM and they reduced the activity of MMP-9 completely or by half.…”

“…16,17 Amongst the nanoparticle formulations discussed above, the PVP stabilised gold NPs, 53 CuS@mSiO2-PEG NPs, 48 and MMP-9 inhibiting NPs of our work did not require loading of an active ingredient, such as TIMP-1. [16][17][18]52 Therefore, the MMP-9 inhibiting NPs of our study would have fewer challenges in scale-up compared to the other approaches for the treatment of brain diseases, when undesired activity of MMP-9 increases in the brain. However, it should be noted that, we noticed a slow conjugation process for the cholesteryl chloroformate.…”

How scars shape the neural landscape: Key molecular mediators of TGF-β1’s anti-neuritogenic effects

“…Nor do we know if topical applications to the ocular surface can sufficiently target corneal nerve terminals, of whether more sophisticated, cell specific approaches (such as transfection with genetically-encoded materials–e.g. [ 75 , 76 ] are needed to ultimately manipulate GSK-3β activity in damaged, corneal nerves.…”

Biodegradable Nanoparticle for Cornea Drug Delivery: Focus Review

“…The best ratio for achieving the highest transfection efficiency was N:P 30, with no significant cytotoxicity [ 115 ]. Solid based nanoparticle was developed by Torrecilla et al for short-hairpin RNA (shRNA) gene delivery with the main objective of downregulation metalloproteinase 9 (MMP-9) in cornea cells for neovascularization treatment [ 116 ]. Recently, two different types of positive surface charge solid nanoparticles with different ligands (protamine, dextran, or hyaluronic acid (HA)) and synthesized by polyvinyl alcohol (PVA) were developed by Vicente-Pascua et al [ 117 ].…”