Abstract
The anti-inflammatory cytokine Interleukin-10 (IL-10) is considered an efficient treatment for corneal inflammation, in spite of its short half-life and poor eye bioavailability. In the present work, mRNA-based nanomedicinal products based on solid lipid nanoparticles (SLNs) were developed in order to produce IL-10 to treat corneal inflammation. mRNA encoding green fluorescent protein (GFP) or human IL-10 was complexed with different SLNs and ligands. After, physicochemical characterization, transfection efficacy, intracellular disposition, cellular uptake and IL-10 expression of the nanosystems were evaluated in vitro in human corneal epithelial (HCE-2) cells. Energy-dependent mechanisms favoured HCE-2 transfection, whereas protein production was influenced by energy-independent uptake mechanisms. Nanovectors with a mean particle size between 94 and 348 nm and a positive superficial charge were formulated as eye drops containing 1% (w/v) of polyvinyl alcohol (PVA) with 7.1–7.5 pH. After three days of topical administration to mice, all formulations produced GFP in the corneal epithelium of mice. SLNs allowed the obtaining of a higher transfection efficiency than naked mRNA. All formulations produce IL-10, and the interleukin was even observed in the deeper layers of the epithelium of mice depending on the formulation. This work shows the potential application of mRNA-SLN-based nanosystems to address corneal inflammation by gene augmentation therapy.
Highlights
The eye is considered the perfect target for gene therapy due to its physiological features: it is easy to access and examine, it has a well-defined anatomy and it is relatively immune privileged
We showed that solid lipid nanoparticles (SLNs)-based systems bearing pDNA were able to produce up to 10 ng/mL of IL-10 in HCE-2 cells [8]
The nature of the nucleic acid and the target cell are key points for the design and optimization of nanosystems aimed at ocular gene delivery, with mRNA being more sensitive to the formulation-related factors than pDNA
Summary
The eye is considered the perfect target for gene therapy due to its physiological features: it is easy to access and examine, it has a well-defined anatomy and it is relatively immune privileged. 47 clinical trials restricted to “ocular diseases” are registered in the database of Gene Therapy Clinical Trials Worldwide [4], all of them targeting retinal degenerative diseases, except one, which concerns corneal opacity. Several factors can damage this tissue and provoke corneal inflammation or keratitis, such as infections, dry eye, eyelid. Pharmaceutics 2021, 13, 1472 disorders, physical and chemical damage, and a wide variety of underlying diseases [5]. Current treatments of corneal inflammation are based on corticosteroids, which require repeated topical applications and have been associated with multiple adverse effects, such as infectious keratitis, increased intraocular pressure, and cataracts [6]. IL-10 ocular delivery may be better attempted by gene therapy, with the aim of inducing IL-10 de novo synthesis in corneal cells [8]
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