Preparation and optimisation of anionic liposomes for delivery of small peptides and cDNA to human corneal epithelial cells

Abstract

Drug delivery to corneal epithelial cells is challenging due to the intrinsic mechanisms that protect the eye. Here we report a novel liposomal formulation to encapsulate and deliver a short sequence peptide into human corneal epithelial cells (hTCEpi). Using a mixture of Phosphatidylcholine/Caproylamine/Dioleoylphosphatidylethanolamine (PC/CAP/DOPE), we encapsulated a fluorescent peptide, resulting in anionic liposomes with an average size of 138.8 ± 34 nm and a charge of −18.2 ± 1.3 mV. After 2 h incubation with the peptide-encapsulated liposomes, 66% of corneal epithelial (hTCEpi) cells internalized the FITC-labelled peptide, demonstrating the ability of this formulation to effectively deliver peptide to hTCEpi cells. Additionally, lipoplexes (liposomes complexed with plasmid DNA) were also able to transfect hTCEpi cells, albeit at a modest level (8% of the cells). Here, we describe this novel anionic liposomal formulation intended to enhance the delivery of small cargo molecules in situ.

Materials promoting viral gene delivery

“…123,422,437 The mechanism that promotes interactions of anionic liposomes with plasma membranes is still not fully understood. Possible reasons, such as longer interaction time due to reduced clearance and toxicity, 437,439,440 faster release of cargo, 441,442 or receptor mediated mechanisms 443 have been discussed. A thorough understanding of mechanisms promoting interactions of anionic liposomes with VPs is yet to be achieved.…”

Transplantation of engineered exosomes derived from bone marrow mesenchymal stromal cells ameliorate diabetic peripheral neuropathy under electrical stimulation

“…Cationic liposomes are associated with certain drawbacks related to in-vitro and in-vivo cytotoxicity and may not be used for the delivery approach [ 37 ]. Therefore, in the present study, anionic liposomes were prepared that may be preferred for the delivery of the nanoparticle's cargo into cells [ 38 ]. TEM images of the synthesized liposomes indicated the near-circular type morphology and nano dimensions ( Fig.…”

“…Anionic liposomes showed slower release kinetics of clearance. They helped to extravagate the cargo inside, better serve our purpose of delivering the cargo effectively and efficiently to the cells for performing therapeutic action [ 38 ]. Anionic liposomes have a better association with the cell membrane that allows its fusion and also overcome the barriers of cytotoxicity and instability associated with cationic liposomes.…”

Biodegradable Nanoparticle for Cornea Drug Delivery: Focus Review

“…Peptides were efficiently delivered to human cornea epithelial cells through liposome-based peptide delivery. Moreover, the use of anionic which was complexed with plasmid DNA culminated in a modest while advantageous transfection of human cornea epithelial cells [ 67 ]. The preparation procedure of these liposomes is schematically shown in Figure 6 .…”