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Synthesis, characterization, and functionalization of Graphene Oxide-based nanoplatforms for gene delivery
1 , 2 , * 1
1  Department of Chemical Engineering, Universidad de Los Andes, Bogota, Colombia
2  Department of Biomedical Engineering, Universidad de Los Andes, Bogota, Colombia


Gene therapy has been considered as a promising strategy for the treatment of several inherited disesases and acquired complex disorders . One important challenge yet to be solved to ensure the success of the nanomaterials in the delivery of gene therapies is their ability to escape from endosomes. To address this issue, we previously developed magnetite nanoparticles conjugated with the antimicrobial peptide Buforin II, which showed potent translocating and endosomal escape abilities in several cell lines. In this work, we propose the development of new cell-penetrating nanoplatforms by interfacing graphene oxide (GO) with potent translocating peptides to take advantage of already tested and new peptides as well as the distinctive interactions of GO with the phospholipids of membranes and endosomes. GO was prepared by the modified Hummers’ method through the oxidation of graphite sheets. Next, functionalization of GO was carried out by mixing tetramethylammonium hydroxide (TMAH) 25% (v/v), pure acetic acid and (3-Aminopropyl) triethoxysilane (APTES) 10% (v/v). Thermogravimetric analysis (TGA) and Fourier-Transform Infrared spectroscopy were used to characterize the nanoplatform. FTIR analysis exhibited the peaks related to the characteristic carboxyl groups of GO as well as the Si-O bonds after silanization. TGA allowed us to estimate a silanization efficiency of about 35%. Future work will be focused on conjugating Buforin II and assessing translocation efficiency by conducting uptake assays in liposomes and various cell lines. Additionally, endosomal escape will be determined via confocal microscopy by labeling the peptide with fluorescent molecules and looking at colocalization with the fluorescent probe lysotracker. By taking advantage of the exceptional qualities in terms of physicochemical, electrical and optical properties of GO, this study might provide novel strategies to overcome limitations commonly faced such as low stability of the translocating biomolecules and endosomal entrapment.

Keywords: Gene therapy; Graphene Oxide; functionalization
Comments on this paper
Andreza Lima
Congratulations on your manuscript. How to ensure that the application of graphene for gene delivery will be safe? Do you intend to analyze biocompatibility and toxicity of functionalized graphene oxide?
Julian Torres-Vanegas
I do appreciate your kind comment. Also, I'm glad you ask about biocompability and toxicity of functionalized graphene oxide. Definitely, we have to conduct hemolysis, platelet aggregation and cytotoxicity assays to ensure that our graphene oxide-based nanoplatforms are safe for gene delivery and that's part of the work we will be carrying out in the near future.