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The success of gene therapy is largely dependent on the development of a gene carrier. Recently cell-penetrating peptides (CPPs) have been employed for enhancing the gene and drug delivery efficacy of nano-particles. The feasibility of octaarginine (R8) functionalized graphene oxide (GO) as a novel nano-carrier for gene delivery is investigated. A DNA plasmid expressing enhanced green fluorescent protein (pEGFP) is used as a model gene to study the R8-GO transfection ability into mammalian cells. R8 peptide is conjugated in different ratios (0.1-1.5 μmol per mg of GO) to carboxylated graphene oxide by a two step amidation process. The process of peptide conjugation is analyzed by Fourier transform infrared (FTIR), atomic force microscopy (AFM), UV-vis spectroscopy and X-ray diffraction (XRD). In order to obtain the highest transfection of pEGFP into the cells, the amount of peptide bound to GO is optimized which is evidenced by dynamic light scattering (DLS), zeta potential, TNBS and gel retardation assays. The cytotoxicity of R8-functionalized GO is also tested by MTT assay. The results confirm the successful attachment of R8 peptide to GO. The AFM and XRD results show a significant increase in the thickness of nano graphene oxide sheets (NGOS) from 0.8 to 2-7 nm as well as an increase in the GO interlying space after the R8-functionalization process. A reduction in nano-carrier stability in both aqueous solution and cell culture media is observed when the amount of peptide is increased to more than 1 μmol mg(-1). Gel electrophoresis analysis shows the highest DNA loading on the peptide functionalized GO at the ratios of 0.5 and 1 μmol mg(-1). As a result, the conjugated peptide sample with a peptide molar ratio of 1 μmol per mg of GO shows the highest conjugational efficiency and EGFP gene expression along with improved dispersibility and biocompatibility. Overall, the findings reveal the importance of peptide density on the surface of NGOS in order to obtain the most efficient cell transfection. It is concluded that the R8-conjugated GO could be a promising nano-carrier for gene delivery with relevance in biotechnology therapeutics and clinical applications.

The successful application of nucleic acid-based therapy for the treatment of various cancers is largely dependent on a safe and efficient delivery system. A dual-functionalized graphene oxide (GO)-based nanocarrier with the conjugation of aminated-polyethylene glycol (PEG-diamine) and octa-arginine (R8) for the intracellular delivery of nucleic acids is proposed. The functionalized sites are covalently co-conjugated and the PEG : R8 molar ratio is optimized at 10 : 1 to achieve a hydrocolloidally stable size of 252 ± 2.0 nm with an effective charge of +40.97 ± 1.05 and an amine-rich content of 10.87 ± 0.4 μmol g-1. The uptake of the nanocarrier in breast cancer cell lines, MCF-7 and MDA-MB 231, is investigated. The siRNA and pDNA condensation ability in the presence and absence of enzymes and the endosomal buffering capacity, as well as the intracellular localization of the gene/nanocarrier complex are also evaluated. Furthermore, the delivery of functional genes associated with the nanocarrier is assessed using c-Myc protein knockdown and EGFP gene expression. The effective uptake of the nanocarrier by the cells shows superior cytocompatibility, and protects the siRNA and pDNA against enzyme degradation while inhibiting their migration with N : P ratios of 10 and 5, respectively. The co-conjugation of PEG-diamine and the cationic cell-penetrating peptide (CPP) into the GO nanocarrier also provides a superior internalization efficacy of 85% in comparison with a commercially available transfection reagent. The c-Myc protein knockdown and EGFP expression, which are induced by the nanocarrier, confirm that the optimized PEG-diamine/R8-functionalized GO could effectively deliver pDNA and siRNA into the cells and interfere with gene expression.

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Nowadays, engineering‐based cardiac patches aim to accelerate cardiac regeneration in myocardial infarcted tissues. Considering the fundamental role of cardiac electrophysiology in myocardial function, this study aims to investigate graphene oxide (GO) incorporation in the polyethylene terephthalate (PET) nanofibrous scaffold, as a conductive cardiac patch. The PET/GO nanocomposites are prepared using the uniaxial nozzle and coaxial nozzle electrospinning processes and comprehensively evaluated. The morphological observation indicates a uniform beaded free morphology with an average diameter of 147 ± 38 and 253 ± 67 nm for solid and core–shell nanocomposite fibers, respectively. Addition of GO to the PET nanofibers in a concentration of 0.05 wt% remarkably increases the Young modulus of mats from 30 ± 0.03 to 60 ± 0.02 and 69 ± 0.08 MPa for solid and core–shell nanofibers, respectively. Also, the electroconductivity is improved from 0.7 × 10−6 to 1.175 × 10−6 and 1.3 × 10−6 S cm−1 for solid and core–shell nanofibers, which are in the range of cardiac electroactivity values. PET/GO substrate interestingly supports human umbilical vein endothelial cells’ spreading morphology and cardiomyocyte elongated morphology, mainly where the GO nanosheets are distributed near the surface of nanofibers. In conclusion, the core–shell electrospun PET/GO nanocomposite fibers are suggested as a potential electroactive cardiac patch to improve cardiac cell attachment and proliferation.

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