Highly versatile cell-penetrating peptide loaded scaffold for efficient and localised gene delivery to multiple cell types: From development to application in tissue engineering

Raftery, Rosanne M.; Walsh, D.; Ferreras, Lia Blokpoel; Castaño, Irene Mencía; Chen, Gang; Lemoine, Mark; Osman, Gizem; Shakesheff, Kevin M.; Dixon, James E.; O’Brien, Fergal J.

doi:10.1016/j.biomaterials.2019.119277

Cited by 58 publications

(88 citation statements)

References 97 publications

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“…GET is suitable for all three germ layer cell transfections with efficiencies comparable to Lipofectamine 3000 and minimal cytotoxicity. These findings suggest that GET can be combined with scaffold delivery systems, to provide new solutions to a variety of tissue engineering regenerative indications (Raftery et al, 2019).…”

Section: Non-viral Gene Delivery Vectorsmentioning

confidence: 95%

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Scaffold-Based Gene Therapeutics for Osteochondral Tissue Engineering

et al. 2020

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Significant progress in osteochondral tissue engineering has been made for biomaterials designed to deliver growth factors that promote tissue regeneration. However, due to diffusion characteristics of hydrogels, the accurate delivery of signaling molecules remains a challenge. In comparison to the direct delivery of growth factors, gene therapy can overcome these challenges by allowing the simultaneous delivery of growth factors and transcription factors, thereby enhancing the multifactorial processes of tissue formation. Scaffold-based gene therapy provides a promising approach for tissue engineering through transfecting cells to enhance the sustained expression of the protein of interest or through silencing target genes associated with bone and joint disease. Reports of the efficacy of gene therapy to regenerate bone/cartilage tissue regeneration are widespread, but reviews on osteochondral tissue engineering using scaffold-based gene therapy are sparse. Herein, we review the recent advances in gene therapy with a focus on tissue engineering scaffolds for osteochondral regeneration.

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Section: Non-viral Gene Delivery Vectorsmentioning

confidence: 95%

“…Compared with protein-based treatment, gene therapy has two main advantages. Gene therapy is more biologically active and physiological than common recombinant approaches (Raftery et al, 2019). Since the gene fragment itself cannot be efficiently introduced into the cell, an effective vector is required.…”

Section: Vector Based Gene-deliverymentioning

confidence: 99%

Scaffold-Based Gene Therapeutics for Osteochondral Tissue Engineering

et al. 2020

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“…Mesenchymal stromal cells (MSCs) are the most employed precursor cell lineage for advanced bone tissue regenerative strategies. Studies which use biomaterials along with exogenous MSCs or those that rely on their endogenous populations have shown significantly enhanced bone formation in critical-size defects when exogenous growth factors (GFs) are applied (Baylink et al, 1993;Kolambkar et al, 2011;Raftery et al, 2019). Numerous GFs have been identified as bone inducer molecules, amongst them bone morphogenetic proteins (BMPs) are the most widely employed GF family with BMP2 and BMP7 being the most prolifically applied FDA approved GFs used clinically for orthopedics (Urist, 1965;Lee, 1997;Groeneveld and Burger, 2000).…”

Section: Introductionmentioning

confidence: 99%

“…Importantly we have shown GET's utility in regenerative medicine by delivering transcription factors RUNX2 and MYOD for osteogenesis and zonal myogenesis in three−dimensional gradients (Eltaher et al, 2016;Thiagarajan et al, 2017), respectively. Moreover, GET peptides have been used to enhance the delivery and transfection of nucleic acids for lung gene therapy and bone regeneration in vivo (Osman et al, 2018;Raftery et al, 2019). The later delivering GF genes to enhance the repair of a critical size calvarial bone defect in rats (Raftery et al, 2019).…”

Section: Introductionmentioning

confidence: 99%

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Efficient Delivery of Transducing Polymer Nanoparticles for Gene-Mediated Induction of Osteogenesis for Bone Regeneration

Jalal

Dixon

2020

Front. Bioeng. Biotechnol.

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Developing non-viral gene therapy vectors that both protect and functionally deliver nucleic acid cargoes will be vital if gene augmentation and editing strategies are to be effectively combined with advanced regenerative medicine approaches. Currently such methodologies utilize high concentrations of recombinant growth factors, which result in toxicity and off-target effects. Herein we demonstrate the use of modified cell penetrating peptides (CPPs), termed Glycosaminoglycan (GAG)-binding Enhanced Transduction (GET) peptides with plasmid DNA (pDNA) encapsulated poly (lacticco-glycolic acid) PLGA nanoparticles (pDNA-encapsulated PLGA NPs). In order to encapsulate the pDNA, it was first condensed with a cationic low molecular weight Poly L-Lysine (PLL) into 30-60 nm NPs followed by encapsulation in PLGA NPs by double emulsion; yielding encapsulation efficiencies (EE) of ∼30%. PLGA NPs complexed with GET peptides show enhanced intracellular delivery (up to sevenfold) and transfection efficiencies (up to five orders of magnitude). Moreover, the pDNA cargo has enhanced protection from nucleases (such as DNase I) promoting their translatability. As an example, we show these NPs efficiently deliver pBMP2 which can promote osteogenic differentiation in vitro. Gene delivery to human Mesenchymal Stromal Cells (hMSCs) inducing their osteogenic programming was confirmed by Alizarin red calcium staining and bone lineage specific gene expression (Q RT-PCR). By combining simplistic and FDA-approved PLGA polymer nanotechnology with the GET delivery system, therapeutic non-viral vectors could have significant impact in future cellular therapy and regenerative medicine applications.

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Enhanced Cellular Transduction of Nanoparticles Resistant to Rapidly Forming Plasma Protein Coronas

et al. 2020

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physical properties suitable for directly targeting therapeutic sites using magnetic fields. [2] Furthermore, the ability of MNPs (a subtype being superparamagnetic iron oxide NPs; SPIONs) to efficiently convert magnetic energy into thermal energy, makes them a focus of interest for use in hyperthermia-based cell ablation anticancer therapies (such as Magforce Nanotechnologies AG). [3-6] Other relevant applications include imaging (including a focus on regenerative medicine), tissue engineering, for mechanical stimulation for cell differentiation in vivo [7-17] and, most recently, their use as synthetic enzymes in biocatalytic processes. [18] Targeted delivery strategies for MNPs have been developed toward specific-or overexpressed receptors on diseased cells by functionalizing the NP surface with proteins, antibodies, or other biomolecules as targeting ligands. These strategies efficiently enhance NP delivery to the target cells in vitro; however, there is mounting evidence that targeting ability of functionalized NPs disappears when placed in an in vivo biological environment. [19-21] It is now well established that when any material surface encounters a biological system, interactions occur between the material and the system components (i.e. proteins, lipids, DNA) forming a layer termed the protein corona. The protein corona defines the biological identity of the particle or surface, and has proven to be an obstacle in the past for effective targeted delivery of ligands, since it affects the physicochemical Nanoparticles (NPs) are increasingly being developed as biomedical platforms for drug/nucleic acid delivery and imaging. However, in biological fluids, NPs interact with a wide range of proteins that form a coating known as protein corona. Coronae can critically influence self-interaction and binding of other molecules, which can affect toxicity, promote cell activation, and inhibit general or specific cellular uptake. Glycosaminoglycan (GAG)-binding enhanced transduction (GET) is developed to efficiently deliver a variety of cargoes intracellularly; employing GAG-binding peptides, which promote cell targeting, and cell penetrating peptides (CPPs) which enhance endocytotic cell internalization. Herein, it is demonstrated that GET peptide coatings can mediate sustained intracellular transduction of magnetic NPs (MNPs), even in the presence of serum or plasma. NP colloidal stability, physicochemical properties, toxicity and cellular uptake are investigated. Using label-free snapshot proteomics, time-resolved profiles of human plasma coronas formed on functionalized GET-MNPs demonstrate that coronae quickly form (<1 min), with their composition relatively stable but evolving. Importantly GET-MNPs present a subtly different corona composition to MNPs alone, consistent with GAG-binding activities. Understanding how NPs interact with biological systems and can retain enhanced intracellular transduction will facilitate novel drug delivery approaches for cell-type specific targeting of new nanomaterials.

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Highly versatile cell-penetrating peptide loaded scaffold for efficient and localised gene delivery to multiple cell types: From development to application in tissue engineering

Cited by 58 publications

References 97 publications

Scaffold-Based Gene Therapeutics for Osteochondral Tissue Engineering

Scaffold-Based Gene Therapeutics for Osteochondral Tissue Engineering

Efficient Delivery of Transducing Polymer Nanoparticles for Gene-Mediated Induction of Osteogenesis for Bone Regeneration

Enhanced Cellular Transduction of Nanoparticles Resistant to Rapidly Forming Plasma Protein Coronas

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