Gene delivery using dendrimer/pDNA complexes immobilized in electrospun fibers using the Layer-by-Layer technique

Ramalingam, Kirthiga; Castro, Rita; Pires, P.F.; Shi, Xiangyang; Rodrigues, João; Xiao, Shili; Tomás, Helena

doi:10.1039/c6ra22444j

Cited by 16 publications

(7 citation statements)

References 106 publications

(87 reference statements)

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“…Sakai et al (2009), por exemplo, modificaram a superfície de fibras eletrofiadas de PLLA com a adsorção camada a camada (LbL) de complexos de PEI e DNA presentes em solução. Ramalingam et al (2016), por sua vez, utilizaram a técnica camada por camada para a formação de nanocamadas de quitosana (polímero catiônico) e alginato (polímero aniônico). Nesse experimento, complexos de pDNA-dendrímeros e pDNA-PEI foram imobilizados nas fibras a partir da interação em uma dispersão.…”

Section: Modificação Superficialunclassified

Complexos de nanofibras eletrofiadas e ácidos nucleicos: Uma revisão

Cavalcante

2021

RSD

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A interação entre células e nanoestruturas poliméricas vem sendo considerado um importante tema para a biotecnologia. As fibras eletrofiadas apresentam estruturas porosas na ordem de submícrons com características que mimetizam os componentes fibrilares da matriz extracelular natural, favorecendo a atuação local efetiva de sistemas bioativos. A complexação de ácidos nucleicos com essas fibras e sua utilização em terapias de recuperação funcional e regeneração tecidual podem ser alternativas ao transplante celular e aos sistemas de entrega de proteínas indutivas. Na customização do perfil de interação entre essas matrizes e o material genético, é relevante manter a disponibilidade de seus tipos moleculares em concentrações efetivas no microambiente, com maior expressão gênica e maior tempo de ação terapêutica. Ao se buscar equilíbrio entre eficiência de transfecção e a viabilidade celular, diferentes estratégias são seguidas, como a incorporação de polinucleotídeos em soluções poliméricas ou em emulsões antes do processo de eletrofiação, ou mesmo a funcionalização de nanofibras pela modificação de sua superfície. Diante disso, esta revisão tem por finalidade apresentar os diferentes métodos de produção de nanofibras eletrofiadas funcionalizadas com ácidos nucleicos e suas aplicações na área da saúde.

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Section: Modificação Superficialunclassified

Complexos de nanofibras eletrofiadas e ácidos nucleicos: Uma revisão

Cavalcante

2021

RSD

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“…The dispersed were subsequently centrifuged at 10,000 rpm at 4 • C for 20 min, and the supernatant liquid containing the released DNA and DNA-LPEI complex was collected to determine the release behavior. A Quant-iT PicoGreen DNA assay kit was used to characterize and quantify the release profile of pDNA from LPEI [52]. After incubating for 1, 2, 4, 7, and 14 days, the collected supernatant was quantified using fluorometry.…”

Section: Loading Confirmation and Release Behavior Studymentioning

confidence: 99%

Fabrication and Biological Activities of Plasmid DNA Gene Carrier Nanoparticles Based on Biodegradable l-Tyrosine Polyurethane

et al. 2021

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Gene therapy is a suitable alternative to chemotherapy due to the complications of drug resistance and toxicity of drugs, and is also known to reduce the occurrence of cellular mutation through the use of gene carriers. In this study, gene carrier nanoparticles with minimal toxicity and high transfection efficiency were fabricated from a biocompatible and biodegradable polymer, l-tyrosine polyurethane (LTU), which was polymerized from presynthesized desaminotyrosyl tyrosine hexyl ester (DTH) and polyethylene glycol (PEG), by using double emulsion and solvent evaporation techniques, resulting in the formation of porous nanoparticles, and then used to evaluate their potential biological activities through molecular controlled release and transfection studies. To assess cellular uptake and transfection efficiency, two model drugs, fluorescently labeled bovine serum albumin (FITC-BSA) and plasmid DNA-linear polyethylenimine (LPEI) complex, were successfully encapsulated in nanoparticles, and their transfection properties and cytotoxicities were evaluated in LX2 as a normal cell and in HepG2 and MCF7 as cancer cells. The morphology and average diameter of the LTU nanoparticles were confirmed using light microscopy, transmission electron microscopy, and dynamic light scattering, while confocal microscopy was used to validate the cellular uptake of FITC-BSA-encapsulated LTU nanoparticles. Moreover, the successful cellular uptake of LTU nanoparticles encapsulated with pDNA-LPEI and the high transfection efficiency, confirmed by gel electrophoresis and X-gal assay transfection, indicated that LTU nanoparticles had excellent cell adsorption ability, facilitated gene encapsulation, and showed the sustained release tendency of genes through transfection experiments, with an optimal concentration ratio of pDNA and LPEI of 1:10. All the above characteristics are ideal for gene carriers designed to transport and release drugs into the cytoplasm, thus facilitating effective gene therapy.

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“…Fabrication of G5•NH 2 Grafted Electrospun PLGA (G5•NH 2g-PLGA) Nanofibrous Mats. Electrospun PLGA nanofibrous mats were first prepared and then coated with polyelectrolyte polymers, namely, PDADMAC and PAA, through layer-by-layer electrostatic assembly technology to form (PAA/PDADMAC)-assembled PLGA nanofibers according to the literature report, 34 which has also been described in detail in Supporting Information (SI). In this work, samples (PAA/PDADMAC) 1 -assembled PLGA and (PAA/PDAD-MAC) 2 -assembled PLGA nanofibrous mats were prepared, named PLGA 1 and PLGA 2 , respectively.…”

Section: ■ Experimental Sectionmentioning

confidence: 99%

“…In a previous work, electrospun PLGA nanofibrous scaffolds were functionalized with PAMAM/pDNA complex for substrate-mediated gene delivery through the layer-by-layer (LbL) self-assembly technology, an attractive technique that uses the alternative deposition of oppositely charged polyelectrolytes onto substrate surface to mediate their surface characteristics . This gene-activated PLGA nanofibrous scaffold not only supported the attachment and growth of hMSCs but also provided a controlled release of pDNA/dendrimer complex and an enhanced cell differentiation toward the osteoblastic lineage when pDNA codifying BMP-2 was used.…”

Section: Introductionmentioning

confidence: 99%

Preparation of [Amine-Terminated Generation 5 Poly(amidoamine)]-graft-Poly(lactic-co-glycolic acid) Electrospun Nanofibrous Mats for Scaffold-Mediated Gene Transfection

Xiao

Peng

Yang

et al. 2019

ACS Appl. Bio Mater.

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Combining biomaterial scaffolds with gene cargos for gene therapy is promising for tissue engineering. Herein, we developed a gene delivery platform through surface grafting of amine-terminated generation 5 poly(amidoamine) (PAMAM) dendrimers (G5·NH2) with biodegradable electrospun poly(lactic-co-glycolic acid) (PLGA) nanofibers by combining layer-by-layer (LbL) electrostatic assembly technology with dendrimer chemistry. PLGA nanofibers were precoated with positively charged poly(diallydimethylammoium chloride) and poly(acrylic acid) through electrostatic interaction and then subsequently cross-linked with G5·NH2 dendrimer covalently through 1-ethyl-3-[3-(dimethylamino)propyl] carbodiimide hydrochloride chemistry. The successful grafting of G5·NH2 dendrimer on PLGA nanofibers was confirmed by X-ray photoelectron spectroscopy. Scanning electron microscopy studies show that smooth, uniform morphology of nanofibers does not significantly change after grafting of G5·NH2 dendrimers except for a slight increase in the fiber diameter, whereas atomic force microscopy images at a high-resolution scale indicated a slightly rough surface for PLGA nanofibers after grafting with G5·NH2 dendrimer. Additionally, PLGA nanofibrous scaffolds became hydrophilic after grafting with G5·NH2 dendrimers. Biological investigation showed that the developed G5·NH2-g-PLGA nanofibrous scaffolds not only allowed for the attachment and proliferation of NIH 3T3 cells but also were capable of complexing pDNA and delivering pDNA/dendrimer complex for solid state gene transfection in situ. The functionalization of PLGA nanofibers with dendrimers may find diverse applications in the area of tissue engineering, gene therapy, and drug delivery.

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Gene delivery using dendrimer/pDNA complexes immobilized in electrospun fibers using the Layer-by-Layer technique

Abstract: Dendrimer/pDNA complexes can be immobilized in PLGA fibers through the Layer-by-Layer technique and direct hMSCs towards osteogenic differentiation.

Cited by 16 publications

References 106 publications

Complexos de nanofibras eletrofiadas e ácidos nucleicos: Uma revisão

Complexos de nanofibras eletrofiadas e ácidos nucleicos: Uma revisão

Fabrication and Biological Activities of Plasmid DNA Gene Carrier Nanoparticles Based on Biodegradable l-Tyrosine Polyurethane

Preparation of [Amine-Terminated Generation 5 Poly(amidoamine)]-graft-Poly(lactic-co-glycolic acid) Electrospun Nanofibrous Mats for Scaffold-Mediated Gene Transfection

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