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WIREs Nanomed Nanobiotechnol
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From infection to healing: The use of plant viruses in bioactive hydrogels

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Abstract Plant viruses show great diversity in shape and size, but each species forms unique nucleoprotein particles that are symmetrical and monodisperse. The genetically programed structure of plant viruses allows them to be modified by genetic engineering, bioconjugation, or encapsulation to form virus nanoparticles (VNPs) that are suitable for a broad range of applications. Plant VNPs can be used to present foreign proteins or epitopes, to construct inorganic hybrid materials, or to carry molecular cargos, allowing their utilization as imaging reagents, immunomodulators, therapeutics, nanoreactors, and biosensors. The medical applications of plant viruses benefit from their inability to infect and replicate in human cells. The structural properties of plant viruses also make them useful as components of hydrogels for tissue engineering. Hydrogels are three‐dimensional networks composed of hydrophilic polymers that can absorb large amounts of water. They are used as supports for tissue regeneration, as reservoirs for controlled drug release, and are found in contact lenses, many wound healing materials, and hygiene products. They are also useful in ecological applications such as wastewater treatment. Hydrogel‐based matrices are structurally similar to the native extracellular matrix (ECM) and provide a scaffold for the attachment of cells. To fully replicate the functions of the ECM it is necessary to augment hydrogels with biological cues that regulate cellular interactions. This can be achieved by incorporating functionalized VNPs displaying ligands that influence the mechanical characteristics of hydrogels and their biological properties, promoting the survival, proliferation, migration, and differentiation of embedded cells. This article is categorized under: Implantable Materials and Surgical Technologies > Nanomaterials and Implants Biology‐Inspired Nanomaterials > Protein and Virus‐Based Structures Implantable Materials and Surgical Technologies > Nanotechnology in Tissue Repair and Replacement
Modifications and applications of plant virus nanoparticles (VNPs) and virus like particles (VLPs). Plant virus capsids can be modified by genetic engineering, bioconjugation, biomineralization or used for the encapsulation of small molecules. These modifications can be used for applications such as biomaterial design, molecular imaging, and drug delivery
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Plant virus functionalization can modify cell behavior on 2D and 3D scaffolds. (a) Production of TMV‐PEG‐DA hydrogels. (i) Schematic representation of the coupling of PEG‐DA to TMV1cys via a thiol‐Michael reaction resulting in TMV particles functionalized with a double bond (TMV‐PEG‐DA). (ii) Subsequent photo‐polymerization of the attached acrylate functionalities with more PEG‐DA to hydrogels (Reprinted with permission from Southan et al. (2018). Copyright 2018 The Royal Society of Chemistry (RSC) on behalf of the European Society for Photobiology, the European Photochemistry Association, and RSC). (b) Hydrogel films and fibers support neurite outgrowth. (i) Production of 2D‐TMV/PANi/PSS films to support neurite growth. (ii) AFM images of aligned TMV/PANi/PSS nanofibers in capillaries coated with PDDA/PSS2.5 (white arrow shows the direction of the flow; scale bar = 1 μm). (iii) Representative fluorescence (FDA staining) images of PC12 cells cultured on aligned TMV/PANi/PSS for 3 days (Reprinted with permission from Wu, Feng, Zan, Lin, and Wang (2015). Copyright (2015) American Chemical Society). (c) Porous alginate hydrogels can support bone regeneration. (i) Generation of virus‐functionalized porous composite hydrogels. (ii) TMV was added 5 min before the foamy mixture was frozen and lyophilized and the lyophilized sample was cross‐linked with CaCl2. (iii) Scanning electron microscopy (SEM) images of hydrogels and TMV particles, TMV‐PAH at different magnifications. (iv) SEM images of a single cell inside TMV‐PAH, scale bar = 10 μm (Reprinted with permission from Luckanagul et al. (2012). Copyright (2012) American Chemical Society). (d) PVX functionalized with biomimetic cues for the functionalization of agarose hydrogels. (i) Schematic representation of genetically engineered PVX for the presentation of MIP and RGD and a combination thereof. (ii) SEM image of hMSC pseudopodia interconnected with PVX‐RGD‐MIP on TCPS coated with 0.05 mg/cm2 PVX after 8 hr (scale bar = 2 μm). (iii) Human MSCs embedded in 0.5% agarose with 0.2% PVX‐RGD incubated for 14 days, scale bar = 100 μm (Reprinted with permission from Lauria et al. (2017). Copyright 2017 Elsevier)
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The favorable traits of hydrogels for tissue engineering applications
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Schematic summary of the production options for virus nanoparticles (VNPs)/virus like particles (VLPs). Plant VNPs can be produced by molecular farming in plants following inoculation with intact particles, genomic RNA or cDNA, or infection with Agrobacterium tumefaciens carrying a cDNA copy of the virus genome. After infection, plant material is harvested and the tissue is disrupted. Cell debris is removed and the extract is used for particle purification by precipitation and gradient separation. Plant VLPs can be produced by expressing coat proteins in suitable hosts such as plants, bacteria or yeast. After cell lysis, subunits can be isolated by precipitation and chromatography. The particles are assembled in vitro and can be purified as described above for VNPs. Alternatively, VLPs can be produced directly from VNPs by partial or complete disassembly followed by the degradation or capture of the nucleic acids and subsequent capsid reassembly
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Implantable Materials and Surgical Technologies > Nanotechnology in Tissue Repair and Replacement
Biology-Inspired Nanomaterials > Protein and Virus-Based Structures
Implantable Materials and Surgical Technologies > Nanomaterials and Implants

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