Cell-controlled and spatially arrayed gene delivery from fibrin hydrogels

Lei, Pedro; Padmashali, Roshan M.; Andreadis, Stelios T.

doi:10.1016/j.biomaterials.2009.03.049

Cited by 93 publications

(89 citation statements)

References 54 publications

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“…Our findings extend previous work showing biomaterialmediated gene delivery using viral vectors (20,(40)(41)(42)(43)(44)(45)(46)(47)(48)(49). The majority of this work demonstrated the ability to specify the spatial location of delivered vectors, rather than the induction of tissuespecific differentiation.…”

Section: Discussionsupporting

confidence: 86%

Scaffold-mediated lentiviral transduction for functional tissue engineering of cartilage

Brunger

Huynh

Guenther

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

116

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The ability to develop tissue constructs with matrix composition and biomechanical properties that promote rapid tissue repair or regeneration remains an enduring challenge in musculoskeletal engineering. Current approaches require extensive cell manipulation ex vivo, using exogenous growth factors to drive tissue-specific differentiation, matrix accumulation, and mechanical properties, thus limiting their potential clinical utility. The ability to induce and maintain differentiation of stem cells in situ could bypass these steps and enhance the success of engineering approaches for tissue regeneration. The goal of this study was to generate a self-contained bioactive scaffold capable of mediating stem cell differentiation and formation of a cartilaginous extracellular matrix (ECM) using a lentivirus-based method. We first showed that poly-L-lysine could immobilize lentivirus to poly(e-caprolactone) films and facilitate human mesenchymal stem cell (hMSC) transduction. We then demonstrated that scaffoldmediated gene delivery of transforming growth factor β3 (TGF-β3), using a 3D woven poly(e-caprolactone) scaffold, induced robust cartilaginous ECM formation by hMSCs. Chondrogenesis induced by scaffold-mediated gene delivery was as effective as traditional differentiation protocols involving medium supplementation with TGF-β3, as assessed by gene expression, biochemical, and biomechanical analyses. Using lentiviral vectors immobilized on a biomechanically functional scaffold, we have developed a system to achieve sustained transgene expression and ECM formation by hMSCs. This method opens new avenues in the development of bioactive implants that circumvent the need for ex vivo tissue generation by enabling the long-term goal of in situ tissue engineering.biomaterials | regenerative medicine | genetic engineering | gene therapy | chondrocyte

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Section: Discussionsupporting

confidence: 86%

Scaffold-mediated lentiviral transduction for functional tissue engineering of cartilage

Brunger

Huynh

Guenther

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

116

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“…Plasmid DNA [42][43][44][45] and recombinant viruses 46,47 have also been incorporated into fibrin hydrogels by our group and others, and they may be also used in this setting to promote vascularization. For the most part, genetic modification in this setting is local to the cells that populate the dermis during the granulation tissue phase of wound healing, for example, macrophages, fibroblasts, and endothelial cells.…”

Section: Discussionmentioning

confidence: 99%

Vascularization of the Dermal Support Enhances Wound Re-Epithelialization by In Situ Delivery of Epidermal Keratinocytes

Lugo

Lei

Andreadis

2011

Tissue Engineering Part A

Self Cite

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Despite significant advances in management of severe wounds such as burns and chronic ulcers, autologous split-thickness skin grafts are still the gold standard of care. The main problems with this approach include pain and discomfort associated with harvesting autologous tissue, limited availability of donor sites, and the need for multiple surgeries. Although tissue engineering has great potential to provide alternative approaches for tissue regeneration, several problems have hampered progress in translating technological advances to clinical reality. Specifically, engineering of skin substitutes requires long culture times and delayed vascularization after implantation compromises graft survival. To address these issues we developed a novel two-prong strategy for tissue regeneration in vivo: (1) vascularization of acellular dermal scaffolds by infiltration of angiogenic factors; and (2) generation of stratified epidermis by in situ delivery of epidermal keratinocytes onto the prevascularized dermal support. Using athymic mouse as a model system, we found that incorporation of angiogenic factors within acellular human dermis enhanced the density and diameter of infiltrating host blood vessels. Increased vascularization correlated with enhanced proliferation and stratification of the neoepidermis originating from the fibrin-keratinocyte cell suspension. This strategy promoted tissue regeneration in vivo with no need for engineering skin substitutes; therefore, it may be useful for treatment of major wounds when skin donor sites are scarce and rapid wound coverage is required.

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“…Although naked DNA has shown gene expression and ability to guide regeneration in vivo (18,24), limitations with low gene transfer efficiency and rapid diffusion of the DNA from the hydrogel scaffold motivated the use of DNA nanoparticles instead of naked DNA. DNA condensed either with cationic peptides, lipids, or polymers has been introduced into fibrin hydrogels (25)(26)(27)(28)(29), enzymatically degradable PEG hydrogels (30), and PEG-hyaluronic acid hydrogels (26). The delivery of genes from hydrogel scaffolds is becoming an attractive route to introduce foreign genes to cells for several reasons.…”

Section: Introductionmentioning

confidence: 99%

Surface- and Hydrogel-Mediated Delivery of Nucleic Acid Nanoparticles

Pannier

Segura

2012

Nanotechnology for Nucleic Acid Delivery

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Gene expression within a cell population can be directly altered through gene delivery approaches. Traditionally for nonviral delivery, plasmids or siRNA molecules, encoding or targeting the gene of interest, are packaged within nanoparticles. These nanoparticles are then delivered to the media surrounding cells seeded onto tissue culture plastic; this technique is termed bolus delivery. Although bolus delivery is widely utilized to screen for efficient delivery vehicles and to study gene function in vitro, this delivery strategy may not result in efficient gene transfer for all cell types or may not identify those delivery vehicles that will be efficient in vivo. Furthermore, bolus delivery cannot be used in applications where patterning of gene expression is needed. In this chapter, we describe methods that incorporate material surfaces (i.e., surface-mediated delivery) or hydrogel scaffolds (i.e., hydrogel-mediated delivery) to efficiently deliver genes. This chapter includes protocols for surface-mediated DNA delivery focusing on the simplest and most effective methods, which include nonspecific immobilization of DNA complexes (both polymer and lipid vectors) onto serum-coated cell culture polystyrene and self-assembled monolayers of alkanethiols on gold. Also, protocols for the encapsulation of DNA/cationic polymer nanoparticles into hydrogel scaffolds are described, including methods for the encapsulation of low amounts of DNA (<0.2 μg/μL) and high amounts of DNA (>0.2 μg/μL) since incorporation of high amounts of DNA poses significant challenges due to aggregation.

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Cell-controlled and spatially arrayed gene delivery from fibrin hydrogels

Cited by 93 publications

References 54 publications

Scaffold-mediated lentiviral transduction for functional tissue engineering of cartilage

Scaffold-mediated lentiviral transduction for functional tissue engineering of cartilage

Vascularization of the Dermal Support Enhances Wound Re-Epithelialization by In Situ Delivery of Epidermal Keratinocytes

Surface- and Hydrogel-Mediated Delivery of Nucleic Acid Nanoparticles

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