Spatial control of gene expression within a scaffold by localized inducer release

Baraniak, Priya R.; Nelson, Debra L.; Leeson, Cory E.; Katakam, Anand Kumar; Friz, Jennifer L.; Cress, Dean E.; Hong, Yi; Guan, Jianjun; Wagner, William R.

doi:10.1016/j.biomaterials.2010.12.037

Cited by 17 publications

(21 citation statements)

References 59 publications

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“…The low burst release and the long-term release may be attributed to the hydrogen bonding between DPA and BPU, and the interaction between the hydrophobic DPA and the hydrophobic segment of BPU. The same phenomenon was found from a long-term release of RheoSwitch Ligand 1 molecule (RSL1) loaded in a biodegradable polyurethane scaffold, where the hydrophobic RSL1 interacts with the polyurethane [35]. …”

Section: Discussionmentioning

confidence: 63%

Electrospun biodegradable elastic polyurethane scaffolds with dipyridamole release for small diameter vascular grafts

et al. 2014

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Acellular biodegradable small diameter vascular grafts (SDVGs) require antithrombosis, intimal hyperplasia inhibition and rapid endothelialization to improve the graft patency. However, current antithrombosis and antiproliferation approaches often conflict with endothelial cell layer formation on SDVGs. To address this limitation, biodegradable elastic polyurethane urea (BPU) and the drug dipyridamole (DPA) were mixed and then electrospun into a biodegradable fibrous scaffold. The BPU would provide the appropriate mechanical support, while the DPA in the scaffold would offer biofunctions as required above. We found that the resulting scaffolds had tensile strengths and strains comparable with human coronary artery. The DPA in the scaffolds was continuously released up to 91 days in phosphate buffer solution at 37 °C, with a low burst release within the first 3 days. Compared to BPU alone, improved non-thrombogenicity of the DPA-loaded BPU scaffolds was evidenced with extended human blood clotting time, lower TAT complex concentration, lower hemolysis and reduced human platelet deposition. The scaffolds with a higher DPA content (5 and 10%) inhibited proliferation of human aortic smooth muscle cell significantly. Furthermore, the DPA-loaded scaffolds had no adverse effect on human aortic endothelial cell growth, yet it improved their proliferation. The attractive mechanical properties and biofunctions of the DPA-loaded BPU scaffold indicate its potential as an acellular biodegradable SDVG for vascular replacement.

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Section: Discussionmentioning

confidence: 63%

Electrospun biodegradable elastic polyurethane scaffolds with dipyridamole release for small diameter vascular grafts

et al. 2014

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“…Tight localization of ADV and the resulting spatial control of cell adherence could have utility in in vitro applications of tissue regeneration, for example in the patterning of cell co-cultures. [37,55] …”

Section: Resultsmentioning

confidence: 99%

In Situ Transfection by Controlled Release of Lipoplexes Using Acoustic Droplet Vaporization

Juliar

Moncion

et al. 2016

Adv Healthcare Materials

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Localized delivery of nucleic acids to target sites (e.g., diseased tissue) is critical for safe and efficacious gene therapy. An ultrasound-based technique termed acoustic droplet vaporization (ADV) has been used to spatiotemporally control the release of therapeutic small molecules and proteins contained within sonosensitive emulsions. Here, ADV was used to control the release of lipoplex – containing plasmid DNA with an enhanced green fluorescent protein (eGFP) reporter - from a sonosensitive emulsion. Focused ultrasound (3.5 MHz, mechanical index (MI) ≥ 1.5) generated robust release of fluorescein (i.e., surrogate payload) and lipoplex from the emulsion. In situ release of the lipoplex from the emulsion using ADV (MI=1.5, 30 cycles) yielded a 55% release efficiency, resulting in 43% transfection efficiency and 95% viability with C3H/10T1/2 cells. Without exposure to ultrasound, the release and transfection efficiencies were 5% and 7%, respectively, with 99% viability. Lipoplex released by ADV retained its bioactivity while the ADV process did not yield any measureable sonoporative enhancement of transfection. Co-encapsulation of Ficoll PM 400 within the lipoplex loaded emulsion, and its subsequent release using ADV, yielded higher transfection efficiency than the lipoplex alone. The results demonstrate that ADV could have utility in the spatiotemporal control of gene delivery.

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“…One example is an AND-gate circuit in which gene expression would be turned on or off under specific well-defined biological circumstances. The result could be a genetically interactive matrix in which successive cellular differentiation events could be triggered in a spatial and/or temporal manner (33).…”

Section: Discussionmentioning

confidence: 99%

“…As the PLGA scaffold slowly degrades over time in culture, IPTG is released, allowing cellular uptake and activation of gene expression by the surrounding cells. The passive release of a genetic inducer from porous scaffolds has similarly been used to deliver the inducer, RSL1, from poly(ester urethane) urea (PEUU) films to overlying cells in culture (33). In contrast to RSL1, which is hydrophobic, IPTG is hydrophilic, soluble, and more amenable to delivery in culture and in vivo.…”

Section: Iptg Diffuses Through Various Materials and Activates Genetimentioning

confidence: 99%

“…For in vivo spatial and temporal induction, we sought to investigate whether we could activate and control EGFP expression by administering IPTG to the mice in their drinking water. IPTG is readily soluble in water so systemic administration is more practical experimentally and therapeutically than hydrophobic inducers, such as RSL1 or tamoxifen (33,37). PLGA sponges and PEG hydrogels lacking IPTG but containing CHO cells stably transfected with LTRi_ EGFP were implanted in the abdominal cavity of athymic mice.…”

Section: Iptg Diffuses Through Various Materials and Activates Genetimentioning

confidence: 99%

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Regulating synthetic gene networks in 3D materials

Deans

Singh

Gibson

et al. 2012

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

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Combining synthetic biology and materials science will enable more advanced studies of cellular regulatory processes, in addition to facilitating therapeutic applications of engineered gene networks. One approach is to couple genetic inducers into biomaterials, thereby generating 3D microenvironments that are capable of controlling intrinsic and extrinsic cellular events. Here, we have engineered biomaterials to present the genetic inducer, IPTG, with different modes of activating genetic circuits in vitro and in vivo. Gene circuits were activated in materials with IPTG embedded within the scaffold walls or chemically linked to the matrix. In addition, systemic applications of IPTG were used to induce genetic circuits in cells encapsulated into materials and implanted in vivo. The flexibility of modifying biomaterials with genetic inducers allows for patterned placement of these inducers that can be used to generate distinct patterns of gene expression. Together, these genetically interactive materials can be used to characterize genetic circuits in environments that more closely mimic cells' natural 3D settings, to better explore complex cell-matrix and cell-cell interactions, and to facilitate therapeutic applications of synthetic biology.T he complexity of cell signaling can be simplified by considering genetic networks composed of subsets of simpler parts, or modules. This simplification is the foundation of synthetic biology, where engineering paradigms are applied in rational and systematic ways to produce predictable and robust systems for understanding mechanisms of cellular function (1-6). The majority of work in synthetic biology has been in simple organisms, such as yeast and bacteria. However, as synthetic biology starts to expand to mammalian systems, it becomes increasingly more important to consider the environment in which the cells are grown. Biomaterials will play an important role in advancing synthetic biology within mammalian systems, because they provide highly controllable and tunable microenvironments where cells can behave as they do in vivo, in addition to organizing and delivering therapeutic cells to locations of interest. Our results show that interfacing synthetic biology and biomaterials can catalyze synthetic biology applications through engineered biomaterials that actively control genetic circuits in 3D scaffolds to more closely mimic the cells' natural settings, in addition to providing mechanisms for translating synthetic biology for clinical applications (Fig. S1).Biomaterials provide 3D environments for cell growth and have rapidly advanced our ability to investigate the coordinated interactions of many cellular phenomena because biomaterials recapitulate the in vivo setting better than traditional 2D cultures, where cells are grown in monolayers (7,8). Physiological development, homeostasis, and regeneration each require a complex interplay of multiple signals that originate from the extracellular matrix (ECM) and from the intrinsic cellular control of gene products (9). Wh...

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Spatial control of gene expression within a scaffold by localized inducer release

Cited by 17 publications

References 59 publications

Electrospun biodegradable elastic polyurethane scaffolds with dipyridamole release for small diameter vascular grafts

Electrospun biodegradable elastic polyurethane scaffolds with dipyridamole release for small diameter vascular grafts

In Situ Transfection by Controlled Release of Lipoplexes Using Acoustic Droplet Vaporization

Regulating synthetic gene networks in 3D materials

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