Abstract
Gelatin is a biopolymer widely used to synthesize hydrogels for biomedical applications, such as tissue engineering and bioinks for 3D bioprinting. However, as with other biopolymer-based hydrogels, gelatin-hydrogels do not allow precise temporal control of the biomolecule distribution to mimic biological signals involved in biological mechanisms. Leveraging DNA nanotechnology tools to develop a responsive controlled release system via strand displacement has demonstrated the ability to encode logic process, which would enable a more sophisticated design for controlled release. However, this unique and dynamic system has not yet been incorporated within any hydrogels to create a complete release circuit mechanism that closely resembles the sequential distribution of biomolecules observed in the native environment. Here, we designed and synthesized versatile multi-arm DNA motifs that can be easily conjugated within a gelatin hydrogel via click chemistry to incorporate a strand displacement circuit. After validating the incorporation and showing the increased stability of DNA motifs against degradation once embedded in the hydrogel, we demonstrated the ability of our system to release multiple model cargos with temporal specificity by the addition of the trigger strands specific to each cargo. Additionally, we were able to modulate the rate and quantity of cargo release by tuning the sequence of the trigger strands.
Highlights
Hydrogels are three-dimensional (3D) hydrophilic networks of crosslinked polymers that can be designed to mimic the structural, biochemical, and mechanical properties of the extracellular matrix (ECM)
We expected the measured size of 3-way junction (3WJ) and 4WJ from Dynamic Light Scattering (DLS) to be similar since the instrument measures the hydrodynamic radius, and we assumed that the nanostructures are spherical and the arm lengths of the two structures are identical
Our results showed that the release rate was much slower in the hydrogel in comparison with the release observed in solution, which is likely due to the slower diffusion rate of the trigger within the hydrogel
Summary
Hydrogels are three-dimensional (3D) hydrophilic networks of crosslinked polymers that can be designed to mimic the structural, biochemical, and mechanical properties of the extracellular matrix (ECM). We were able to demonstrate the capability to tune the quantity and the rate of cargo release with our SDR system by modifying our trigger strand sequences with the introduction of precisely located mismatches. This further expanded the versatility of the system we developed. The quencher and fluorophore are used to quantify the cargo release in our system
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