Programmable soft DNA hydrogels stimulate cellular endocytic pathways and proliferation

Ankur Singh; Nihal Singh; Manasi Esther Jinugu; Prachi Thareja; Dhiraj Bhatia

doi:10.1016/j.bioadv.2024.214040

Programmable soft DNA hydrogels stimulate cellular endocytic pathways and proliferation

Biomater Adv. 2024 Sep 12:166:214040. doi: 10.1016/j.bioadv.2024.214040. Online ahead of print.

Authors

Ankur Singh¹, Nihal Singh¹, Manasi Esther Jinugu², Prachi Thareja³, Dhiraj Bhatia⁴

Affiliations

¹ Department of Biological Sciences & Engineering, Indian Institute of Technology Gandhinagar, Palaj, Gujarat 382355, India.
² Department of Chemical Engineering, Indian Institute of Technology Gandhinagar, Palaj, Gujarat 382355, India.
³ Department of Chemical Engineering, Indian Institute of Technology Gandhinagar, Palaj, Gujarat 382355, India; Dr. Kiran C Patel Center for Sustainable Development (KPCSD), Indian Institute of Technology Gandhinagar, India.
⁴ Department of Biological Sciences & Engineering, Indian Institute of Technology Gandhinagar, Palaj, Gujarat 382355, India. Electronic address: dhiraj.bhatia@iitgn.ac.in.

PMID: 39293253
DOI: 10.1016/j.bioadv.2024.214040

Abstract

Hydrogels are pivotal in tissue engineering, regenerative medicine, and drug delivery applications. Existing hydrogel platforms are not easily customizable and often lack precise programmability, making them less suited for 3D tissue culture and programming of cells. DNA molecules stand out among other promising biomaterials due to their unparalleled precision, programmability, and customization. In this study, we introduced a palette of novel cellular scaffolding platforms made of pure DNA-based hydrogel systems while improving the shortcomings of the existing platforms. We showed a quick and easy one step synthesis of DNA hydrogels using thermal annealing based on sequence specific hybridization strategy. We also demonstrated the formation of multi-armed branched supramolecular scaffolds with custom mechanical stiffness, porosity, and network density by increasing or decreasing the number of branching arms. These mechanically tuneable DNA hydrogels proved to be a suitable suitable platform for modulating the physiological processes of retinal pigment epithelial cells (RPE1). In-vitro studies showed dynamic changes at multiple levels, ranging from a change in morphology to protein expression patterns, enhanced membrane traffic, and proliferation. The soft DNA hydrogels explored here are mechanically compliant and pliable, thus excellently suited for applications in cellular programming and reprogramming. This research lays the groundwork for developing a DNA hydrogel system with a higher dynamic range of stiffness, which will open exciting avenues for tissue engineering and beyond.

Keywords: Actin polymerization; Cell proliferation; Cell spreading; DNA hydrogel; Mitochondrial fragmentation; Self-assembled supramolecular frameworks.