Water stable, red emitting, carbon nanoparticles stimulate 3D cell invasion via clathrin-mediated endocytic uptake Udisha Singh a , Aditya Guduru Teja b , Shanka Walia a , Payal Vaswani a , Sameer Dalvi bc and Dhiraj Bhatia ac a Biological Engineering Discipline, Indian Institute of Technology Gandhinagar, Palaj, Gujarat 382355, India. E-mail: dhiraj.bhatia@iitgn.ac.in b Chemical Engineering Discipline, Indian Institute of Technology Gandhinagar, Palaj, Gujarat 382355, India c Center for Biomedical Engineering, Indian Institute of Technology Gandhinagar, Palaj, Gujarat 382355, India Bioimaging specific biomolecules or cellular processes or entire cells or tissues is one of the best non-invasive ways to understand and visualise biological activity at the cellular level. Many fluorescent nanomaterials have already been used, such as rare-earth semiconductor quantum dots, organic dyes and polymer dots. However, their applications are still extremely limited due to their limited stability, toxicity, poor water stability and excitation spectra in the UV region, thus hindering their use for biomedical and biological applications(1). Carbon-based nanomaterials, especially carbon dots (CDs, including carbon nanoparticles, graphene quantum dots, and carbon quantum dots), have gained much attention because of their small size, biocompatibility, low toxicity, and stable fluorescent properties, and low cost of synthesis. In the current scenario, we need to synthesise bright fluorescent nanoparticles with excitation and emission towards the red end of the spectrum, which is highly desirable in bioimaging. We synthesised a new class of organic carbon-based nanoparticles (CNPs) with robust quantum yield and fluorescence towards the red region of the spectrum. These are disc-shaped graphene nanoparticles synthesised using organic substrates like para-phenylenediamine (PPDA) dispersed in diphenyl ether and reflux conditions; we achieved scalable amounts of CNPs of the average size of 27 nm and topological heights of 2.6 nm, indicating that fluorescent CNPs contain an average of 7 to 8 layers of graphene sheets. Before using these fluorescent CNPs for biological studies, we check their stability in different solvents, different pH stability, ionic stability, photostability and thermal stability. These CNPs were readily uptaken by different mammalian cells, and we have also studied that they prefer clathrin-mediated endocytosis for their cellular entry route. Not only can these CNPs be specifically uptaken in cells, but they also stimulate cellular processes like cell invasion from 3D spheroid models. These new classes of CNPs, which have sizes similar to proteinaceous ligands, hold immense potential for their surface functionalisation. These could be explored as promising bioimaging agents for biomedical imaging and intracellular drug delivery (2). Keywords: Carbon Nanoparticles, Clathrin-mediated endocytosis, 3D cell invasion, Bioimaging References 1. Wang, Youfu, and Aiguo Hu. "Carbon quantum dots: synthesis, properties and applications." Journal of Materials Chemistry C 2, no. 34 (2014): 6921-6939.Singh, Udisha, Aditya Guduru Teja, Shanka Walia, Payal Vaswani, Sameer Dalvi, and Dhiraj Bhatia. "Water stable, red emitting, carbon nanoparticles stimulate 3D cell invasion via clathrin-mediated endocytic uptake." Nanoscale Advances 4, no. 5 (2022): 1375-1386.
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