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Abstract
Conjugation of latent growth factors to superparamagnetic iron oxide nanoparticles (SPIONs) is potentially useful for magnetically triggered release of bioactive macromolecules. Thus, the goal of this work was to trigger the release of active Transforming Growth-Factor Beta (TGF-β) via magnetic hyperthermia by binding SPIONs to the latent form of TGF-β, since heat has been shown to induce release of TGF-β from the latent complex. Commercially available SPIONS with high specific absorption rates (SAR) were hydrolyzed in 70% ethanol to create surface carboxylic acid conjugation sites for carbodiimide chemistry. Fourier-Transform Infra-Red (FTIR) analysis verified the conversion of maleic anhydride to maleic acid. 1-Ethyl-2-(3-dimethyulaminopropyl) carbodiimide (EDC) and N-hydroxysulfosuccinimide (Sulfo-NHS) were used to bind to the open conjugation sites of the SPION in order to graft latent TGF-β onto the particles. The resulting conjugated particles were imaged with transmission electron microscopy (TEM), and the complexed particles were characterized by dynamic light scattering (DLS) and superconducting quantum interference device (SQUID) magnetometry. Enzyme-linked immunosorbent assay (ELISA) was used to assess the thermally triggered release of active TGF-β from the latent complex, demonstrating that conjugation did not interfere with release. Results showed that latent TGF-β was successfully conjugated to the iron oxide nanoparticles, and magnetically triggered release of active TGF-β was achieved.
Highlights
Transforming growth factor beta (TGF-β) is a highly studied cytokine that participates in or controls a wide variety of cell signaling pathways
TGF-β exists sequestered by a latency associated peptide (LAP), forming what is known as the small latent complex (SLC) [13]
This current study aims to replicate the previous work of conjugating latent TGF-β to SPIONS for magneto-thermal activation via the carbodiimide crosslinking chemistry used to conjugate the same protein with carbon nanotubes for near infra-red (NIR) light triggered release
Summary
Transforming growth factor beta (TGF-β) is a highly studied cytokine that participates in or controls a wide variety of cell signaling pathways. These include control of cell cycle [1], cell proliferation [2,3], extracellular matrix formation [4,5], and even stem cell differentiation [6,7]. The active form of TGF-β can be released from the SLC via a variety of different methods, including mechanical unfolding of the LAP by cell surface integrins, as well as chemical interaction with thrombospndin-1 or reactive oxygen species [14,15]. Another method of triggered release combined with greater penetration depth is needed to provide therapeutic effects to deep tissues
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