Abstract
The novel synthesis of metal-containing biohybrids using self-assembly methods at physiological temperatures (37 °C) was compared for copper and silver using the amino acid dimer cystine. Once assembled, the copper containing biohybrid is a stable, high-aspect ratio structure, which we call CuHARS. Using the same synthesis conditions, but replacing copper with silver, we have synthesized cystine-capped silver nanoparticles (AgCysNPs), which are shown here to form stable colloid solutions in contrast to the CuHARS, which settle out from a 1 mg/mL solution in 90 min. Both the copper and silver biohybrids, as synthesized, demonstrate very low agglomeration which we have applied for the purpose of applications with cell culture methods, namely, for testing as anti-cancer compounds. AgCysNPs (1000 ng/mL) demonstrated significant toxicity (only 6.8% viability) to glioma and neuroblastoma cells in vitro, with concentrations as low as 20 ng/mL causing some toxicity. In contrast, CuHARS required at least 5 μg/mL. For comparative purposes, silver sulfate at 100 ng/mL decreased viability by 52% and copper sulfate at 100 ng/mL only by 19.5% on glioma cells. Using these methods, the novel materials were tested here as metal–organic biohybrids (MOBs), and it is anticipated that the functionalization and dynamics of MOBs may result in building a foundation of new materials for cellular applications, including cell engineering of both normal and diseased cells and tissue constructs.
Highlights
Bottom-up self-assembly of nanomaterials in general has the advantage of scalability for a given product, with the concept being that once synthesis conditions have been defined, for the case of liquid systems, the volume of the synthesis production may be increased to give a greater yield
As a second contrasting characteristic compared to AgCysNPs, when exposed to two-photon pulses using multi-photon microscopy techniques, the CuHARS demonstrated sufficient second harmonic generation (SHG) such that these metal–organic biohybrids (MOBs) could be illuminated (Figure 7A,B)
We described the characterization and synthesis of self-assembled biohybrids composed of copper and the amino acid dimer cystine [1,2,3]
Summary
Bottom-up self-assembly of nanomaterials in general has the advantage of scalability for a given product, with the concept being that once synthesis conditions have been defined, for the case of liquid systems, the volume of the synthesis production may be increased to give a greater yield. By immobilizing MOBs onto cell culture surfaces in the form of nanofilms, a starting point was established for evaluating the diffusion and breakdown of materials for cell delivery as well as providing an environment for testing cellular outcomes over time This novel approach developed here of layering and drying materials first onto the surface to be tested for toxicity and introducing cells, was compared to the traditional method of plating cells first, followed by addition of nanomaterials on top of them [1,12,13]
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