Abstract
Halloysite nanotubes (HNTs) are natural aluminosilicate clay that have been extensivelyexplored fordelivery of bioactive agents in biomedical applications because of their desirable features including unique hollow tubular structure, good biocompatibility, high mechanical strength, and extensive functionality. For the first time, in this work, functionalized HNTs are developed as a delivery platform for nitric oxide (NO), a gaseous molecule, known for its important roles in the regulation of various physiological processes. HNTs were first hydroxylated and modified with an aminosilane crosslinker, (3-aminopropyl) trimethoxysilane (APTMS), to enable the covalent attachment of a NO donor precursor, N-acetyl-d-penicillamine (NAP). HNT-NAP particles were then converted to NO-releasing S-nitroso-N-acetyl-penicillamine HNT-SNAP by nitrosation. The total NO loading on the resulting nanotubes was 0.10 ± 0.07 μmol/mg which could be released using different stimuli such as heat and light. Qualitative (Fourier-transform infrared spectroscopy and Nuclear magnetic resonance) and quantitative (Ninhydrin and Ellman) analyses were performed to confirm successful functionalization of HNTs at each step. Field emission scanning electron microscopy (FE-SEM) showed that the hollow tubular morphology of the HNTs was preserved after modification. HNT-SNAP showed concentration-dependent antibacterial effects against Gram-positive Staphylococcus aureus (S. aureus), resulting in up to 99.6% killing efficiency at a concentration of 10 mg/mL as compared to the control. Moreover, no significant cytotoxicity toward 3T3 mouse fibroblast cells was observed at concentrations equal or below 2 mg/mL of HNT-SNAP according to a WST-8-based cytotoxicity assay. The SNAP-functionalized HNTs represent a novel and efficient NO delivery system that holds the potential to be used, either alone or in combination with polymers for different biomedical applications.