Lignin-facilitated growth of Ag/CuNPs on surface-activated polyacryloamidoxime nanofibers for superior antibacterial activity with improved biocompatibility

Abstract

Introduction: Nanofibers are one of the role-playing innovations of nanotechnology. Their high surface-to-volume ratio allows them to be actively functionalized with a wide range of materials for a variety of applications. The functionalization of nanofibers with different metal nanoparticles (NPs) has been studied widely to fabricate antibacterial substrates to battle antibiotic-resistant bacteria. However, metal NPs show cytotoxicity to living cells, thereby restricting their application in biomedicine.

Objectives: To minimize the cytotoxicity of NPs, biomacromolecule lignin was employed as both a reducing and capping agent to green synthesize silver (Ag) and copper (Cu) NPs on the surface of highly activated polyacryloamidoxime nanofibers. The activation of polyacrylonitrile (PAN) nanofibers via amidoximation was employed for enhanced loading of NPs to achieve superior antibacterial activity.

Methodology: At first, electrospun PAN nanofibers (PANNM) were activated to produce polyacryloamidoxime nanofibers (AO-PANNM) by immersing PANNM in a solution of Hydroxylamine hydrochloride (HH) and Na₂CO₃ under controlled conditions. Later, Ag and Cu ions were loaded by immersing AO-PANNM in different molar concentrations of AgNO₃ and CuSO₄ solutions in a stepwise manner. The reduction of Ag and Cu ions into NPs to fabricate bimetal-coated PANNM (BM-PANNM) was carried out via alkali lignin at 37 °C for 3 h in a shaking incubator with ultrasonication every 1 h.

Results: AO-APNNM and BM-PANNM hold their nano-morphology except for some changes in fiber orientation. XRD analysis demonstrated the formation of Ag and CuNPs as evident from their respective spectral band. Maximum 8.46 ± 0.14 wt% and 0.98 ± 0.04 wt% Ag and Cu species were loaded on AO-PANNM, respectively as revealed by ICP spectrometric analysis. The hydrophobic PANNM turned into super hydrophilic, having WCA of 14 ± 3.32° after amidoximation which further reduced to 0° for BM-PANNM. However, the swelling ratio of PANNM reduced from 13.19 ± 0.18 g/g to 3.72 ± 0.20 g/g for AO-PANNM. Even at the third cycle test against S. aureus strains, 0.1Ag/Cu-PANNM, 0.3Ag/Cu-PANNM, and 0.5Ag/Cu-PANNM displayed bacterial reduction of 71.3 ± 1.64 %, 75.2 ± 1.91 %, and 77.24 ± 1.25 %, respectively. On 3rd cycle test against E. coli, above 82 % bacterial reduction was noticed for all BM-PANNM. Amidoximation increased COS-7 cell viability up to 82 %. The cell viability of 0.1Ag/Cu-PANNM, 0.3Ag/Cu-PANNM, and 0.5Ag/Cu-PANNM was found to be ∼68 %, ∼62, and 54 %, respectively. In LDH assay, almost no release of LDH was detected, suggesting the compatibility of the cell membrane in contact with BM-PANNM. The improved biocompatibility of BM-PANNM even at higher loading (%) of NPs must be ascribed to the controlled release of metal species in the early stage, antioxidant, and biocompatible lignin capping of NPs.

Conclusions: BM-PANNM displayed superior antibacterial activity against E. coli and S. aureus bacterial strains and acceptable biocompatibility of COS-7 cells even at higher loading (%) of Ag/CuNPs. Our findings suggest that BM-PANNM can be used as a potential antibacterial wound dressing and other antibacterial applications where sustained antibacterial activity is needed.

Keywords: Antibacterial activity; Biocompatibility; Controlled release; Lignin-capped nanoparticles; Polyacryloamidoxime nanofibers.