Fabrication of heteroatom doped NFP-MWCNT and NFB-MWCNT nanocomposite from imidazolium ionic liquid functionalized MWCNT for antibiofilm and wound healing in Wistar rats: Synthesis, characterization, in-vitro and in-vivo studies

Bacterial biofilm is an obstacle for wound healing because it can affect the epithelialization, development of granular cells, and other regular inflammatory procedures. It plays the role of safeguarding pathogens from antiseptics and antibiotics. In this respect, this research work aims to develop heteroatom (N, F, P/B) incorporated mull-walled carbon nanotubes (MWCNT), such as NFP-MWCNT and NFB-MWCNT, which can maximize the wound healing efficacy via destroying the wound pathogen and biofilms. NFP-MWCNT and NFB-MWCNT were obtained using self-assembling ionic liquids (ILs) such as BMIM-PF6 and BMIM-BF4 in an acid-functionalized MWCNT (A-MWCNT) suspension, followed by pyrolysis in a nitrogen atmosphere. The composite formation was established by FTIR, XRD, RAMAN, EDX mapping, and XPS spectroscopy. TEM and SEM analyses confirmed the bamboo stick-like morphology. During this reaction, IL molecules might be cross-linked with A-MWCNT via hydrogen bonding and ionic interaction, with further pyrolysis producing the defects with doping of N, F, P, or B elements. Finally, they were assessed for their antibiofilm activity against typical bacterial strains such as K. pneumoniae, P. aeruginosa, E. coli (Gram-negative), and B. subtilis (Gram-positive), using a quantitative estimation approach. The results revealed greater effectiveness of NFB-MWCNT and NFP-MWCNT, compared to pristine MWCNT. The antibiofilm activity of NFP-MWCNT and NFB-MWCNT was associated with their specific surface chemistry (due to the presence of N, F, P/B heteroatoms), and their nanosize. Moreover, the synthesized material was examined for its wound-healing ability in Wistar rats. The results proved that cells cultured on NFB-MWCNT and NFP-MWCNT displayed exceptional healing ability. The different electronegativity between the heteroatoms creates the surface charge that inhibits the biofilm formation, leading to healing the wounds together with the heteroatom mineral source for mouse fibroblast regeneration and granulation. This is the first study in which the role of different heteroatoms incorporated into MWCNT is examined in the context of antibiofilm-associated wound-healing ability.