Abstract: |
Antibiotic resistance poses an important challenge in modern medicine worldwide. A possible strategy to address this issue is the optimization of known antimicrobial compounds, such as curcumin CUR), by improving their bioavailability using nanotechnologies. The aim of this work was to assess the antimicrobial efficacy on Pseudomonas aeruginosa of nanomicelles loaded with CUR alone and activated by blue laser light in an antimicrobial photodynamic therapy (APDT) approach.
The three amphiphilic molecules selected in this study displayed a common C16-long hydrocarbon chain and different amines as the hydrophilic portions: DAPMA (N,N-di-(3-aminopropyl)-N-methylamine), spermidine (SPD) and spermine (SPM). First, free CUR in liquid suspension and loaded in the three amphiphilic nanomicelles were tested with increasing concentrations both on bacteria and keratinocytes to determine both antimicrobial efficacy and safety towards eukaryotic cells. While free CUR exerted limited efficacy showing moderate cytotoxicity, a strong inhibition of bacterial growth was obtained using all three nanosystems without toxicity on eukaryotic cells. CUR-SPM emerged as the most effective, exerting antimicrobial activity up to a 250 nM concentration. Therefore, we selected CUR-SPM for further APDT experiments. We employed three sublethal blue laser (λ 445 nm) protocols (all with 0,1 W/cm2 irradiance, with increasing fluences of 6, 18 and 30 J/cm2) on previously ineffective concentrations of CUR-SPM. We found that the protocols characterized by a fluence of 18 and 30 J/cm2 further decreased the antimicrobial concentration to 50nM. To our knowledge, it is one of the lowest concentrations of CUR with antimicrobial effectiveness among those reported in literature so far.
In the current work, among the CUR-loaded nanomicelles tested, the system based on CUR-loaded SPM nanomicelles showed the strongest antimicrobial effect against P. aeruginosa without cytotoxicity when tested on eukaryotic cells. Moreover, when the CUR-SPM nanomicelles were combined with laser irradiation, the photochemical effectiveness of the natural compound was greatly enhanced. These promising antibacterial results indicate that APDT can be considered as a potential approach to counteract P. aeruginosa infections, although further studies are necessary to unravel the APDT impact at molecular and cellular level.The combination of blue laser APDT with CUR-SPM nanomicelles results in an effective synergistic activity that represents a promising novel therapeutic approach on resistant species. |