Remarkable Effect of Jacalin in Diminishing the Protein Corona Interference in the Antibacterial Activity of Pectin-Capped Copper Sulfide Nanoparticles

Abstract

Herein, we report a new strategy based on jacalin functionalization to diminish the impact of biological fluids in the antibacterial applications of nanoparticles (NPs). Precoating pectin-capped copper sulfide NPs (pCuS) with bovine serum albumin produced a protein corona, which affects the antibacterial activity of pCuS. It was found that the minimum inhibitory concentration (MIC) increases fourfold because of the formation of the protein corona. Interestingly, the pCuS functionalized with jacalin enhance the targeting capabilities through bacterial cell surface glycan recognition with no interference from the protein corona. The MIC of pCuS decreases 16-fold on functionalization with jacalin. Mechanistic studies indicated that the pCuS functionalized with jacalin impede the protein corona interference and induce bacterial cell death by impairing the GSH/reactive oxygen species balance and disrupting the bacteria cell membrane. As a proof of concept, we used a bacteria-infected zebrafish animal model to demonstrate the interference of biological fluids in the antibacterial activity of NPs. Infected zebrafish treated with 1× MIC of pCuS failed to recover from the infection, but 4× MIC rescues the fish. The requirement of a high dose of NPs to treat the infection confirms the interference of biological fluids in nanotherapeutic applications. At the same time, the jacalin–pCuS complex rescues the infected fish at 16-fold lesser MIC. The results obtained from this study suggest that jacalin-mediated NP targeting may have broad implications in the development of future nanomedicine.

Shrimp lectin (Md-Lec) conjugated copper sulfide nanoparticles enhance the elimination of aquatic pathogens in infected Nile tilapia (Oreochromis niloticus)

“…In REMA, the respiring viable cells reduce resazurin to resorun and appear pink in color, whereas the cells lacking respiration appear blue in color. 23 It is noted from Fig. 3, the MIC of pCuS NPs is 12.5 mM against the tested pathogens as they appear blue in colour, suggesting that the cells are inactive; below that, the cells are able to respire and convert the resazurin to resorun.…”

“…The CuS NPs were prepared using the procedure reported in Subramaniyan et al 23 Briey, pectin (50 mg) and copper chloride (1 mM) were dissolved in 50 ml double distilled water. To this mixture, 200 ml ammonium hydroxide was added, which was followed by 400 ml hydrazine hydride.…”

“…The minimum inhibitory concentration of pCuS NPs and the complex, Md-Lec-pCuS NPs against three aquatic pathogens, Aeromonas hydrophila (ATCC 7966), Enterococcus fecalis (ATCC 29212) and Streptococcus iniae (ATCC 29178) were determined by REMA method. 23,25 Briey, 100 ml pCuS NPs (100 mM) was added to rst well of the 96-well plate and then serially diluted. Next, each well was mixed with Md-Lec (25 mg ml À1 ).…”

“…40 Infection was done intramuscularly and treatment was given from the opposite side to that of the infection. 23 The in vivo bacterial infection and the treatment with Md-Lec-pCuS NPs (1Â MIC) were performed in Nile tilapia that was divided into 3 groups; group I: bacteria infected control, group II: bacterial infected + treated with Md-Lec-pCuS NPs, group III: control, uninfected sh. Each group consisted of 10 sh.…”

“…These results go in parallel with the LPO activity reported earlier. 23,33 3.8 In vivo antibacterial studies…”