Antibacterial, Anti-Biofilm and Pro-Migratory Effects of Double Layered Hydrogels Packaged with Lactoferrin-DsiRNA-Silver Nanoparticles for Chronic Wound Therapy

Abstract

Antimicrobial resistance and biofilm formation in diabetic foot infections worsened during the COVID-19 pandemic, resulting in more severe infections and increased amputations. Therefore, this study aimed to develop a dressing that could effectively aid in the wound healing process and prevent bacterial infections by exerting both antibacterial and anti-biofilm effects. Silver nanoparticles (AgNPs) and lactoferrin (LTF) have been investigated as alternative antimicrobial and anti-biofilm agents, respectively, while dicer-substrate short interfering RNA (DsiRNA) has also been studied for its wound healing effect in diabetic wounds. In this study, AgNPs were complexed with LTF and DsiRNA via simple complexation before packaging in gelatin hydrogels. The formed hydrogels exhibited 1668% maximum swellability, with a 46.67 ± 10.33 µm average pore size. The hydrogels demonstrated positive antibacterial and anti-biofilm effects toward the selected Gram-positive and Gram-negative bacteria. The hydrogel containing AgLTF at 125 µg/mL was also non-cytotoxic on HaCaT cells for up to 72 h of incubation. The hydrogels containing DsiRNA and LTF demonstrated superior pro-migratory effects compared to the control group. In conclusion, the AgLTF-DsiRNA-loaded hydrogel possessed antibacterial, anti-biofilm, and pro-migratory activities. These findings provide a further understanding and knowledge on forming multipronged AgNPs consisting of DsiRNA and LTF for chronic wound therapy.

Metal-Based Nanoparticles as Antimicrobial Agents: A Review

“…On the other hand, silver-based antimicrobial agents have been prepared with the development of AgNPs and nanocluster preparation techniques. These AgNPs have an ultrasmall hydrodynamic diameter (HD) (smaller than 5.5 nm, the renal filtration threshold) and can be rapidly eliminated through the urine system in a reasonable time, reducing the risk of long-term cytotoxicity. − Long-term studies have demonstrated that metallic silver possesses antimicrobial characteristics, and a recent study has shown that low concentrations of silver are not harmful to human cells. , The main cause of AgNP toxicity is their accumulation in cell culture media, cellular uptake, and intracellular localization . According to the results of the current study, numerous variables affect the dangerous limit concentration of AgNPs, including their size, shape, concentration, and surface area.…”

“…138−142 Long-term studies have demonstrated that metallic silver possesses antimicrobial characteristics, and a recent study has shown that low concentrations of silver are not harmful to human cells. 143,144 The main cause of AgNP toxicity is their accumulation in cell culture media, cellular uptake, and intracellular localization. 145 According to the results of the current study, numerous variables affect the dangerous limit concentration of AgNPs, including their size, shape, concentration, and surface area.…”

Metal Nanoparticles: Advanced and Promising Technology in Diabetic Wound Therapy

“…However, Fathil et al successfully formed AgLTF by complexing AgNPs with LTF and utilized this for the targeted therapy of diabetic wounds by loading DsiRNA. 104 The negatively charged DsiRNA was adsorbed onto AgLTF to silence the prostaglandin transporter (PGT) and promote wound healing. Although this study did not employ AgNPs as a drug carrier, the successful adsorption of DsiRNA onto AgLTF demonstrates the potential use of AgNPs as carriers in future applications beyond their antibacterial properties.…”

Hydrogel-based dressings designed to facilitate wound healing

“…To address this issue, researchers have successfully developed various novel and effective antimicrobial hydrogels. 281–283 Cai et al developed a eucalyptus essential oil nanoemulsion hydrogel specifically for this purpose. In vitro experiments demonstrated that the hydrogel displayed strong antimicrobial activity and inhibited biofilm formation.…”