Deep Eutectic Solvent Eutectogels for Delivery of Broad-Spectrum Antimicrobials.

Gel-based wound dressings have gained popularity within the healthcare industry for the prevention and treatment of bacterial and fungal infections. Gels based on deep eutectic solvents (DESs), known as eutectogels, provide a promising alternative to hydrogels as they are non-volatile and highly tunable and can solubilize therapeutic agents, including those insoluble in hydrogels. A choline chloride:glycerol-cellulose eutectogel was loaded with numerous antimicrobial agents including silver nanoparticles, black phosphorus nanoflakes, and commercially available pharmaceuticals (octenidine dihydrochloride, tetracycline hydrochloride, and fluconazole). The eutectogels caused >97% growth reduction in Gram-positive methicillin-resistant Staphylococcus aureus and Gram-negative Pseudomonas aeruginosa bacteria and the fungal species Candida albicans.

Deferiprone-Gallium-Protoporphyrin Chitogel Decreases Pseudomonas aeruginosa Biofilm Infection without Impairing Wound Healing.

Pseudomonas aeruginosa is one of the most common pathogens encountered in clinical wound infections. Clinical studies have shown that P. aeruginosa infection results in a larger wound area, inhibiting healing, and a high prevalence of antimicrobial resistance. Hydroxypyridinone-derived iron chelator Deferiprone (Def) and heme analogue Gallium-Protoporphyrin (GaPP) in a chitosan-dextran hydrogel (Chitogel) have previously been demonstrated to be effective against PAO1 and clinical isolates of P. aeruginosa in vitro. Moreover, this combination of these two agents has been shown to improve sinus surgery outcomes by quickly reducing bleeding and preventing adhesions. In this study, the efficacy of Def-GaPP Chitogel was investigated in a P. aeruginosa biofilm-infected wound murine model over 6 days. Two concentrations of Def-GaPP Chitogel were investigated: Def-GaPP high dose (10 mM Def + 500 µg/mL GaPP) and Def-GaPP low dose (5 mM Def + 200 µg/mL GaPP). The high-dose Def-GaPP treatment reduced bacterial burden in vivo from day 2, without delaying wound closure. Additionally, Def-GaPP treatment decreased wound inflammation, as demonstrated by reduced neutrophil infiltration and increased anti-inflammatory M2 macrophage presence within the wound bed to drive wound healing progression. Def-GaPP Chitogel treatment shows promising potential in reducing P. aeruginosa cutaneous infection with positive effects observed in the progression of wound healing.

Novel Antibacterial Materials and Coatings-A Perspective by the Editors.

The fight between humans and bacteria has escalated to a new level.

Therapeutic potential of antimicrobial peptides for treatment of wound infection.

Healing of cutaneous wounds is a fundamental process required to re-establish tissue integrity, repair skin barrier function, and restore skin homeostasis. Chronic wound infection, exacerbated by the growing development of resistance to conventional therapies, hinders the skin repair process and is a serious clinical problem affecting millions of people worldwide. In the past decade, the use of antimicrobial peptides (AMPs) has attracted increasing attention as a potential novel strategy for the treatment of chronic wound infections due to their unique multifaceted mechanisms of action, and AMPs have been demonstrated to function as potent host-defense molecules that can control microbial proliferation, modulate host-immune responses, and act as endogenous mediators of wound healing. To date over 3,200 AMPs have been discovered either from living organisms or through synthetic derivation, some of which have progressed to clinical trials for the treatment of burn and wound injuries. However, progress to routine clinical use has been hindered due to AMPs' susceptibility to wound and environmental factors including changes in pH, proteolysis, hydrolysis, oxidation, and photolysis. This review will discuss the latest research focused on the development and applications of AMPs for wound infections using the latest nanotechnological approaches to improve AMP delivery, and stability to present effective combinatorial treatment for clinical applications.

Bacteria-Activated Dual pH- and Temperature-Responsive Hydrogel for Targeted Elimination of Infection and Improved Wound Healing.

Antibacterial treatment that provides on-demand release of therapeutics that can kill a broad spectrum of pathogens while maintaining long-term efficacy and without developing resistance or causing side effects is urgently required in clinical practice. Here, we demonstrate the development of a multistimuli-responsive hydrogel, prepared by cross-linking N-isopropylacrylamide with acrylic acid and loaded with ultrasmall silver nanoparticles (AgNPs), offering the on-demand release of Ag+ ions triggered by changes in the wound microenvironment. We demonstrate that this dual-responsive hydrogel is highly sensitive to a typical wound pH and temperature change, evidenced by the restricted release of Ag+ ions at acidic pH (<5.5) while significantly promoting the release in alkaline pH (>7.4) (>90% release). The pH-dependent release and antibacterial effect show minimal killing at pH 4 or 5.5 but dramatically activated at pH 7.4 and 10, eliminating >95% of the pathogens. The in vivo antibacterial efficacy and safety showed a high potency to clear Staphylococcus aureus wound infection while significantly accelerating the wound healing rate. This multifunctional hydrogel presents a promising bacteria-responsive delivery platform that serves as an on-demand carrier to not only reduce side effects but also significantly boost the antibacterial efficiency based on physiological needs. It offers great potential to improve the way wound infections are treated with direct clinical implications, providing a single platform for long-lasting application in wound management.

Polycationic Silver Nanoclusters Comprising Nanoreservoirs of Ag+ Ions with High Antimicrobial and Antibiofilm Activity.

Silver-based nano-antibiotics are rapidly developing as promising alternatives to conventional antibiotics. Ideally, to remain potent against a wide range of drug-resistant and anaerobic bacteria, silver-based nano-antibiotics should easily penetrate through the bacterial cell walls and actively release silver ions. In this study, highly monodispersed, ultrasmall (<3 nm), polycationic silver nanoclusters (pAgNCs) are designed and synthesized for the elimination of a range of common Gram-negative and Gram-positive pathogens and their corresponding established and matured biofilms, including those composed of multiple species. The pAgNCs also show greatly enhanced antibacterial efficacy against anaerobic bacteria such as Fusobacterium nucleatum and Streptococcus sanguinis. These results demonstrate that the cationic nature facilitates better penetration to the bacterial cell membrane while the presence of a high percentage (>50%) of silver ions (i.e., Ag+ nanoreservoirs) on the cluster surface maintains their efficiency in both aerobic and anaerobic conditions. Significantly, the pAgNCs showed a strong capacity to significantly delay the development of bacterial resistance when compared to similar-sized negatively charged silver nanoparticles or conventional antibiotics. This study demonstrates a novel design strategy that can lay the foundation for the development of future highly potent nano-antibiotics effective against a broad spectrum of pathogens and biofilms needed in many everyday life applications and industries.

Eradication of Mature Bacterial Biofilms with Concurrent Improvement in Chronic Wound Healing Using Silver Nanoparticle Hydrogel Treatment.

Biofilm-associated infections are a major cause of impaired wound healing. Despite the broad spectrum of anti-bacterial benefits provided by silver nanoparticles (AgNPs), these materials still cause controversy due to cytotoxicity and a lack of efficacy against mature biofilms. Herein, highly potent ultrasmall AgNPs were combined with a biocompatible hydrogel with integrated synergistic functionalities to facilitate elimination of clinically relevant mature biofilms in-vivo combined with improved wound healing capacity. The delivery platform showed a superior release mechanism, reflected by high biocompatibility, hemocompatibility, and extended antibacterial efficacy. In vivo studies using the S. aureus wound biofilm model showed that the AgNP hydrogel (200 µg/g) was highly effective in eliminating biofilm infection and promoting wound repair compared to the controls, including silver sulfadiazine (Ag SD). Treatment of infected wounds with the AgNP hydrogel resulted in faster wound closure (46% closure compared to 20% for Ag SD) and accelerated wound re-epithelization (60% for AgNP), as well as improved early collagen deposition. The AgNP hydrogel did not show any toxicity to tissue and/or organs. These findings suggest that the developed AgNP hydrogel has the potential to be a safe wound treatment capable of eliminating infection and providing a safe yet effective strategy for the treatment of infected wounds.

Multifunctional ultrasmall AgNP hydrogel accelerates healing of S. aureus infected wounds.

The increasing emergence of antibiotic resistance coupled with the limited effectiveness of current treatments highlights the need for the development of new treatment modalities. Silver nanoparticles (AgNPs) are a promising alternative with broad-spectrum antibacterial activity. However, the clinical translation of AgNPs have been hampered primarily due to the delivery of unsafe levels of silver ions (Ag+) resulting in cellular toxicity and their susceptibility to aggregation resulting in loss of efficacy. Here, we describe a safe and effective, thermo-responsive AgNP hydrogel that provides antibacterial effects in conjunction with wound promoting properties. Using a murine model of wound infection, we demonstrate that the applied AgNP hydrogel to the wound (12 µg silver) not only provides superior bactericidal activity but also reduces inflammation leading to accelerated wound closure when compared to industry-standard silver sulfadiazine (302 µg silver). The AgNP hydrogel-treatment significantly accelerated wound closure at day 4 post-infection (56 closure) compared to both blank hydrogel or Ag SD (74% and 91% closure respectively) with a concurrent increase in PCNA-positive proliferating cells corresponding with a significant 32% improvement in wound re-epithelization compared to the blank hydrogel. Treatment of infected wounds with AgNP hydrogel also decreased neutrophil infiltration, increased anti-inflammatory Ym-1 positive M2 macrophages, and reduced the number of caspase-1 positive apoptotic cells. Therefore, this novel multifunctional AgNP thermo-responsive hydrogel is potentially a safe and effective treatment at much lower concentration for the treatment of wound infections. STATEMENT OF SIGNIFICANCE: In this study, we describe the development of a multifunctional thermo-responsive hydrogel of ultrasmall silver nanoparticles (AgNPs) for controlled and optimized delivery of silver to infected wounds. The in vivo biological effects of the developed hydrogel showed significant S. aureus elimination from infected mouse wounds compared to a commercial antibacterial formulation. The developed AgNP hydrogel optimally regulates inflammatory responses to promote wound healing as indicated by increased cell proliferation and wound re-epithelization. Additionally, AgNP hydrogel shows significant potential in regulating neutrophil infiltration while increasing levels of anti-inflammatory M2 macrophages and reduces the number of apoptotic cells. Therefore, the multifunctional properties of the developed AgNP thermo-responsive hydrogel offers great clinical potential to control bacterial infections and promote wound healing.

pH-Responsive "Smart" Hydrogel for Controlled Delivery of Silver Nanoparticles to Infected Wounds.

Persistent wound infections have been a therapeutic challenge for a long time. Current treatment approaches are mostly based on the delivery of antibiotics, but these are not effective for all infections. Here, we report the development of a sensitive pH-responsive hydrogel that can provide controlled, pH-triggered release of silver nanoparticles (AgNPs). This delivery system was designed to sense the environmental pH and trigger the release of AgNPs when the pH changes from acidic to alkaline, as occurs due to the presence of pathogenic bacteria in the wound. Our results show that the prepared hydrogel restricts the release of AgNPs at acidic pH (pH = 4) but substantially amplifies it at alkaline pH (pH = 7.4 and pH = 10). This indicates the potential use of the hydrogel for the on-demand release of Ag+ depending on the environmental pH. In vitro antibacterial studies demonstrated effective elimination of both Gram-negative and positive bacteria. Additionally, the effective antibacterial dose of Ag+ showed no toxicity towards mammalian skin cells. Collectively, this pH-responsive hydrogel presents potential as a promising new material for the treatment of infected wounds.

Quality-by-Design Approach for the Development of Nano-Sized Tea Tree Oil Formulation-Impregnated Biocompatible Gel with Antimicrobial Properties.

Despite the promising properties of tea tree oil (TTO) as potential therapeutics for several superficial skin conditions, certain limitations such as physical instability and skin irritation have restricted its widespread use. This study focuses on developing a rationally designed lipid-based nanoformulation (TTO-LNF) in accordance with the US Food and Drug Administration standard using a well-recognized quality-by-design (QbD) approach. Using a mixture experimental design, TTO-LNF has been optimized with 5% TTO, 10% surfactant, 5% co-surfactant, and 80% water, which showed a 14.4 ± 4.4 nm droplet size and 0.03 ± 0.01 polydispersity index (PDI). To ease the topical administration, the TTO-LNF gel formulation was further developed using xanthan gum to achieve the desired viscosity and form a gel. The in vitro antibacterial tests of TTO-LNF showed promising inhibitory effects toward both Gram-negative and Gram-positive bacteria. In fact, a complete growth inhibition of S. epidermidis was observed when exposed to TTO-LNF and TTO-LNF gel for 24 h, showing better activity than antibiotic kanamycin (25 µg/mL). Additionally, the in vitro release study showed a sustained release profile with a 50% release in 24 h, which could be beneficial to reduce the toxicity and thereby improve the therapeutic efficacy for long-acting applications. Furthermore, the formulations were remarkably stable at 40 °C/75% Relative humidity (RH) for at least 4 weeks. Therefore, this study presents a promising strategy to develop a biocompatible and stable formulation that can be used for the topical treatment of skin infections.

Ultrasmall AgNP-Impregnated Biocompatible Hydrogel with Highly Effective Biofilm Elimination Properties.

Ultrasmall silver nanoparticles (AgNPs; size < 3 nm) have attracted a great deal of interest as an alternative to commercially available antibiotics due to their ability to eliminate a wide range of microbial pathogens. However, most of these ultrasmall AgNPs are highly reactive and unstable, as well as susceptible to fast oxidation. Therefore, both the stability and toxicity remain major shortcomings for their clinical application and uptake. To circumvent these problems, we present a novel strategy to impregnate ultrasmall AgNPs into a biocompatible thermosensitive hydrogel that enables controlled release of silver alongside long-term storage stability and highly potent antibacterial activity. The advantage of this strategy lies in the combination of a homogenous dispersion of AgNPs in a hydrogel network, which serves as a sustained-release reservoir, and the unique feature of ultrasmall AgNP size, which provides an improved biofilm eradication capacity. The superior biofilm dispersion properties of the AgNP hydrogel is demonstrated in both single-species and multispecies biofilms, eradicating ∼80% of established biofilms compared to untreated controls. Notably, the effective antibacterial concentration of the formulation shows minimal toxicity to human fibroblasts and keratinocytes. These findings present a promising novel strategy for the development of AgNP hydrogels as an efficient antibacterial platform to combat resistant bacterial biofilms associated with wound infections.

The interplay between size and valence state on the antibacterial activity of sub-10 nm silver nanoparticles.

Silver nanoparticles (AgNPs) have attracted enormous interest because of their excellent antibacterial properties, low cytotoxicity and limited evidence for resistance. As a general trend, smaller nanoparticles are considered to have stronger antibacterial activity. In this work we investigate whether this trend is valid for the sub-10 nm region by designing and synthesising three types of sub-10 nm AgNPs (∼1.87, ∼2.93 and ∼6.53 nm) to reveal the influence of size, valence state and structure on the antibacterial potency of AgNPs. We found that NPs with a size of ∼2.93 nm having a high concentration of silver in the first valence state presented the highest bacterial killing potency as well as low cytotoxicity to mammalian cells. The new insights presented in this study open future avenues for the engineering of highly potent silver nanoantibiotics that can be incorporated into future advanced medical devices and therapies capable of protecting patients from infections.

Development of Topical Delivery Systems for Flightless Neutralizing Antibody.

Flightless I (Flii) is an actin remodeling protein important for cytoskeletal regulation and cellular processes including migration, proliferation, and adhesion. Previous studies have clearly identified Flii as a novel therapeutical target for improved wound repair and have demonstrated Flii regulation using Flii neutralizing antibodies (FnAb) in different models of wound healing in vivo. Here we describe the development of an optimized topical delivery system that can neutralize Flii activity in the epidermis. Topical delivery of FnAb is an attractive approach as it provides a convenient application, sustained release, localized effect, and reduced dosage. Three successful formulations were developed, and their physical and chemical stability examined. The in vitro release revealed prolonged and sustained release of FnAb in all the tested formulations. Additionally, penetration studies using intact porcine skin showed that FnAb penetrated the epidermis and upper papillary dermis. The penetrated FnAb significantly reduced Flii expression compared to dosed matched IgG controls. This study has successfully developed a topical delivery system for FnAb that could serve as a potential platform for future localized wound treatments.