mRNA vaccines have the potential to tackle many unmet medical needs that are unable to be addressed with conventional vaccine technologies. A potent and well-tolerated delivery technology is integral to fully realizing the potential of mRNA vaccines. Pre-clinical and clinical studies have demonstrated that mRNA delivered intramuscularly (IM) with first-generation lipid nanoparticles (LNPs) generates robust immune responses. Despite progress made over the past several years, there remains significant opportunity for improvement, as the most advanced LNPs were designed for intravenous (IV) delivery of siRNA to the liver. Here, we screened a panel of proprietary biodegradable ionizable lipids for both expression and immunogenicity in a rodent model when administered IM. A subset of compounds was selected and further evaluated for tolerability, immunogenicity, and expression in rodents and non-human primates (NHPs). A lead formulation was identified that yielded a robust immune response with improved tolerability. More importantly for vaccines, increased innate immune stimulation driven by LNPs does not equate to increased immunogenicity, illustrating that mRNA vaccine tolerability can be improved without affecting potency.

The rising prevalence and severity of antibiotic-resistant biofilm infections poses an alarming threat to public health worldwide. Here, biocompatible multi-compartment nanocarriers were synthesized to contain both hydrophobic superparamagnetic iron oxide nanoparticles (SPIONs) and the hydrophilic antibiotic methicillin for the treatment of medical device-associated infections. SPION co-encapsulation was found to confer unique properties, enhancing both nanocarrier relaxivity and magneticity compared to individual SPIONs. These iron oxide-encapsulating polymersomes (IOPs) penetrated 20 μm thick Staphylococcus epidermidis biofilms with high efficiency following the application of an external magnetic field. Three-dimensional laser scanning confocal microscopy revealed differential bacteria death as a function of drug and SPION loading. Complete eradication of all bacteria throughout the biofilm thickness was achieved using an optimized IOP formulation containing 40 μg/mL SPION and 20 μg/mL of methicillin. Importantly, this formulation was selectively toxic towards methicillin-resistant biofilm cells but not towards mammalian cells. These novel iron oxide-encapsulating polymersomes demonstrate that it is possible to overcome antibiotic-resistant biofilms by controlling the positioning of nanocarriers containing two or more therapeutics.

Here, the antibacterial activity of dextran-coated nanoceria was examined against Pseudomonas aeruginosa and Staphylococcus epidermidis by varying the dose, the time of treatment, and the pH of the solution. Findings suggested that dextran-coated nanoceria particles were much more effective at killing P. aeruginosa and S. epidermidis at basic pH values (pH = 9) compared to acidic pH values (pH = 6) due to a smaller size and positive surface charge at pH 9. At pH 9, different particle concentrations did cause a delay in the growth of P. aeruginosa, whereas impressively S. epidermidis did not grow at all when treated with a 500 μg/mL nanoceria concentration for 24 hours. For both bacteria, a 2 log reduction and elevated amounts of reactive oxygen species (ROS) generation per colony were observed after 6 hours of treatment with nanoceria at pH 9 compared to untreated controls. After 6 hours of incubation with nanoceria at pH 9, P. aeruginosa showed drastic morphological changes as a result of cellular stress. In summary, this study provides significant evidence for the use of nanoceria (+4) for a wide range of anti-infection applications without resorting to the use of antibiotics, for which bacteria are developing a resistance towards anyway.

Hydrogel-based biomaterials have been widely used for tissue engineering applications because of their high water content, swellability, and permeability, which facilitate transport and diffusion of essential nutrients, oxygen, and waste across the scaffold. These characteristics make hydrogels suitable for encapsulating cells and creating a cell supportive environment that promotes tissue regeneration when implanted in vivo. This is particularly important in the context of tissues whose intrinsic regenerative capacity is limited, such as cartilage. However, the clinical translation of hydrogels has been limited by their poor mechanical performance, low adhesive strength, uncontrolled degradation rates, and their susceptibility to bacterial colonization. Here, we introduce an elastic, antimicrobial, and adhesive hydrogel comprised of methacrylated hyaluronic acid (MeHA) and an elastin-like polypeptide (ELP), which can be rapidly photo-cross-linked in situ for the regeneration and repair of different tissues. Hybrid hydrogels with a wide range of physical properties were engineered by varying the concentrations of MeHA and ELP. In addition, standard adhesion tests demonstrated that the MeHA/ELP hydrogels exhibited higher adhesive strength to the tissue than commercially available tissue adhesives. MeHA/ELP hydrogels were then rendered antimicrobial through the incorporation of zinc oxide (ZnO) nanoparticles, and were shown to significantly inhibit the growth of methicillin-resistant Staphylococcus aureus (MRSA), as compared to controls. Furthermore, the composite adhesive hydrogels supported in vitro mammalian cellular growth, spreading, and proliferation. In addition, in vivo subcutaneous implantation demonstrated that MeHA/ELP hydrogels did not elicit any significant inflammatory response, and could be efficiently biodegraded while promoting the integration of new autologous tissue. In summary, we demonstrated for the first time that MeHA/ELP-ZnO hydrogel can be used as an adhesive and antimicrobial biomaterial for tissue engineering applications, because of its highly tunable physical characteristics, as well as remarkable adhesive and antimicrobial properties.

The rapidly diminishing number of effective antibiotics that can be used to treat infectious diseases and associated complications in a physician's arsenal is having a drastic impact on human health today. This study explored the development and optimization of a polymersome nanocarrier formed from a biodegradable diblock copolymer to overcome bacterial antibiotic resistance. Here, polymersomes were synthesized containing silver nanoparticles embedded in the hydrophobic compartment, and ampicillin in the hydrophilic compartment. Results showed for the first time that these silver nanoparticle-embedded polymersomes (AgPs) inhibited the growth of Escherichia coli transformed with a gene for ampicillin resistance (bla) in a dose-dependent fashion. Free ampicillin, AgPs without ampicillin, and ampicillin polymersomes without silver nanoparticles had no effect on bacterial growth. The relationship between the silver nanoparticles and ampicillin was determined to be synergistic and produced complete growth inhibition at a silver-to-ampicillin ratio of 1 : 0.64. In this manner, this study introduces a novel nanomaterial that can effectively treat problematic, antibiotic-resistant infections in an improved capacity which should be further examined for a wide range of medical applications.