Iron-Doped Apatite Nanoparticles Delivered via Electrospun Fiber Mesh for Maximized Bacterial Killing by Bacteriophage
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Iron-Doped Apatite Nanoparticles Delivered via Electrospun Fiber Mesh for Maximized Bacterial Killing by Bacteriophage Jessica M. Andriolo1, Gary F. Wyss2, John P. Murphy3, Marisa L. Pedulla4, M. Katie Hailer5, Jack L. Skinner6 1 Biomedical Engineering, IIP, University of Montana, Missoula, MT 59812 2 CAMP, Montana Tech, Butte, MT 59701 3 Materials Science, Montana Tech, Butte, MT 59701 4 Biological Sciences, Montana Tech, Butte, MT 59701 5 Chemistry and Geochemistry, Montana Tech, Butte, MT 59701 6 Mechanical Engineering, Montana Tech, Butte, MT 59701 ABSTRACT According to the Centers for Disease Control (CDC) and prevention, at least 2 million people in the United States become infected with antibiotic-resistant bacteria, and at least 23,000 people die each year as a direct result of those infections. One alternative to traditional antibiotics is bacteriophage (phage) therapy. Phage therapy utilizes bacteria-specific viruses to infect and kill bacteria cells. The specificity of these viruses is beneficial in that phage used for therapeutic purposes do not harm the human microbiota, nor do phage infect eukaryotic cells. It has been discovered that iron-doped apatite nanoparticles (IDANPs) significantly enhance phage killing of bacteria cells. The biocompatibility of apatite, coupled with its effectiveness as an adjuvant to enhance an alternative antibiotic therapy, makes it of interest for medical applications. Previously, researchers have encased phage in a microfluidic channel in coaxially electrospun fibers, allowing phage to remain viable after several weeks storage at 4 °C. Here, we have constructed a polymer fiber layer using electrospinning (ES) for delivery of IDANP adjuvants to compliment phage treatment delivery fibers. The IDANP delivery layer constructed is composed of polyethylene oxide (PEO) doped with the nanoparticles. When compared to media-only and IDANP-only controls, results show IDANPs delivered through a PEO fiber mesh remain effective at enhancement of phage infectivity. INTRODUCTION It is well known that bacterial strains have become increasingly resistant to traditional antibiotic therapies, prompting critical research and development of alternatives [1]. As an alternative to traditional antibiotics, researchers have studied the use of bacteria-specific viruses to treat bacterial infection. These bacterial viruses or bacteriophage (phage) adhere to the bacterial surface and inject phage genetic material into the bacterium, thereby manipulating bacterial metabolism, and causing the bacteria to produce phage components within itself [2]. As phage components assemble into complete viral particles, an increase in phage lysin protein accumulates in the bacterial cytosol, eventually hydrolysing peptidoglycan in the bacterial cell wall, thus lysing the bacterial cell, and resulting in a release of mature phage progeny [3, 4]. Newly synthesized phage subsequently kill more bacteria. Phage therapy was studied initially approximately 85 years ago in humans [5], and has demonstrated minimal side effec
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