Investigation for the pro-angiogenic properties of eggshell-derived nanoparticles

Eggshells are regular domestic, agricultural waste, and the primary composition of eggshells is majorly of calcium, as well as trace amounts of magnesium and phosphorous. These two elements (calcium and magnesium) are also present in living organisms and play an important role in many biological processes (cell growth, muscle contraction, glycolysis, angiogenesis, and vasculogenesis). Considering their role in different biological processes, especially in angiogenesis (formation of new blood vessels from pre-existing vasculature), we hypothesized the involvement of calcium and magnesium (present in eggshells) in the nanoform may induce angiogenesis. To this context, the present manuscript attempts to design calcium-rich nanoparticles derived from both unfertilized and fertilized eggshells (U-ES and F-ES), and investigate their pro-angiogenic properties. Both U-ES nanoparticles (U-ES-NP) and F-ES nanoparticles (F-ES-NP) were developed by the calcination of raw eggshells. These nanoparticles (U-ES-NP and F-ES-NP) are characterized using various analytical techniques. These nanoparticles exhibit pro-angiogenic properties, as validated by<i>in vitro</i>assays (cell proliferation assay, tube formation assay, etc),<i>ex vivo</i>(chick aorta assay) and<i>in vivo</i>(chick choriallantoic membrane assay) experiments. The hemolysis experiment (<i>ex vivo</i>) was performed by incubating mouse RBCs with nanoparticles, which further validates the biocompatibility of these nanomaterials. Taking these results altogether, the current study demonstrates pro-angiogenic properties of biocompatible ES-NP, that could be further utilized for the treatment of several diseases and other biomedical applications after proper biosafety evaluation.