Gelatin-coated silicon oxide nanoparticles encapsulated recombinant human secretory leukocyte protease inhibitor (rhSLPI) reduced cardiac cell death against an in vitro simulated ischaemia/reperfusion injury

Abstract

Ischemic Heart Disease (IHD) is the main global cause of death. Previous studies indicated that recombinant human secretory leukocyte protease inhibitor (rhSLPI) exhibits a cardioprotective effect against myocardial ischaemia/reperfusion (I/R) injury. However, SLPI has a short half-life in vivo due to digestion by protease enzymes in circulation. The application of nanoparticle encapsulation could be beneficial for SLPI delivery. Several types of nanoparticles have been developed to encapsulate SLPI and applied in some disease models. However, silica nanoparticles for rhSLPI delivery, particularly on myocardial I/R injury, have never been studied. In this study, we aimed to fabricate gelatin-covered silica nanoparticles (GSNPs) to encapsulate rhSLPI and cardioprotective effect of GSNP-SLPI against an in vitro simulated ischaemia/reperfusion (sI/R). Silica dioxide nanoparticles (SNPs) were fabricated followed by incubation with 0.33 mg/mL of rhSLPI. Then, SNPs containing rhSLPI were coated with gelatin (GSNPs). The GSNPs and rhSLPI-GSNPs were characterized by particle size, zeta potential, and morphology scanning electron microscope (SEM). The concentration of rhSLPI in rhSLPI-GSNPs and drug release was determined by ELISA. Then, cytotoxicity and cardioprotective effect were determined by incubation of GSNPs or rhSLPI-GSNPs with rat cardiac myoblast cell line (H9c2) subjected to simulated ischaemia/reperfusion (sI/R). The results showed the particle size of SNPs, GSNPs, and rhSLPI-GSNPs was 273, 300, and 301 nm, with a zeta potential of −57.21, −22.40, and −24.50 mV, respectively. One milligram of rhSLPI-GSNPs contains 235 ng of rhSLPI. The rhSLPI-GSNPs showed no cytotoxicity on cardiac cells. Treatment with 10 μg/ml of rhSLPI-GSNPs could significantly reduce sI/R induced cardiac cell injury and death. In conclusion, this is the first study to show successful of fabricating novel rhSLPI-encapsulating gelatin-covered silica nanoparticles (rhSLPI-GSNPs) and the cardioprotective effects of rhSLPI-GSNPs against cardiac cell injury and death from myocardial ischaemia/reperfusion.