Development of nanoparticles for delivering recombinant human secretory leukocytes protease inhibitor (rhSLPI) for reducing vascular endothelial cell injury

• Published in: Materials today : proceedings 2023-04
• Main Authors: Phutiyothin, Chayanisa, Chouyratchakarn, Wannapat, Pikwong, Faprathan, Jantrawut, Pensak, Baipaywad, Phornsawat, Kumphune, Sarawut
• Format: Article
• Language: English
• Subjects: Anti-protease; Endothelial cells; Human serum albumin; Ischemia/reperfusion injury; Nanoparticles; Secretory leukocyte protease inhibitor
• ISSN: 2214-7853; 2214-7853
• DOI: 10.1016/j.matpr.2023.04.095

1.4.1.Myocardial ischemia/reperfusion (I/R) is the pathophysiological cause of ischemic heart disease (IHD), which is a leading cause of death globally. I/R injury impacts and damages vascular endothelial cells as well as cardiomyocytes, leading to endothelial dysfunction and then worsened cardiac injury consequently. Previous studies have shown that secretory leukocyte protease inhibitors (SLPI) could provide both cardio- and vasculo-protective benefits against I/R injury. However, SLPI can be destroyed by circulating enzymes and have a short half-life, meaning there are clinical limitations to using SLPI as a therapeutic agent. To overcome these limitations, studies have been conducted on human serum albumin (HSA) nanoparticles for recombinant human SLPI delivery, but not for vascular endothelial cells subjected to I/R injury. In this study, HSA nanoparticles were fabricated using the nanoprecipitation method. Subsequently, rhSLPI-HSA nanoparticles were generated by adding 150 µl SLPI (0.75 µg/µl) to human serum albumin to produce particles. Then, the size and zeta potential of blank-HSA and rhSLPI nanoparticles were measured by field emission scanning electron microscopy (FE-SEM) and dynamic light scattering (DLS). The results showed that the size of blank-HSA nanoparticles and rhSLPI-nanoparticles was 147 nm and 200–300 nm, respectively. The zeta potential of blank-HSA nanoparticles and rhSLPI-nanoparticles was −16.9 mV and −13 mV, respectively. Afterward, the cytotoxicity of both types of nanoparticles was examined on human vascular endothelial cell lines (EA.hy 926) by an MTT assay. The results showed that there was no toxicity in either type of nanoparticle on the vascular endothelial cells. The optimal concentrations for rhSLPI-HSA-NPs were 1 ug/ml and 10 ug/ml, which could significantly increase cell viability. The current study demonstrated the possibility of being able to generate human serum albumin nanoparticles encapsulated with recombinant human SLPI (rhSLPI-HSA nanoparticles) with no cytotoxicity, which could be used for determining the vasculo-protective effect of rhSLPI-HSA nanoparticles against an in vitro simulated I/R-induced vascular endothelial cell injury.