Embryonic Stem Cell and Insulin-loaded PLGA Nanoparticles and Evaluation of Efficieny on Pancreatic Beta TC Cells


YÜCEL Ç., YÜCEL Ç., AKTAŞ Y., Değim Z., Yılmaz Ş.

Gazi Pharma Symposium Series, Antalya, Turkey, 12 - 15 November 2015, pp.84

  • Publication Type: Conference Paper / Full Text
  • City: Antalya
  • Country: Turkey
  • Page Numbers: pp.84
  • Erciyes University Affiliated: Yes

Abstract

Diabetes is one of the most common disease that occur globally. In diabetes, insulin is the most effective drug for the control of blood glucose levels [1]. Embryonic stem cells (ESCs) derive from the inner cell mass of the blastocyst, an early-stage embryo. The ESCs subsequently have the potential to differentiate into various types of cells [2]. ESCs have been proposed as a potential source of pancreatic beta cells as well as a powerful tool for cell replacement therapy in the treatment of diabetes [3]. Nanoparticles (NPs) are spherical structures ranging around nanometer in size and prepared from natural and synthetic polymers [4]. In this study, we determinated insulin and ESC-loaded NPs effiencies on diabetic pancreatic beta TC cell line. NP formulations were prepared using poly lactid-co-glycolide acid (PLGA) as a polymer. For amounts of captured insulin and ESC, lowry protein determination method was used. In additon, for ESC encapsulation efficiency, cell counting and western blot were performed. The type of NPs were determined using a transmission electron microscope (TEM). For in vitro characterisation of insulin and ESC NPs, particle size, zeta potential, polydispersity index and encapsulation efficiencies were determined. In vitro insulin release experiment from NPs was performed with a dialysis membrane using Franz-type diffusion cells in pH 7.4 phosphate buffer at 37ºC. In cell culture studies, cytotoxicity test was carried out to determine the toxic effects of insulin and empty nanoparticle. Insulin and empty nanoparticles were also not found to be toxic to cells at any concentrations. Transport experiments of insulin from the NP formulation through pancreatic beta TC cell from the apical side to the basolateral compartment were evaluated. Cumulative amounts of insulin at the end of the 48 h time period were calculated. We investigated the relationship between glucose and insulin concentration in diabetic cell incubated with glucose and streptozocin (STZ). The reduction of insulin concentration was synchronous with increasing glucose level. After applying insulin and ESC loaded NPs to diabetic pancreatic beta TC cells for 48 h, insulin levels were increased for both glucose and STZ-induced diabetic cell groups. It was concluded that insulin and ESC loaded NPs improved the decreased insulin levels inducing glucose and STZ. These NPs may be used in the repair of pancreatic cells and this ESC treatment is a potential source for cell replacement therapy in the treatment of diabetes.