Thymol polymeric nanoparticle synthesis and its effects on the toxicity of high glucose on OEC cells: involvement of growth factors and integrin‐linked kinase

Abstract

Background

Nowadays, the drug delivery system is important in the treatment of diseases.

Purpose

A polymeric nanoparticle modified by oleic acid (NPMO) as a Thymol (Thy) drug release system was synthesized from Thymbra spicata and its neurotrophic and angiogenic effects on rat’s olfactory ensheathing cells (OECs) in normal (NG) and high glucose (HG) conditions were studied.

Methods

The NPMO was characterized by using different spectroscopy methods, such as infrared, HNMR, CNMR, gel permeation chromatography, dynamic light scattering, and atomic force microscopy. Load and releasing were investigated by HPLC. The toxicity against OECs diet-induced by MTT assay. ROS and generation of nitric oxide (NO) were evaluated using dichloro-dihydro-fluorescein and Griess method, respectively. The expression of protein integrin-linked kinase (ILK), vascular endothelial growth factor (VEGF), brain-derived neurotrophic factor (BDNF), and nerve growth factor (NGF) were evaluated by Western blotting.

Results

ThyNPMO is desirable for transferring drug as a carrier. The amount of Thy and extract (E) loaded on NPMO estimated at 43±2.5% and 41±1.8%, respectively. Then, 65% and 63% of the drug load were released, respectively. Thy, ThyNPMO, E, and ENPMO prevented HG-induced OECs cell death (EC50 33±1.5, 22±0.9, 35±1.8, and 25±1.1 μM, respectively). Incubation with Thy, ThyNPMO, E ,and ENPMO at high concentrations increased cell death with LC50 105±3.5, 82±2.8, 109±4.3, and 86±3.4 μM, respectively in HG states.

Conclusion

OECs were protected by ThyNPMO and ENPMO in protective concentrations by reducing the amount of ROS and NO, maintaining ILK, reducing VEGF, and increasing BDNF and NGF. The mentioned mechanisms were totally reversed at high concentrations.

Keywords:

• thymol
• polymeric nanoparticle
• olfactory enshealing cell
• drug delivery
• high glucose

Acknowledgments

This study was supported by Islamic Azad University, Ilam branch, Iran and Ilam University of Medical Sciences (grant no. 87-04-33-8073).

Abbreviation list

HG, high glucose; NG, normal glucose; HM, high mannitol; Thy, Thymol; NPMO, polymeric nanoparticles modified by oleic acid; E, extract; OEC, olfactory ensheathing cells; G, glycerol; CA, citric acid; NMR, nuclear magnetic resonance; FTIR, Fourier-transform infrared; GPC, gel permeation chromatography; AFM, atomic force microscopy; DLS, dynamic light scattering; NO, nitric oxide; ILK, integrin-linked kinase; VEGF, vascular endothelial growth factor; BDNF, brain-derived neurotrophic factor; NGF, nerve growth factor.

Disclosure

The authors report no conflicts of interest in this work.