Preparation of arginine-glycine-aspartic acid-modified biopolymeric nanoparticles containing epigalloccatechin-3-gallate for targeting vascular endothelial cells to inhibit corneal neovascularization

Che-Yi Chang; Ming-Chen Wang; Takuya Miyagawa; Zhi-Yu Chen; Feng-Huei Lin; Ko-Hua Chen; Guei-Sheung Liu; Ching-Li Tseng

doi:10.2147/IJN.S114754

Preparation of arginine-glycine-aspartic acid-modified biopolymeric nanoparticles containing epigalloccatechin-3-gallate for targeting vascular endothelial cells to inhibit corneal neovascularization

Int J Nanomedicine. 2016 Dec 30:12:279-294. doi: 10.2147/IJN.S114754. eCollection 2017.

Authors

Che-Yi Chang¹, Ming-Chen Wang², Takuya Miyagawa³, Zhi-Yu Chen³, Feng-Huei Lin⁴, Ko-Hua Chen⁵, Guei-Sheung Liu⁶, Ching-Li Tseng³

Affiliations

¹ Graduate Institute of Biomedical Materials and Tissue Engineering, College of Biomedical Engineering, Taipei Medical University, Taipei; Department of Biomedical Engineering, Chung Yuan Christian University, Taoyuan.
² Department of Biomedical Engineering, Chung Yuan Christian University, Taoyuan.
³ Graduate Institute of Biomedical Materials and Tissue Engineering, College of Biomedical Engineering, Taipei Medical University, Taipei.
⁴ Institute of Biomedical Engineering and Nanomedicine, National Health Research Institutes, Zhunan; Institute of Biomedical Engineering, National Taiwan University.
⁵ Department of Ophthalmology, Taipei Veterans General Hospital; Department of Ophthalmology, School of Medicine, College of Medicine, Taipei Medical University, Taipei, Taiwan.
⁶ Centre for Eye Research Australia, University of Melbourne, Royal Victorian Eye and Ear Hospital, East Melbourne, VIC, Australia.

Abstract

Neovascularization (NV) of the cornea can disrupt visual function, causing ocular diseases, including blindness. Therefore, treatment of corneal NV has a high public health impact. Epigalloccatechin-3-gallate (EGCG), presenting antiangiogenesis effects, was chosen as an inhibitor to treat human vascular endothelial cells for corneal NV treatment. An arginine-glycine-aspartic acid (RGD) peptide-hyaluronic acid (HA)-conjugated complex coating on the gelatin/EGCG self-assembly nanoparticles (GEH-RGD NPs) was synthesized for targeting the α_vβ₃ integrin on human umbilical vein endothelial cells (HUVECs) in this study, and a corneal NV mouse model was used to evaluate the therapeutic effect of this nanomedicine used as eyedrops. HA-RGD conjugation via COOH and amine groups was confirmed by ¹H-nuclear magnetic resonance and Fourier-transform infrared spectroscopy. The average diameter of GEH-RGD NPs was 168.87±22.5 nm with positive charge (19.7±2 mV), with an EGCG-loading efficiency up to 95%. Images of GEH-RGD NPs acquired from transmission electron microscopy showed a spherical shape and shell structure of about 200 nm. A slow-release pattern was observed in the nanoformulation at about 30% after 30 hours. Surface plasmon resonance confirmed that GEH-RGD NPs specifically bound to the integrin α_vβ₃. In vitro cell-viability assay showed that GEH-RGD efficiently inhibited HUVEC proliferation at low EGCG concentrations (20 μg/mL) when compared with EGCG or non-RGD-modified NPs. Furthermore, GEH-RGD NPs significantly inhibited HUVEC migration down to 58%, lasting for 24 hours. In the corneal NV mouse model, fewer and thinner vessels were observed in the alkali-burned cornea after treatment with GEH-RGD NP eyedrops. Overall, this study indicates that GEH-RGD NPs were successfully developed and synthesized as an inhibitor of vascular endothelial cells with specific targeting capacity. Moreover, they can be used in eyedrops to inhibit angiogenesis in corneal NV mice.

Keywords: RGD peptide; antiangiogenesis; corneal neovascularization; epigallocatechin gallate (EGCG); hyaluronic acid (HA); vascular endothelial cells.

MeSH terms

Angiogenesis Inhibitors / administration & dosage
Angiogenesis Inhibitors / pharmacology
Animals
Burns, Chemical / drug therapy
Catechin / administration & dosage
Catechin / analogs & derivatives*
Catechin / chemistry
Cell Survival / drug effects
Corneal Neovascularization / drug therapy*
Corneal Neovascularization / pathology
Disease Models, Animal
Eye Burns / drug therapy
Human Umbilical Vein Endothelial Cells / drug effects
Human Umbilical Vein Endothelial Cells / metabolism
Humans
Hyaluronic Acid / chemistry
Integrin alphaVbeta3 / metabolism
Mice, Inbred C57BL
Molecular Targeted Therapy / methods
Nanoparticles / administration & dosage*
Nanoparticles / chemistry*
Oligopeptides / chemistry*
Ophthalmic Solutions / chemistry
Ophthalmic Solutions / pharmacology

Substances

Angiogenesis Inhibitors
Integrin alphaVbeta3
Oligopeptides
Ophthalmic Solutions
arginyl-glycyl-aspartic acid
Catechin
Hyaluronic Acid
epigallocatechin gallate