title: Preparation of arginine-glycine-aspartic acid-modified biopolymeric nanoparticles containing epigalloccatechin-3-gallate for targeting vascular endothelial cells to inhibit corneal neovascularization.
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 αvβ3integrin 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 by1H-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 αvβ3. 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.
authors.affiliation: 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.
authors.forename: Che-Yi
authors.initials: CY
authors.lastname: Chang
authors.affiliation: Department of Biomedical Engineering, Chung Yuan Christian University, Taoyuan.
authors.forename: Ming-Chen
authors.initials: MC
authors.lastname: Wang
authors.affiliation: Graduate Institute of Biomedical Materials and Tissue Engineering, College of Biomedical Engineering, Taipei Medical University, Taipei.
authors.forename: Takuya
authors.initials: T
authors.lastname: Miyagawa
authors.affiliation: Graduate Institute of Biomedical Materials and Tissue Engineering, College of Biomedical Engineering, Taipei Medical University, Taipei.
authors.forename: Zhi-Yu
authors.initials: ZY
authors.lastname: Chen
authors.affiliation: Institute of Biomedical Engineering and Nanomedicine, National Health Research Institutes, Zhunan; Institute of Biomedical Engineering, National Taiwan University.
authors.forename: Feng-Huei
authors.initials: FH
authors.lastname: Lin
authors.affiliation: Department of Ophthalmology, Taipei Veterans General Hospital; Department of Ophthalmology, School of Medicine, College of Medicine, Taipei Medical University, Taipei, Taiwan.
authors.forename: Ko-Hua
authors.initials: KH
authors.lastname: Chen
authors.affiliation: Centre for Eye Research Australia, University of Melbourne, Royal Victorian Eye and Ear Hospital, East Melbourne, VIC, Australia.
authors.forename: Guei-Sheung
authors.initials: GS
authors.lastname: Liu
authors.affiliation: Graduate Institute of Biomedical Materials and Tissue Engineering, College of Biomedical Engineering, Taipei Medical University, Taipei.
authors.forename: Ching-Li
authors.initials: CL
authors.lastname: Tseng
mesh_terms: D020533:Angiogenesis Inhibitors / Q000008:administration & dosage / Q000494:pharmacology; D000818:Animals; D002057:Burns, Chemical / Q000188:drug therapy; D002392:Catechin / Q000008:administration & dosage / Q000031:analogs & derivatives* / Q000737:chemistry; D002470:Cell Survival / Q000187:drug effects; D016510:Corneal Neovascularization / Q000188:drug therapy* / Q000473:pathology; D004195:Disease Models, Animal; D005126:Eye Burns / Q000188:drug therapy; D061307:Human Umbilical Vein Endothelial Cells / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D006820:Hyaluronic Acid / Q000737:chemistry; D039302:Integrin alphaVbeta3 / Q000378:metabolism; D008810:Mice, Inbred C57BL; D058990:Molecular Targeted Therapy / Q000379:methods; D053758:Nanoparticles / Q000008:administration & dosage* / Q000737:chemistry*; D009842:Oligopeptides / Q000737:chemistry*; D009883:Ophthalmic Solutions / Q000737:chemistry / Q000494:pharmacology
chemical_list: D020533:Angiogenesis Inhibitors; D039302:Integrin alphaVbeta3; D009842:Oligopeptides; D009883:Ophthalmic Solutions; C047981:arginyl-glycyl-aspartic acid; D002392:Catechin; D006820:Hyaluronic Acid; C045651:epigallocatechin gallate
keywords: RGD peptide; antiangiogenesis; corneal neovascularization; epigallocatechin gallate (EGCG); hyaluronic acid (HA); vascular endothelial cells