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Therapeutic effect of apatinib-loaded nanoparticles on diabetes-induced retinal vascular leakage.

Jeong JH, Nguyen HK, Lee JE, Suh W - Int J Nanomedicine (2016)

Bottom Line: In addition, they substantially reduced the VEGF-induced junctional loss and internalization of vascular endothelial-cadherin, a major component of endothelial junction complexes.These in vitro and in vivo data indicated that Apa-HSA-PEG nanoparticles efficiently blocked VEGF-induced breakdown of the blood-retinal barrier.These results demonstrated, for the first time, that apatinib-loaded nanoparticles may be a promising therapeutic agent for the prevention and treatment of diabetes-induced retinal vascular disorders.

View Article: PubMed Central - PubMed

Affiliation: School of Pharmacy, Sungkyunkwan University, Suwon.

ABSTRACT
Apatinib, a novel and selective inhibitor of vascular endothelial growth factor (VEGF) receptor 2, has been demonstrated recently to exhibit anticancer efficacy by inhibiting the VEGF signaling pathway. Given the importance of VEGF in retinal vascular leakage, the present study was designed to investigate whether apatinib-loaded polymeric nanoparticles inhibit VEGF-mediated retinal vascular hyperpermeability and block diabetes-induced retinal vascular leakage. For the delivery of water-insoluble apatinib, the drug was encapsulated in nanoparticles composed of human serum albumin (HSA)-conjugated polyethylene glycol (PEG). In vitro paracellular permeability and transendothelial electric resistance assays showed that apatinib-loaded HSA-PEG (Apa-HSA-PEG) nanoparticles significantly inhibited VEGF-induced endothelial hyperpermeability in human retinal microvascular endothelial cells. In addition, they substantially reduced the VEGF-induced junctional loss and internalization of vascular endothelial-cadherin, a major component of endothelial junction complexes. In vivo intravitreal injection of Apa-HSA-PEG nanoparticles in mice blocked VEGF-induced retinal vascular leakage. These in vitro and in vivo data indicated that Apa-HSA-PEG nanoparticles efficiently blocked VEGF-induced breakdown of the blood-retinal barrier. In vivo experiments with streptozotocin-induced diabetic mice showed that an intravitreal injection of Apa-HSA-PEG nanoparticles substantially inhibited diabetes-induced retinal vascular leakage. These results demonstrated, for the first time, that apatinib-loaded nanoparticles may be a promising therapeutic agent for the prevention and treatment of diabetes-induced retinal vascular disorders.

No MeSH data available.


Related in: MedlinePlus

Apa-HSA-PEG nanoparticles block VEGF-induced hyperpermeability in HRMECs.Notes: (A) Cytotoxicity of Apa-HSA-PEG nanoparticles and apatinib was evaluated using the Cell Counting Kit-8 assay (n=6). HRMECs were incubated with apatinib (1 and 5 µM) or Apa-HSA-PEG nanoparticles (960 and 4.8 µg corresponding to 1 and 5 µM apatinib, respectively) for 1–2 days. (B and C) Endothelial permeability was evaluated by measuring the (B) passage of FITC-dextran and (C) TEER in a HRMEC monolayer. HRMECs pretreated with apatinib (1 µM) or Apa-HSA-PEG nanoparticles (6.1 µg corresponding to 1 µM apatinib), and untreated cells were stimulated with rhVEGF (50 ng/mL). (B) FITC-dextran permeability is expressed as the fold change ± SEM with respect to the PBS control (*P<0.05 vs PBS control, #P<0.05 vs VEGF, n=6). (C) In the TEER experiment, specified reagents were added to the upper chamber at time zero, and serial changes in electrical resistance were measured 30 and 60 minutes later. Electrical resistance is expressed as the fold change ± SEM relative to the PBS control at each time point (*P<0.05 vs PBS control, #P<0.05 vs VEGF, n=6).Abbreviations: Apa-HSA-PEG, apatinib-loaded human serum albumin-conjugated polyethylene glycol; VEGF, vascular endothelial growth factor; HRMECs, human retinal microvascular endothelial cells; FITC, fluorescein isothiocyanate; TEER, transendothelial electrical resistance; rhVEGF, recombinant human VEGF; SEM, standard error of the mean; PBS, phosphate-buffered saline; NS, nonsignificant.
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f2-ijn-11-3101: Apa-HSA-PEG nanoparticles block VEGF-induced hyperpermeability in HRMECs.Notes: (A) Cytotoxicity of Apa-HSA-PEG nanoparticles and apatinib was evaluated using the Cell Counting Kit-8 assay (n=6). HRMECs were incubated with apatinib (1 and 5 µM) or Apa-HSA-PEG nanoparticles (960 and 4.8 µg corresponding to 1 and 5 µM apatinib, respectively) for 1–2 days. (B and C) Endothelial permeability was evaluated by measuring the (B) passage of FITC-dextran and (C) TEER in a HRMEC monolayer. HRMECs pretreated with apatinib (1 µM) or Apa-HSA-PEG nanoparticles (6.1 µg corresponding to 1 µM apatinib), and untreated cells were stimulated with rhVEGF (50 ng/mL). (B) FITC-dextran permeability is expressed as the fold change ± SEM with respect to the PBS control (*P<0.05 vs PBS control, #P<0.05 vs VEGF, n=6). (C) In the TEER experiment, specified reagents were added to the upper chamber at time zero, and serial changes in electrical resistance were measured 30 and 60 minutes later. Electrical resistance is expressed as the fold change ± SEM relative to the PBS control at each time point (*P<0.05 vs PBS control, #P<0.05 vs VEGF, n=6).Abbreviations: Apa-HSA-PEG, apatinib-loaded human serum albumin-conjugated polyethylene glycol; VEGF, vascular endothelial growth factor; HRMECs, human retinal microvascular endothelial cells; FITC, fluorescein isothiocyanate; TEER, transendothelial electrical resistance; rhVEGF, recombinant human VEGF; SEM, standard error of the mean; PBS, phosphate-buffered saline; NS, nonsignificant.

Mentions: We first determined the cytotoxicity of Apa-HSA-PEG nanoparticles in HRMECs. Apa-HSA-PEG nanoparticles did not compromise the viability of HRMECs (Figure 2A). We then evaluated the inhibitory effect of Apa-HSA-PEG nanoparticles on VEGF-induced hyperpermeability in HRMECs. VEGF significantly enhanced the passage of FITC-labeled dextran across the HRMEC monolayer (Figure 2B). At a noncytotoxic concentration (1 µM on an apatinib basis), Apa-HSA-PEG nanoparticles completely inhibited VEGF-induced endothelial hyperpermeability. The inhibitory effect of Apa-HSA-PEG nanoparticles was similar to that of free apatinib solution at the same concentration. The TEER assay also demonstrated the ability of Apa-HSA-PEG nanoparticles to block the VEGF-mediated decrease in electrical resistance across the HRMEC monolayer (Figure 2C).


Therapeutic effect of apatinib-loaded nanoparticles on diabetes-induced retinal vascular leakage.

Jeong JH, Nguyen HK, Lee JE, Suh W - Int J Nanomedicine (2016)

Apa-HSA-PEG nanoparticles block VEGF-induced hyperpermeability in HRMECs.Notes: (A) Cytotoxicity of Apa-HSA-PEG nanoparticles and apatinib was evaluated using the Cell Counting Kit-8 assay (n=6). HRMECs were incubated with apatinib (1 and 5 µM) or Apa-HSA-PEG nanoparticles (960 and 4.8 µg corresponding to 1 and 5 µM apatinib, respectively) for 1–2 days. (B and C) Endothelial permeability was evaluated by measuring the (B) passage of FITC-dextran and (C) TEER in a HRMEC monolayer. HRMECs pretreated with apatinib (1 µM) or Apa-HSA-PEG nanoparticles (6.1 µg corresponding to 1 µM apatinib), and untreated cells were stimulated with rhVEGF (50 ng/mL). (B) FITC-dextran permeability is expressed as the fold change ± SEM with respect to the PBS control (*P<0.05 vs PBS control, #P<0.05 vs VEGF, n=6). (C) In the TEER experiment, specified reagents were added to the upper chamber at time zero, and serial changes in electrical resistance were measured 30 and 60 minutes later. Electrical resistance is expressed as the fold change ± SEM relative to the PBS control at each time point (*P<0.05 vs PBS control, #P<0.05 vs VEGF, n=6).Abbreviations: Apa-HSA-PEG, apatinib-loaded human serum albumin-conjugated polyethylene glycol; VEGF, vascular endothelial growth factor; HRMECs, human retinal microvascular endothelial cells; FITC, fluorescein isothiocyanate; TEER, transendothelial electrical resistance; rhVEGF, recombinant human VEGF; SEM, standard error of the mean; PBS, phosphate-buffered saline; NS, nonsignificant.
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f2-ijn-11-3101: Apa-HSA-PEG nanoparticles block VEGF-induced hyperpermeability in HRMECs.Notes: (A) Cytotoxicity of Apa-HSA-PEG nanoparticles and apatinib was evaluated using the Cell Counting Kit-8 assay (n=6). HRMECs were incubated with apatinib (1 and 5 µM) or Apa-HSA-PEG nanoparticles (960 and 4.8 µg corresponding to 1 and 5 µM apatinib, respectively) for 1–2 days. (B and C) Endothelial permeability was evaluated by measuring the (B) passage of FITC-dextran and (C) TEER in a HRMEC monolayer. HRMECs pretreated with apatinib (1 µM) or Apa-HSA-PEG nanoparticles (6.1 µg corresponding to 1 µM apatinib), and untreated cells were stimulated with rhVEGF (50 ng/mL). (B) FITC-dextran permeability is expressed as the fold change ± SEM with respect to the PBS control (*P<0.05 vs PBS control, #P<0.05 vs VEGF, n=6). (C) In the TEER experiment, specified reagents were added to the upper chamber at time zero, and serial changes in electrical resistance were measured 30 and 60 minutes later. Electrical resistance is expressed as the fold change ± SEM relative to the PBS control at each time point (*P<0.05 vs PBS control, #P<0.05 vs VEGF, n=6).Abbreviations: Apa-HSA-PEG, apatinib-loaded human serum albumin-conjugated polyethylene glycol; VEGF, vascular endothelial growth factor; HRMECs, human retinal microvascular endothelial cells; FITC, fluorescein isothiocyanate; TEER, transendothelial electrical resistance; rhVEGF, recombinant human VEGF; SEM, standard error of the mean; PBS, phosphate-buffered saline; NS, nonsignificant.
Mentions: We first determined the cytotoxicity of Apa-HSA-PEG nanoparticles in HRMECs. Apa-HSA-PEG nanoparticles did not compromise the viability of HRMECs (Figure 2A). We then evaluated the inhibitory effect of Apa-HSA-PEG nanoparticles on VEGF-induced hyperpermeability in HRMECs. VEGF significantly enhanced the passage of FITC-labeled dextran across the HRMEC monolayer (Figure 2B). At a noncytotoxic concentration (1 µM on an apatinib basis), Apa-HSA-PEG nanoparticles completely inhibited VEGF-induced endothelial hyperpermeability. The inhibitory effect of Apa-HSA-PEG nanoparticles was similar to that of free apatinib solution at the same concentration. The TEER assay also demonstrated the ability of Apa-HSA-PEG nanoparticles to block the VEGF-mediated decrease in electrical resistance across the HRMEC monolayer (Figure 2C).

Bottom Line: In addition, they substantially reduced the VEGF-induced junctional loss and internalization of vascular endothelial-cadherin, a major component of endothelial junction complexes.These in vitro and in vivo data indicated that Apa-HSA-PEG nanoparticles efficiently blocked VEGF-induced breakdown of the blood-retinal barrier.These results demonstrated, for the first time, that apatinib-loaded nanoparticles may be a promising therapeutic agent for the prevention and treatment of diabetes-induced retinal vascular disorders.

View Article: PubMed Central - PubMed

Affiliation: School of Pharmacy, Sungkyunkwan University, Suwon.

ABSTRACT
Apatinib, a novel and selective inhibitor of vascular endothelial growth factor (VEGF) receptor 2, has been demonstrated recently to exhibit anticancer efficacy by inhibiting the VEGF signaling pathway. Given the importance of VEGF in retinal vascular leakage, the present study was designed to investigate whether apatinib-loaded polymeric nanoparticles inhibit VEGF-mediated retinal vascular hyperpermeability and block diabetes-induced retinal vascular leakage. For the delivery of water-insoluble apatinib, the drug was encapsulated in nanoparticles composed of human serum albumin (HSA)-conjugated polyethylene glycol (PEG). In vitro paracellular permeability and transendothelial electric resistance assays showed that apatinib-loaded HSA-PEG (Apa-HSA-PEG) nanoparticles significantly inhibited VEGF-induced endothelial hyperpermeability in human retinal microvascular endothelial cells. In addition, they substantially reduced the VEGF-induced junctional loss and internalization of vascular endothelial-cadherin, a major component of endothelial junction complexes. In vivo intravitreal injection of Apa-HSA-PEG nanoparticles in mice blocked VEGF-induced retinal vascular leakage. These in vitro and in vivo data indicated that Apa-HSA-PEG nanoparticles efficiently blocked VEGF-induced breakdown of the blood-retinal barrier. In vivo experiments with streptozotocin-induced diabetic mice showed that an intravitreal injection of Apa-HSA-PEG nanoparticles substantially inhibited diabetes-induced retinal vascular leakage. These results demonstrated, for the first time, that apatinib-loaded nanoparticles may be a promising therapeutic agent for the prevention and treatment of diabetes-induced retinal vascular disorders.

No MeSH data available.


Related in: MedlinePlus