RESUMO
PURPOSE: To determine whether intravitreal vasohibin-1 will reduce the grade of the choroidal neovascularization in monkey eyes. METHODS: Choroidal neovascularizations were induced in 12 monkey eyes by laser photocoagulation. Three monkeys were evaluated for the safety of the vasohibin-1 injections, 6 monkeys for the effects of a single injection, and 3 monkeys for repeated injections of vasohibin-1. Ophthalmoscopy, fluorescein angiography, focal electroretinograms, and optical coherence tomography were used for the evaluations. The level of vascular endothelial growth factor in the aqueous was determined by enzyme-linked immunosorbent assay. Immunohistochemistry was performed. RESULTS: An intravitreal injection of 10 µg of vasohibin-1 induced mild intraocular inflammation. Eyes with an intravitreal injection of 0.1 µg and 1.0 µg of vasohibin-1 had significant less fluorescein leakage from the choroidal neovascularizations and larger amplitude focal electroretinograms than that of vehicle-injected eyes. Similar results were obtained by repeated injections of 0.1 µg of vasohibin-1. Immunohistochemistry showed that vasohibin-1 was expressed mainly in the endothelial cells within the choroidal neovascularizations. The vascular endothelial growth factor level was not significantly altered by intravitreal vasohibin-1. CONCLUSION: The reduction of the laser-induced choroidal neovascularizations and preservation of macular function in monkey by intravitreal vasohibin-1 suggest that it should be considered for suppressing choroidal neovascularizations in humans.
Assuntos
Inibidores da Angiogênese/administração & dosagem , Proteínas de Ciclo Celular/administração & dosagem , Neovascularização de Coroide/tratamento farmacológico , Inibidores da Angiogênese/metabolismo , Animais , Proteínas de Ciclo Celular/metabolismo , Neovascularização de Coroide/metabolismo , Neovascularização de Coroide/patologia , Neovascularização de Coroide/fisiopatologia , Modelos Animais de Doenças , Eletrorretinografia/efeitos dos fármacos , Ensaio de Imunoadsorção Enzimática , Angiofluoresceinografia , Imuno-Histoquímica , Injeções Intravítreas , Macaca , Oftalmoscopia , Tomografia de Coerência Óptica , Fator A de Crescimento do Endotélio Vascular/metabolismoRESUMO
In this study, we prepared injectable collagen microspheres for the sustained delivery of recombinant human vascular endothelial growth factor (rhVEGF) for tissue engineering. Collagen solution was formed into microspheres under a water-in-oil emulsion condition, followed by crosslinking with water-soluble carbodiimide. Various sizes of collagen microspheres in the range of 1-30 mum diameters could be obtained by controlling the surfactant concentration and rotating speed of the emulsified mixture. Particle size proportionally decreased with increasing the rotating speed (1.8 mum per 100 rpm increase in the range of 300-1,200 rpm) and surfactant concentration (3.1 mum per 0.1% increase in the range of 0.1-0.5%). The collagen microspheres showed a slight positive charge of 8.86 and 3.15 mV in phosphate-buffered saline and culture medium, respectively. Release study showed the sustained release of rhVEGF for 4 weeks. Released rhVEGF was able to induce capillary formation of human umbilical vein endothelial cells, indicating the maintenance of rhVEGF bioactivity after release. In conclusion, the results suggest that the collagen microspheres have potential for sustained release of rhVEGF.
Assuntos
Microesferas , Fatores de Crescimento do Endotélio Vascular/farmacologia , Materiais Biocompatíveis , Colágeno , Emulsificantes , Emulsões , Células Endoteliais/efeitos dos fármacos , Humanos , Engenharia Tecidual , Veias Umbilicais/efeitos dos fármacos , Fator A de Crescimento do Endotélio Vascular/farmacologiaAssuntos
Proteínas de Ciclo Celular/genética , Proteínas de Ciclo Celular/farmacologia , Neovascularização de Coroide/tratamento farmacológico , Epitélio Pigmentado da Retina/efeitos dos fármacos , Epitélio Pigmentado da Retina/fisiologia , Animais , Antimutagênicos/farmacologia , Proteínas de Ciclo Celular/metabolismo , Linhagem Celular , Neovascularização de Coroide/genética , Neovascularização de Coroide/patologia , Cobalto/farmacologia , DNA Complementar/farmacologia , Expressão Gênica/fisiologia , Terapia Genética/métodos , Humanos , Hipóxia/tratamento farmacológico , Hipóxia/genética , Hipóxia/patologia , Ratos , Epitélio Pigmentado da Retina/citologia , Fator A de Crescimento do Endotélio Vascular/antagonistas & inibidores , Fator A de Crescimento do Endotélio Vascular/metabolismoRESUMO
A transscleral drug-delivery device, designed for the administration of protein-type drugs, that consists of a drug reservoir covered with a controlled-release membrane was manufactured and tested. The controlled-release membrane is made of photopolymerized polyethylene glycol dimethacrylate (PEGDM) that contains interconnected collagen microparticles (COLs), which are the routes for drug permeation. The results showed that the release of 40-kDa FITC-dextran (FD40) was dependent on the COL concentration, which indicated that FD40 travelled through the membrane-embedded COLs. Additionally, the sustained-release drug formulations, FD40-loaded COLs and FD40-loaded COLs pelletized with PEGDM, fine-tuned the release of FD40. Capsules filled with COLs that contained recombinant human brain-derived neurotrophic factor (rhBDNF) released bioactive rhBDNF in a manner dependent on the membrane COL concentration, as was found for FD40 release. When capsules were sutured onto sclerae of rabbit eyes, FD40 was found to spread to the retinal pigment epithelium. Implantation of the device was easy, and it did not damage the eye tissues. In conclusion, our capsule is easily modified to accommodate different release rates for protein-type drugs by altering the membrane COL composition and/or drug formulation and can be implanted and removed with minor surgery. The device thus has great potential as a conduit for continuous, controlled drug release.