Intravitreal injections of GDNF-loaded biodegradable microspheres are neuroprotective in a rat model of glaucoma.
OBJECTIVE
To evaluate the efficacy of intravitreal injection of GDNF-loaded biodegradable microspheres in promoting the survival of retinal ganglion cells (RGCs) and their axons in a rat model of chronically elevated intraocular pressure (IOP).


METHODS
Chronic elevation of IOP was induced in Brown Norway rats through injection of hypertonic saline (1.9 M) into the episcleral veins. After injection, IOP was measured twice a week in rats using topical anesthesia. Poly DL-lactide-co-glycolide (PLGA) microspheres were fabricated using a modified version of the spontaneous emulsification technique. Two and ten percent of volume solutions of microspheres loaded with glial cell line-derived neurotrophic factor (GDNF) were injected into the vitreous cavity of rats with elevated IOP, with injections of blank microspheres and PBS serving as controls. Histological analysis was used to quantify surviving RGCs and axons and provide comparison among different groups. In addition, the thickness of the retinal inner plexiform layer (IPL) and the level of glial fibrillary acidic protein (GFAP) expression within the retina and optic nerve were quantitatively compared.


RESULTS
IOP was significantly increased in eyes with episcleral vein injection over untreated eyes (p<0.001) but did not show a significant difference among groups that received intravitreal injections of GDNF microspheres, blank microspheres, or PBS (p=0.1852). The duration of IOP elevation in this experiment was eight weeks. Expression of GDNF and its receptors localizes to the adult rat RGCs. Ten percent of the GDNF microsphere treatment significantly increased RGC survival and axon survival (p<0.001), reduced the loss of retinal IPL thickness (p<0.001), and decreased glial cell activation in the retina and optic nerve (p<0.001) compared with blank microspheres and PBS. In addition, GDNF microsphere treatment moderately reduced cupping of the optic nerve head.


CONCLUSIONS
Delivery of GDNF via biodegradable microspheres significantly increased the survival of RGCs and their axons, preserved IPL thickness, and decreased retina and optic nerve glial cell activation in an experimental glaucoma model. This study suggests that GDNF delivered by PLGA microspheres may be useful as a neuroprotective tool in the treatment of glaucomatous optic neuropathy.
Jiang
Caihui
C
Schepens Eye Research Institute, Department of Ophthalmology, Harvard Medical School, Boston, MA 02114, USA.
Moore
Michael J
MJ
Zhang
Xinmei
X
Klassen
Henry
H
Langer
Robert
R
Young
Michael
M
D020341:Absorbable Implants; D000818:Animals; D001369:Axons / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D005901:Glaucoma / Q000473:pathology / Q000503:physiopathology*; D051100:Glial Cell Line-Derived Neurotrophic Factor / Q000008:administration & dosage* / Q000378:metabolism / Q000494:pharmacology; D051097:Glial Cell Line-Derived Neurotrophic Factor Receptors / Q000378:metabolism; D007267:Injections; D007429:Intraocular Pressure / Q000187:drug effects; D008863:Microspheres; D018696:Neuroprotective Agents / Q000008:administration & dosage* / Q000494:pharmacology; D009900:Optic Nerve / Q000187:drug effects / Q000378:metabolism / Q000503:physiopathology*; D051096:Proto-Oncogene Proteins c-ret / Q000378:metabolism; D051381:Rats; D011914:Rats, Inbred BN; D012160:Retina / Q000187:drug effects / Q000378:metabolism / Q000473:pathology / Q000503:physiopathology*; D012165:Retinal Ganglion Cells / Q000187:drug effects / Q000473:pathology; D012462:Saline Solution, Hypertonic / Q000008:administration & dosage / Q000494:pharmacology; D014822:Vitreous Body
C494989:Gfra1 protein, rat; C494992:Gfra2 protein, rat; D051100:Glial Cell Line-Derived Neurotrophic Factor; D051097:Glial Cell Line-Derived Neurotrophic Factor Receptors; D018696:Neuroprotective Agents; D012462:Saline Solution, Hypertonic; D051096:Proto-Oncogene Proteins c-ret; C495001:Ret protein, rat