Sustained subconjunctival protein delivery using a thermosetting gel delivery system.
OBJECTIVE
An effective treatment modality for posterior eye diseases would provide prolonged delivery of therapeutic agents, including macromolecules, to eye tissues using a safe and minimally invasive method. The goal of this study was to assess the ability of a thermosetting gel to deliver a fluorescently labeled protein, Alexa 647 ovalbumin, to the choroid and retina of rats following a single subconjunctival injection of the gel. Additional experiments were performed to compare in vitro to in vivo ovalbumin release rates from the gel.


METHODS
The ovalbumin content of the eye tissues was monitored by spectrophotometric assays of tissue extracts of Alexa 647 ovalbumin from dissected sclera, choroid, and retina at time points ranging from 2 h to 14 days. At the same time points, fluorescence microscopy images of tissue samples were also obtained. Measurement of intact ovalbumin was verified by LDS-PAGE analysis of the tissue extract solutions. In vitro release of Alexa 488 ovalbumin into 37 degrees C PBS solutions from ovalbumin-loaded gel pellets was also monitored over time by spectrophotometric assay. In vivo ovalbumin release rates were determined by measurement of residual ovalbumin extracted from gel pellets removed from rat eyes at various time intervals.


RESULTS
Our results indicate that ovalbumin concentrations can be maintained at measurable levels in the sclera, choroid, and retina of rats for up to 14 days using the thermosetting gel delivery system. The concentration of ovalbumin exhibited a gradient that decreased from sclera to choroid and to retina. The in vitro release rate profiles were similar to the in vivo release profiles.


CONCLUSIONS
Our findings suggest that the thermosetting gel system may be a feasible method for safe and convenient sustained delivery of proteins to choroidal and retinal tissue in the posterior segments of the eye.
Laboratory of Bioengineering and Physical Sciences, National Institute of Biomedical Imaging and Bioengineering, National Eye Institute, National Institutes of Health, Bethesda, Maryland 20892-5766, USA.
Erin R
ER
Rieke
Juan
J
Amaral
S Patricia
SP
Becerra
Robert J
RJ
Lutz
D000818:Animals; D002829:Choroid / Q000378:metabolism*; D003692:Delayed-Action Preparations; D016503:Drug Delivery Systems; D005240:Feasibility Studies; D005782:Gels; D008297:Male; D009994:Osmolar Concentration; D010047:Ovalbumin / Q000008:administration & dosage* / Q000493:pharmacokinetics*; D051381:Rats; D011914:Rats, Inbred BN; D012160:Retina / Q000378:metabolism*; D012590:Sclera / Q000378:metabolism*; D013696:Temperature
D003692:Delayed-Action Preparations; D005782:Gels; D010047:Ovalbumin