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N. Singh, P. Jani, J. M. Simonis, R. Kaur, J. Ambati, U. B. Kompella, B. K. Ambati; Albumin Nanoparticles Can Deliver and Express Flt23k Intraceptor for Long Term. Invest. Ophthalmol. Vis. Sci. 2008;49(13):4781. doi: https://doi.org/.
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© ARVO (1962-2015); The Authors (2016-present)
To determine whether albumin nanoparticles can deliver and express Flt23K intraceptor, a recombinant construct of domains 2 to 3 of VEGFR-1 (Flt) coupled with an endoplasmic reticulum retention signal (KDEL), for long term.
FITC conjugated biodegradable albumin nanoparticles encapsulating pCMV.Flt23K and empty pCMV were developed as a lyophilized product that is easily redispersed in an aqueous medium. Nanoparticles were injected into the corneas of BALB/c mice and observed for 4 weeks. Entry of nanoparticles into corneal cells was demonstrated through transmission electron microscopy and confocal imaging. FLt2-3k expression and localization was observed by IP and western blotting
TEM confirmed that nanoparticles were taken up into the corneal keratocyte cytoplasm. FITC-conjugated nanoparticles were expressed in corneal keratocytes and fluorescence could be observed up to 4 weeks. Flt23K was expressed in the corneas for 5 weeks. Organelle fractionation confirmed the localization of Flt23K in the endoplasmic reticulum.
Albumin nanoparticles can deliver and express the FLT2-3K intraceptor for long term into cornea and seem promising for drug delivery. Sustained nonviral gene therapy to the cornea seems viable as a therapeutic approach.
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