Enhanced oligonucleotide delivery to mouse retinal cells using iontophoresis.

Details

Serval ID
serval:BIB_B21384C2850B
Type
Article: article from journal or magazin.
Collection
Publications
Institution
Title
Enhanced oligonucleotide delivery to mouse retinal cells using iontophoresis.
Journal
Molecular Vision
Author(s)
Andrieu-Soler C., Doat M., Halhal M., Keller N., Jonet L., BenEzra D., Behar-Cohen F.
ISSN
1090-0535 (Electronic)
ISSN-L
1090-0535
Publication state
Published
Issued date
2006
Peer-reviewed
Oui
Volume
12
Pages
1098-1107
Language
english
Notes
Publication types: Journal Article ; Research Support, Non-U.S. Gov'tPublication Status: epublish
Abstract
PURPOSE: To study the combination of oligodeoxynucleotides (ODNs) intravitreous injection and saline transpalpebral iontophoresis on the delivery of ODNs to photoreceptors in the newborn rd1/rd1 mice.
METHODS: Cathodal or anodal transpalpebral iontophoresis (1.43 mA/cm(2) for 5 min) was applied to eyes of postnatal day 7 (PN7) rd1/rd1 mice immediately before the intravitreous injection of ODNs. The effect of cathodal iontophoresis after ODNs injection was also evaluated. The influence of current intensity (0.5, 1.5, and 2.5 mA) was assayed with cathodal iontophoresis performed prior to ODNs injection. The duration of current-induced facilitation of ODNs delivery to photoreceptors was evaluated for 6 h following iontophoresis. One group of control eyes received cathodal iontophoresis prior to the intravitreous injection of phosphate buffered saline (PBS) or hexachlorofluorescein (Hex). The second control group received ODN or Hex intravitreous injection without iontophoresis. The penetration of fluorescent ODNs in the outer nuclear layer (ONL) was quantified by image analysis of the ONL fluorescence intensity on cryosection microphotographs. Integrity of ODN was assessed using acrylamide gel migration after its extraction from the retina of treated mice. The integrity of retinal structure, 1 and 24 h after iontophoresis, was analyzed using light and electron microscopy.
RESULTS: Transpalpebral anodal or cathodal saline iontophoresis enhanced the penetration of ODNs in all retinal layers. Cathodal iontophoresis was more efficient than anodal iontophoresis in enhancing the tissue penetration of the injected ODN. Photoreceptor delivery of ODN was significantly higher when cathodal saline transpalpebral iontophoresis was applied prior than after the injection. The extent of enhanced tissue penetration decreased in parallel to the increased interval between iontophoresis application and the intravitreous injection. Current of 1.5 mA was safe and optimal for the delivery of ODNs to the ONL. One hour after iontophoresis followed by injection, ODN extracted from the retina of treated eyes remained intact. Histology and electron microscopy observations demonstrated that iontophoresis using the optimal parameters did not induce any permanent tissue alterations or structure damage.
CONCLUSIONS: Saline transpalpebral iontophoresis facilitates the penetration of injected ODNs in photoreceptors for at least 3 h. This method may be considered for photoreceptor targeted gene therapy.
Keywords
Animals, Animals, Newborn, Injections, Iontophoresis, Mice, Mice, Inbred C3H, Microscopy, Electron, Oligonucleotides/administration & dosage, Oligonucleotides/pharmacokinetics, Photoreceptor Cells, Retina/metabolism, Retina/pathology, Retinal Degeneration/genetics, Retinal Degeneration/metabolism, Tissue Distribution, Vitreous Body
Pubmed
Web of science
Create date
26/09/2013 15:06
Last modification date
20/08/2019 15:20
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