Please use this identifier to cite or link to this item: doi:10.22028/D291-30811
Title: Controlled Release of rAAV Vectors from APMA-Functionalized Contact Lenses for Corneal Gene Therapy
Author(s): Alvarez-Rivera, Fernando
Rey-Rico, Ana
Venkatesan, Jagadeesh K.
Diaz-Gomez, Luis
Cucchiarini, Magali
Concheiro, Angel
Alvarez-Lorenzo, Carmen
Language: English
Title: Pharmaceutics
Issue: 12
Pages: 4
Publisher/Platform: MDPI
Year of Publication: 2020
Free key words: therapeutic contact lens
gene therapy
rAAV vectors
controlled release
corneal diseases
DDC notations: 500 Science
Publikation type: Journal Article
Abstract: As an alternative to eye drops and ocular injections for gene therapy, the aim of this work was to design for the first time hydrogel contact lenses that can act as platforms for the controlled delivery of viral vectors (recombinant adeno-associated virus, rAAV) to the eye in an effective way with improved patient compliance. Hydrogels of hydroxyethyl methacrylate (HEMA) with aminopropyl methacrylamide (APMA) (H1: 40, and H2: 80 mM) or without (Hc: 0 mM) were synthesized, sterilized by steam heat (121 °C, 20 min), and then tested for gene therapy using rAAV vectors to deliver the genes to the cornea. The hydrogels showed adequate light transparency, oxygen permeability, and swelling for use as contact lenses. Loading of viral vectors (rAAV-lacZ, rAAV-RFP, or rAAV-hIGF-I) was carried out at 4 °C to maintain viral vector titer. Release in culture medium was monitored by fluorescence with Cy3-rAAV-lacZ and AAV Titration ELISA. Transduction efficacy was tested through reporter genes lacZ and RFP in human bone marrow derived mesenchymal stem cells (hMSCs). lacZ was detected with X-Gal staining and quantified with Beta-Glo®, and RFP was monitored by fluorescence. The ability of rAAV-hIGF-I-loaded hydrogels to trigger cell proliferation in hMSCs was evaluated by immunohistochemistry. Finally, the ability of rAAV-lacZ-loaded hydrogels to transduce bovine cornea was confirmed through detection with X-Gal staining of β-galactosidase expressed within the tissue.
DOI of the first publication: 10.3390/pharmaceutics12040335
Link to this record: urn:nbn:de:bsz:291--ds-308114
ISSN: 1999-4923
Date of registration: 1-Feb-2021
Description of the related object: Supplementary Material
Related object:
Faculty: M - Medizinische Fakultät
Department: M - Orthopädie
Professorship: M - Prof. Dr. Henning Madry
Collections:SciDok - Der Wissenschaftsserver der Universität des Saarlandes

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