Low molecular weight hyaluronan shielding of DNA/PEI polyplexes facilitates CD44 receptor mediated uptake in human corneal epithelial cells.
OBJECTIVE
It was the aim of this study to prepare purified DNA/PEI polyplexes, which are coated with hyaluronan to facilitate CD44 receptor mediated uptake of the DNA/PEI polyplex and to reduce unspecific interactions of the complex with negatively charged extracellular matrix components on the ocular surface.


METHODS
Hyaluronans of different molecular weights (<10 kDa, 10-30 kDa and 30-50 kDa) were isolated after enzymatic degradation of high molecular weight hyaluronan via ultrafiltration by centrifugation. The influence of the different hyaluronans used for coating on the stability and transfection efficiency of the complexes was evaluated in vitro. Transfection and uptake studies were performed in human corneal epithelial (HCE) cells. CD44 receptor expression of this cell model was evaluated by immunohistochemistry.


RESULTS
Coating of purified DNA/PEI polyplexes with low molecular weight hyaluronan (<10 kDa) facilitated receptor-mediated uptake via the CD44 receptor in HCE cells, increased complex stability in vitro, and effectively shielded the positive surface charges of the polyplex without decreasing its transfection efficiency. Higher molecular weights and larger amounts of hyaluronan in the complexes resulted in lesser improvements in the stability and transfection efficacy of the complexes.


CONCLUSIONS
Coating of polyplexes with low molecular weight hyaluronan is a promising strategy for gene delivery to the ocular surface, where CD44 receptor mediated uptake decreased cytotoxicity and reduced non-specific interactions with the negatively charged extracellular matrix components are considered beneficial for increased transfection efficiency of non-viral vectors.
Hornof
Margit
M
Drug Discovery and Development Technology Center DDTC, University of Helsinki, Finland.
de la Fuente
Maria
M
Hallikainen
Marjut
M
Tammi
Raija H
RH
Urtti
Arto
A
D001386:Azides / Q000378:metabolism; D002478:Cells, Cultured; D004247:DNA / Q000378:metabolism*; D019573:Epithelium, Corneal / Q000166:cytology* / Q000378:metabolism*; D005374:Filtration; D005434:Flow Cytometry; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006801:Humans; D018960:Hyaluronan Receptors / Q000378:metabolism*; D006820:Hyaluronic Acid / Q000378:metabolism*; D008970:Molecular Weight; D010316:Particle Size; D010957:Plasmids / Q000378:metabolism; D011094:Polyethyleneimine / Q000378:metabolism*; D021381:Protein Transport; D014162:Transfection; D019076:Transgenes; D001616:beta-Galactosidase / Q000378:metabolism
D001386:Azides; D018960:Hyaluronan Receptors; D049452:Green Fluorescent Proteins; C014535:8-azidoethidium; D011094:Polyethyleneimine; D006820:Hyaluronic Acid; D004247:DNA; D001616:beta-Galactosidase