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Transfection of Tenon´s Fibroblasts with Oligonucleotides in Vitro: An Innovative Concept to Influence Wound Modulation Following Glaucoma Surgery
1Luther T. T., 2Welsandt G., 1Jordan J. F., 1Krieglstein G. K., 1Mietz H.,
1Universität zu Köln, Zentrum für Augenheilkunde (Köln)
2Universität zu Köln, Zentrum für Augenheilkunde, Abteilung für Netzhaut- und Glaskörperchirurgie (Köln)
Purpose: Wound modulation following filtering glaucomatous procedures is still of current interest and a field of ongoing research. Clinically, cytotoxic substances are favoured, bearing the risk of severe side-effects. With the concept presented here, a new approach was started: Specific gene sequences are introduced into the target cells. As soon as these cells start to proliferate, the apoptotic mechanism to initiate cell death is started.
Methods: 3T3 fibroblasts and human tenon fibroblasts were plated in 96-well-plates in different densities. After 24 hours a mixture of polycationic lipids (Metafectene) and p53 antisense oligonucleotides were added. After another 24 hour incubation a cristall-violett-essay was performed to determine cell proliferation and survival. Treatment with mitomycin served as control.
Results: Cellular penetration of the liposome-DNA-complex was quantified as high by the fluorescein-label of the oligonucleotide. In p53 antisense oligonucleotide treated cells there was significant growth inhibition compared to cells, which were treated otherwise the same. The inhibition was comparable to effects, which were seen using mitomycin.
Conclusion: The technique presented here is a new approach to influence postoperative cell proliferation after filtering glaucoma surgery. It remains to be seen how effective this new approach may be in in-vivo experiments.