Nanoparticle‐mediated Transfer of Therapeutics to Corneal Cells

Abstract

AbstractPurpose Calcium phosphate nanoparticles (CaP‐NPs) are biodegradable and biocompatible as they dissolve in the cell in calcium and phosphate. Therefore they are an ideal tool for transfection with DNA or RNA. It has been demonstrated that the impact on intracellular levels of Ca2+ is low and therefore does not affect cell vitality. In this study, we determine the most efficient type of CaP‐NPs for transfer of DNA into in corneal endothelial cells (EC).Methods CaP‐NPs with triple‐shell pcDNA3‐EGFP in different concentrations or coated by dispersions of polyethylenimine (PEI) were manufactured. Polyfect®‐pcDNA3‐EGFP served as positive control. Human and murine EC (suspensions and donor tissue) were transfected using various concentrations of CaP‐NPs and different times of transfection. Transfection efficacy was determined by measuring EGFP‐expression (flow cytometry, fluorescence microscopy). To evaluate toxicity, apoptosis was studied by TUNEL and propidium iodide staining.Results The transfection efficacy of triple shell CaP‐NPs (EGFP expression up to 50% after transfection with triple shell CaP‐NPs) was significantly increased after coating with PEI. Cell viability in corneal tissue was not significantly impaired after treatment. As EC possess minimal proliferative capacity, there was no significant change of EGFP expression.Conclusion CaP‐NPs seem to be an alternative to viral vectors and safe for application in patients. Therefore CaP‐NP may be a new and suitable tool for DNA transfer into EC. Further studies are necessary to carefully evaluate functional application and biosafety.