2018-03, Boehringer, Stephan, Ruzgys, Paulius, Tamò, Luca, Satkauskas, Saulius, Geiser, Thomas, Gazdhar, Amiq, Hradetzky, David

A challenge for gene therapy is absence of safe and efficient local delivery of therapeutic genetic material. An efficient and reproducible physical method of electrospray for localized and targeted gene delivery is presented. Electrospray works on the principle of coulombs repulsion, under influence of electric field the liquid carrying genetic material is dispersed into micro droplets and is accelerated towards the targeted tissue, acting as a counter electrode. The accelerated droplets penetrate the targeted cells thus facilitating the transfer of genetic material into the cell. The work described here presents the principle of electrospray for gene delivery, the basic instrument design, and the various optimized parameters to enhance gene transfer in vitro. We estimate a transfection efficiency of up to 60% was achieved. We describe an efficient gene transfer method and a potential electrospray-mediated gene transfer mechanism.

2016, Hradetzky, David, Boehringer, Stephan, Ruzgys, Paulius, Satkauskas, Saulius, Gazdhar, Amiq, Geiser, Thomas

2012, Hradetzky, David, Boehringer, Stephan, Geiser, Thomas, Gazdhar, Amiq

Idiopathic pulmonary fibrosis (IPF) is a devastating disease affecting the distal lung, due to failure of the alveolar epithelium to heal after micro-injuries, leading to inefficient gas exchange and resulting in death. Therapeutic options are very limited. A new therapeutic approach based on gene therapy restores the self-healing process within the lung in the experimental setup. A basic requirement of this therapy is the successful transduction of genes into the alveolar epithelium in the distal part of the lung, for which a new therapeutic instrument is required. In this paper we present the concept and first experimental results of a device which uses an electrical field to accelerate the charged droplets of plasmid suspension toward the tissue and which overcomes cell membrane with its impact energy. The aim is to develop a therapeutic device capable of being integrated into minimally invasive procedures such as bronchoscopy.

2018, Ruzgys, Paulius, Boehringer, Stephan, Tamò, Luca, Šatkauskas, Saulius, Geiser, Thomas, Gazdhar, Amiq, Hradetzky, David

Electrospray is a process based on applied high electric voltage on liquid, leading to the creation and acceleration of small sized droplets. Electrosprayed suspension with exogenous molecules on cells can trigger intracellular delivery. Here, the application study of anticancer drug delivery via electrospray as an antitumor treatment is presented in vitro and in vivo. In vitro experiments were performed on alveolar epithelial cells (A549). Cell viability was performed with flow cytometry assay. Apoptosis/ necrosis was measured using annexin V/PI kit. In vivo experiments were done with C57BL/6J mice. Lewis lung carcinoma (LLC) cell lines were injected intradermally to induce tumor. Electrospray was performed on tumor by cutting the skin. Tumor size was measured with caliper. Results show triggered PI transfer to cells after electrospray process. Moreover, obtained results in vitro showed successful cisplatin and methotrexate intracellular delivery, resulting in around 90 % of cell viability loss with methotrexate and 70 % of cell viability loss with cisplatin. In vivo experiments reveled a tumor decrease by around 2.5 times in 7 days after electrospray treatment at first and third day with both anticancer drugs. Here we show that electrospray method can be applied to local anticancer drug delivery to cells and tissues. Therefore, such method might be adapted as a clinical anticancer therapy in the future.

2015, Ruzgys, Paulius, Boehringer, Stephan, Šatkauskas, Saulius, Hradetzky, David, Gazdhar, Amiq, Geiser, Thomas

2018, Hradetzky, David, Boehringer, Stephan, Ruzgys, Paulius, Šatkauskas, Saulius, Geiser, Thomas, Gazdhar, Amiq

Electrospray is a process based on creation and acceleration of small sized droplets based on electrostatic repulsion. Spraying plasmid containing liquids this process may be used to transfer genes into cells. Within this paper we report on a method for accessing and evaluating the spray modalities using high speed imaging system with a post processing of image data to obtain estimated volume and velocity of emerging droplets first. Second we investigate on the impact of different media on the spray modalities. Third we evaluate the impact of the spray on cell viability and on transfection efficiency of an eGFP plasmid as reporter gene obtained in an in vitro setup on alveolar epithelial like cells (A549).

2013, Fiave, Prosper Agbesi, Boehringer, Stephan, Gazdhar, Amiq, Geiser, Thomas, Hradetzky, David

Electric field accelerated plasmid droplets to permeate tissues has been demonstrated on cell culture and ex-vivo lung tissue of rats using enhanced green fluorescence protein reporter gene for transfection. This opens a window of opportunity to design an electrospray instrument usable within the working channel of bronchoscope to noninvasively treat idiopathic pulmonary fibrosis (IPF). We elaborate a concept and realized a device usable in the working channel of a flexible bronchoscope taking into account restrictions within typical bronchoscopy procedure.