Human mesenchymal stem cell loading as a novel targeting and drug delivery system for stroke.

Andrews, Alexander M (2015) Human mesenchymal stem cell loading as a novel targeting and drug delivery system for stroke. Masters by Research thesis (MSc), Manchester Metropolitan University.

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Abstract

Stroke is the second most common cause of death globally and yet there have been no new developments towards a therapy in the last 25 years. The main issue that surrounds developing therapeutic options for stroke, is the task of overcoming the blood brain barrier (BBB) and delivering an effective treatment to the brain itself. The complications of effectively delivering treatment after stroke are so monumental, that the efficiency of a new therapeutic drug is directly related to the efficiency of the delivery method. Previous methods of drug delivery such as intravenous injection have proved highly ineffective or gravely invasive for the patient. Human mesenchymal stem cells (HMSCs) provide a unique opportunity for developing a novel targeting and drug delivery system, which could effectively deliver therapeutics to specific regions of infarct following stroke. This work aimed to evaluate and quantify the potential of HMSCs to uptake and release various therapeutics, whilst also investigating whether superparamagnetic iron oxide nanoparticles (SPIOs) could be combined with a fixed magnetic field in order to facilitate HMSC migration. In order to investigate the uptake and release of therapeutics, HMSCs were primed with both fibroblast growth factor 2 (FGF-2) and a cyclin-dependent kinase 5 inhibitory peptide (p5). The uptake and release of these potential therapeutics was investigated using developed enzyme-linked immunosorbent assays, fluorescent microscopy and HPLC-MS/MS. The in vitro effects of p5 were investigated using an experimental stroke model and Western blotting. HMSCs were also primed with SPIOs, to determine whether they could be successfully internalized by the cells and if the magnetic properties of the nanoparticles could be also used to facilitate cellular migration. This was completed by using both fluorescent and bright-field microscopy, combined with a transwell migration assay. The data provided shows the potential of HMSCs both as a drug delivery method and for cellular manipulation in vitro. HMSCs have not only been shown to successfully internalize both potential therapeutics and SPIOs but that this could be directly applied to the development of new treatment options following stoke. Furthermore, these results show that the p5 peptide also provides a leading new development for targeted therapy following stroke and that it could pose a useful candidate for future studies within neurodegenerative diseases.