Development of Hybrid Nanoplatforms for Theranostic Applications

There is a need of new approaches in which cancer and other diseases are adressed. These new methods must be capable to reduce doses, treatment times, and damage to healty tissues. Furthermore, it is desirable that these systems function as diagnosis and imaging agents, so the evolution of the disease can be visualized in real time and decisions about treatment regimes can be made as soon as possible. The combination of therapeutic and diagnostic/imaging agents in one system creates a new concept called theranostics; hence, a theranostic system can treat the disease and, at the same time, work as a diagnostic and imaging agent. The objective of this thesis has been the synthesis, characterization and evaluation of different multifunctional theranostic nanoplatforms that can be applied in cancer treatment, diagnosis and imaging. The processes used to prepare them have involved low energy consumption and most of the materials were biocompatible. The results obtained from their physicochemical characterization and performance in vitro and in vivo suggest that these nanoplatforms could be used as anticancer treatments with more effective results, allowing lower doses, treatment times, and combined therapies applied locally and simultaneously

Intraarterial route increases the risk of cerebral lesions after mesenchymal cell administration in animal model of ischemia

Mesenchymal stem cells (MSCs) are a promising clinical therapy for ischemic stroke. However, critical parameters, such as the most effective administration route, remain unclear. Intravenous (i.v.) and intraarterial (i.a.) delivery routes have yielded varied outcomes across studies, potentially due to the unknown MSCs distribution. We investigated whether MSCs reached the brain following i.a. or i.v. administration after transient cerebral ischemia in rats, and evaluated the therapeutic effects of both routes. MSCs were labeled with dextran-coated superparamagnetic nanoparticles for magnetic resonance imaging (MRI) cell tracking, transmission electron microscopy and immunohistological analysis. MSCs were found in the brain following i.a. but not i.v. administration. However, the i.a. route increased the risk of cerebral lesions and did not improve functional recovery. The i.v. delivery is safe but MCS do not reach the brain tissue, implying that treatment benefits observed for this route are not attributable to brain MCS engrafting after stroke.This study has been partially supported by grants from Axencia Galega de Innovación (Xunta de Galicia), the Instituto de Salud Carlos III (PI13/00292; PI14/01879), the Spanish Research Network on Cerebrovascular Diseases RETICS-INVICTUS (RD12/0014), Xunta de Galicia (Consellería Educación GRC2014/027), the European Commission program FEDER and Promoting Active Ageing program: Functional Nanostructures For Alzheimer’s Disease At Ultra-Early Stages” (Pana_686009), a Research and Innovation Project, funded within the EU Horizon 2020 Programme”. Furthermore, this study was also co-funded within the POCTEP (Operational Programme for Cross-border Cooperation Spain-Portugal) program (0681_INVENNTA_1_E), co-financed by the ERDF (European Regional Development Fund). T. Sobrino (CP12/03121) and F. Campos (CP14/00154) are recipients of a research contract from Miguel Servet Program of Instituto de Salud Carlos III. Finally, P. Taboada thanks Mineco and Xunta de Galicia for funding through projects MAT2013-40971-R and EM2013-046, respectively. J Trekker is the recipient of an innovation grant from the IWT-VlaanderenS

Tetrahydroxyquinone: A Suitable Coating for Ferrofluids Used in Magnetic Hyperthermia

In this work, tetrahydroxyquinone (THQ) was used for the first time to coat iron oxide nanoparticles (IONPs) and to carry out in vitro experiments in magnetic hyperthermia. Synthesis by co-precipitation resulted in spherical IONPs with a core diameter of 13 ± 3 nm and covered by a 0.5 nm thick coat of THQ, which provided them with a reasonably good zeta potential of ζ = −28 ± 2 mV at pH = 7.3, and thus colloidal stability. The magnetic properties of the THQ-coated IONPs are promising: the low coercive field of Hc = 7 Oe, the high magnetic saturation of Ms = 70.5 emu/g and the low blocking temperature of Tb = 273 K indicate superparamagnetic characteristics at room temperature. Additionally, a high specific absorption rate SAR = 135 W/g (at 300 Oe and 530 kHz) was determined. Cell biological experiments using the human cell line HT-29 evidenced negligible cytotoxicity up to 2 mg/mL. Magnetic hyperthermia (MHT) assays demonstrated fast and reliable heating and reduced the metabolic activity of the cells to 42% upon reaching 42 °C within 15 min. The production of ROS by THQ-coated IONPs could not be detected, which may indicate a reduction in the undesired side effects caused by oxidative stress. Considering these good physicochemical and cell biological properties, this ferrofluid is a promising candidate for the initiation of in vivo experiments for cancer treatment by MHT in murine models

Quantum Dot Labelling of Tepary Bean (Phaseolus acutifolius) Lectins by Microfluidics

Lectins are bioactive proteins with the ability to recognize cell membrane carbohydrates in a specific way. Diverse plant lectins have shown diagnostic and therapeutic potential against cancer, and their cytotoxicity against transformed cells is mediated through the induction of apoptosis. Previous works have determined the cytotoxic activity of a Tepary bean (Phaseolus acutifolius) lectin fraction (TBLF) and its anti-tumorigenic effect on colon cancer. In this work, lectins from the TBLF were additionally purified by ionic-exchange chromatography. Two peaks with agglutination activity were obtained: one of them was named TBL-IE2 and showed a single protein band in two-dimensional electrophoresis; this one was thus selected for coupling to quantum dot (QD) nanoparticles by microfluidics (TBL-IE2-QD). The microfluidic method led to low sample usage, and resulted in homogeneous complexes, whose visualization was achieved using multiphoton and transmission electron microscopy. The average particle size (380 nm) and the average zeta potential (−18.51 mV) were determined. The cytotoxicity of the TBL-IE2 and TBL-IE2-QD was assayed on HT-29 colon cancer cells, showing no differences between them (p ≤ 0.05), where the LC50 values were 1.0 × 10−3 and 1.7 × 10−3 mg/mL, respectively. The microfluidic technique allowed control of the coupling between the QD and the protein, substantially improving the labelling process, providing a rapid and efficient method that enabled the traceability of lectins. Future studies will focus on the potential use of the QD-labelled lectin to recognize tumor tissues