Aplicación de la espectroscopía de infrarrojo a la identificación y la cuantificación de diferentes compuestos

Memoria ID-0147. Ayudas de la Universidad de Salamanca para la innovación docente, curso 2015-2016

Determinación de la Capacidad Térmica y el Calor Específico

Memoria ID-0303. Ayudas de la Universidad de Salamanca para la innovación docente, curso 2014-2015

Multiparametric analysis of anti-proliferative and apoptotic effects of gold nanoprisms on mouse and human primary and transformed cells, biodistribution and toxicity in vivo

Background: The special physicochemical properties of gold nanoprisms make them very useful for biomedical applications including biosensing and cancer therapy. However, it is not clear how gold nanoprisms may affect cellular physiology including viability and other critical functions. We report a multiparametric investigation on the impact of gold-nanoprisms on mice and human, transformed and primary cells as well as tissue distribution and toxicity in vivo after parental injection. Methods: Cellular uptake of the gold-nanoprisms (NPRs) and the most crucial parameters of cell fitness such as generation of reactive oxygen species (ROS), mitochondria membrane potential, cell morphology and apoptosis were systematically assayed in cells. Organ distribution and toxicity including inflammatory response were analysed in vivo in mice at 3 days or 4 months after parental administration. Results: Internalized gold-nanoprisms have a significant impact in cell morphology, mitochondrial function and ROS production, which however do not affect the potential of cells to proliferate and form colonies. In vivo NPRs were only detected in spleen and liver at 3 days and 4 months after administration, which correlated with some changes in tissue architecture. However, the main serum biochemical markers of organ damage and inflammation (TNFa and IFN¿) remained unaltered even after 4 months. In addition, animals did not show any macroscopic sign of toxicity and remained healthy during all the study period. Conclusion: Our data indicate that these gold-nanoprisms are neither cytotoxic nor cytostatic in transformed and primary cells, and suggest that extensive parameters should be analysed in different cell types to draw useful conclusions on nanomaterials safety. Moreover, although there is a tendency for the NPRs to accumulate in liver and spleen, there is no observable negative impact on animal health

Sterilization matters: Consequences of different sterilization techniques on gold nanoparticles

Nanoparticles (NPs) can offer many advantages over traditional drug design and delivery, as well as toward medical diagnostics. As with any medical device or pharmaceutical drug intended to be used for in vivo biomedical applications, NPs must be sterile. However, very little is known regarding the effect of sterilization methods on the intrinsic properties and stability of NPs. Herein a detailed analysis of physicochemical properties of two types of AuNPs upon sterilization by means of five different techniques is reported. In addition, cell viability and production of reactive oxygen species are studied. The results indicate that sterilization by ethylene oxide seems to be the most appropriate technique for both types of NPs. It is concluded that it is crucial to test several methods in order to establish the specific type of sterilization to be performed for each particular NP.The authors acknowledge financial support from the Xunta de Galicia (PGIDIT06TMT31402PR), SUDOE (IMMUNONET-SOE1/1P1/E014), and Spanish Ministry of Science and Innovation (Consolider Ingenio 2010, CSD2006-12, NANOBIOMED). Ángela França was supported with a Leonardo da Vinci Fellowship. Jesús Martinez de la Fuente thanks ARAID for financial support.Peer reviewe

The Bluegreen Algae (AFA) Consumption over 48 h Increases the Total Number of Peripheral CD34+ Cells in Healthy Patients: Effect of Short-Term and Long-Term Nutritional Supplementation (Curcumin/AFA) on CD34+ Levels (Blood)

Several active principles from plants could trigger the release of stem cells from the bone marrow. Stem cell mobilizers have shown side effects in patients. Thus, the purpose of this paper is to find the natural products from plants (curcuminoids, glycosinolate of sulforaphane, AFA bluegreen algae), which could be potential stem mobilizes without adverse side effects. The antioxidant curcumin [1,7-bis(4-hydroxy-3-methoxyphenyl)-1,6-heptadiene-2,5-dione], glycosinolate of sulforaphane (broccoli) or AFA (Aphanizomenon flos) extract promote beneficial effects in patients. The number of circulating stem cells were monitored by HSC marker-CD34 by flow cytometry in peripheral blood from healthy subjects. CD34 is a hematological stem cells (HSC) marker. A double-blind study was conducted in 22 healthy subjects. We have evaluated whether short-term AFA—Aphanizomenon flos aquae—algae or curcuminoids consumption (powder or liquid formulation) over 48 consecutive hours could increase the total number of peripheral CD34+ blood cells (n = 22, n = 5 subjects/group). The total number of circulating CD34+ cells were quantified after short-term and long-term nutritional supplementation; their levels were compared with their own basal levels (n = 5/group, controls: before taking any supplement) or placebo-treated patients (n = 7); their average age was 54 years old. We also evaluated whether long-term nutritional supplementation with several nutraceuticals could enhance HSC mobilization by increasing the total number of peripheral CD-34+ cell after seven or 38 consecutive days of administration (n = 5, with seven placebo-treated patients). The long-term administration take place with these doses/day [curcuminoids: 2000 mg/day, equivalent to 120 mg of curcuminoids/day), glycosinolate of sulforaphane (66 mg/day), plus AFA Algae bluegreen extract (400 mg/day)]. On the last day (10 a.m.) of treatment, blood samples were collected six hours after taking these supplements; the average age was 54 years old. Notably, the blue green AFA algae extract consumption over 48 h enhances HSC mobilization by increasing the total number of peripheral CD34+ cells. The long-term administration with curcuminoids, glycosinolate of sulforaphane, and AFA bluegreen algae extract also increased the total number of CD34-HSC cells after seven or 38 days of consecutive of administration in healthy subjects

Adult Mesenchymal Stem Cells from Oral Cavity and Surrounding Areas: Types and Biomedical Applications

Adult mesenchymal stem cells are those obtained from the conformation of dental structures (DMSC), such as deciduous and permanent teeth and other surrounding tissues. Background: The self-renewal and differentiation capacities of these adult stem cells allow for great clinical potential. Because DMSC are cells of ectomesenchymal origin, they reveal a high capacity for complete regeneration of dental pulp, periodontal tissue, and other biomedical applications; their differentiation into other types of cells promotes repair in muscle tissue, cardiac, pancreatic, nervous, bone, cartilage, skin, and corneal tissues, among others, with a high predictability of success. Therefore, stem and progenitor cells, with their exosomes of dental origin and surrounding areas in the oral cavity due to their plasticity, are considered a fundamental pillar in medicine and regenerative dentistry. Tissue engineering (MSCs, scaffolds, and bioactive molecules) sustains and induces its multipotent and immunomodulatory effects. It is of vital importance to guarantee the safety and efficacy of the procedures designed for patients, and for this purpose, more clinical trials are needed to increase the efficacy of several pathologies. Conclusion: From a bioethical and transcendental anthropological point of view, the human person as a unique being facilitates better clinical and personalized therapy, given the higher prevalence of dental and chronic systemic diseases

Engineering biofunctional magnetic nanoparticles for biotechnological applications

10 páginas, 6 figuras, 1 esquema.Synthesis and characterization of magnetic nanoparticles with excellent size control are showed here. Their functionalization using an amphiphilic polymer is also described. This strategy allows the stabilization of magnetic nanoparticles in aqueous solvents and in addition, the polymer shell serves as a platform to incorporate relevant biomolecules, such as poly(ethylene glycol) and a number of carbohydrates. Nanoparticles functionalized with carbohydrates show the ability to avoid unspecific interactions between proteins present in the working medium and the nanoparticles, so can be used as an alternative to poly(ethylene glycol) molecules. Results confirm these nanoparticles as excellent contrast agents for magnetic resonance imaging. Changes in the spin-spin transversal relaxation times of the surrounding water protons due to nanoparticle aggregation demonstrates the bioactivity of these nanoparticles functionalized with carbohydrates. To finish with, nanoparticle toxicity is evaluated by means of MTT assay. The obtained results clearly indicate that these nanoparticles are excellent candidates for their further application in nanomedicine or nanobiotechnology. © 2010 The Royal Society of Chemistry.This work was supported by institutional funding from MITYC (CTQ2008-03739/PPQ) (Spain), Community of Madrid (Grant S-BIO2006-170) and NANOBIOMED-Consolider. JMF thanks ARAID for financial support.Peer Reviewe

Functionalized Fe3O4@Au superparamagnetic nanoparticles: in vitro bioactivity

The interaction of nanoparticles with cells has been a focus of interest during the past decade. We report the fabrication and characterization of hydrosoluble Fe3O4@Au nanoparticles functionalized with biocompatible and fluorescent molecules and their interaction with cell cultures by visualizing them with confocal microscopy. Gold covered iron oxide nanoparticles were synthesized by reducing metal salts in the presence of oleylamine and oleic acid. The functionalization of these particles with an amphiphilic polymer provides a water soluble corona as well as the possibility to incorporate different molecules relevant for bio-applications such as poly(ethylene glycol), glucose or a cadaverine derived dye. The particle size, and the presence of polymer layers and conjugated molecules were characterized and confirmed by transmission electron microscopy, thermogravimetric measurements and infrared spectroscopy. A complete magnetic study was performed, showing that gold provides an optimum coating, which enhances the superparamagnetic behaviour observed above 10–15 K in this kind of nanoparticle. The interaction with cells and the cytotoxicity of the Fe3O4@Au preparations were determined upon incubation with the HeLa cell line. These nanoparticles showed no cytotoxicity when evaluated by the MTT assay and it was demonstrated that nanoparticles clearly interacted with the cells, showing a higher level of accumulation in the cells for glucose conjugated nanoparticles.This work was supported by institutional funding from the Ministerio de Educacion y Ciencia and Basque Government under project Nos MAT2010-19942, SAIOTEK SP-10UN64, GIC-IT-382-07 and CTQ2008-03739/PPQ and ERC-Starting Grant 239931-NANOPUZZLE projects. J Salado thanks the Basque Government for a postdoctoral fellowship. J M de la Fuente thanks ARAID for financial support.Peer reviewe

MOESM1 of Nanoparticles engineered to bind cellular motors for efficient delivery

Additional file 1: Figure S1. UV/Vis spectra of “bare” NPs (Au@tiopronin) before the modification, and peptide, PEG or TAMRA-CAD modified NPs. a) DBP (red line, Au@tiopronin-DynPro), IntCT (cyan line, Au@tiopronin-IntCt) and TAMRA (blue line, Au@tiopronin-IntCt-TAMRA-CAD); b) DBP-PEG (red line, Au@tiopronin-DynPro/PEG), IntCT-PEG (cyan line, Au@tiopronin-IntCt/PEG) and TAMRA-PEG (blue line, Au@tiopronin-IntCt-TAMRA-CAD/PEG). Figure S2. Fluorescence spectra of “bare” NPs (Au@tiopronin) before the modification, and peptide, PEG or TAMRA-CAD modified NPs. a) DBP (red line, Au@tiopronin-DynPro), IntCT (cyan line, Au@tiopronin-IntCt) and TAMRA (blue line, Au@tiopronin-IntCt-TAMRA-CAD); b) DBP-PEG (red line, Au@tiopronin-DynPro/PEG), IntCT-PEG (cyan line, Au@tiopronin-IntCt/PEG) and TAMRA-PEG (blue line, Au@tiopronin-IntCt-TAMRA-CAD/PEG). Figure S3. ζ-potential bar diagram of “bare” NPs (Au@tiopronin) before the modification, and modified peptide or TAMRA-CAD modified NPs: DBP (Au@DynPro), IntCT (Au@IntCt), TAMRA (Au@TAMRA-CAD), DBP-PEG (Au@DynPro/PEG), IntCT-PEG (Au@IntCt/PEG) and TAMRA-PEG (Au@TAMRA-CAD/PEG). Table S1. ζ-potential values of “bare” NPs (Au@tiopronin) before the modification, and peptide, PEG or TAMRA-CAD modified NPs: DBP (Au@DynPro), IntCT (Au@IntCt), TAMRA (Au@TAMRA-CAD), DBP-PEG (Au@DynPro/PEG), IntCT-PEG (Au@IntCt/PEG) and TAMRA-PEG (Au@TAMRA-CAD/PEG). Figure S4. The figure shows the cellular uptake (according to intracellular MFI) of Au@DynPro, Au@DynPro-PEG, compared to nanoparticles modified with internal control peptide (IntCt), Au@IntCt and Au@IntCt-PEG. Mean fluorescence intensity (MFI) of modified NPs after 1 h incubation. Figure S5. This figure shows the absence of cytotoxic effect of the NPs in Vero cells. Cell viability (a) and cell proliferation (b) were analyzed after incubation of cells with increasing concentrations of the different Au@DBP that exceeded those used in this study. A significant decrease in cell counts or cell proliferation was not observed