Synthesis and functionalization of inorganic nanoparticles for interactions with bacteria and cells

Diese Arbeit befasst sich mit der Synthese und Charakterisierung von Calciumphosphat und Gold-Nanopartikeln. Es werden die Synthese und kolloidchemische Charakterisierung von Polyelektrolyt-stabilisierten silberdotierten und undotierten Calciumphosphat-Nanopartikeln vorgestellt und ihre biologische Wirkung gegenüber Bakterien und humanen Zellen in vitro diskutiert. Silberdotierte Calciumphosphat-Nanopartikel zeigen bei entsprechenden Silberkonzentrationen vergleichbare toxische Eigenschaften wie Silberacetatlösungen. Es konnte gezeigt werden, dass die biologisch wirksamen, d. h. toxischen Konzentrationen, gegenüber hMSCs, PBMCs, S. aureus und E. coli mit 0,5 bis 5 μg/mL im gleichen Bereich liegen. Darüber hinaus wird die Synthese und kolloidchemische Charakterisierung von unterschiedlich funktionalisierten Gold-Nanopartikeln dargestellt. Verschiedene Strategien zur Funktionalisierung der Gold-Nanopartikel mit Antikörpern werden vorgestellt und die kolloidchemische Charakterisierung der Antikörper-Gold-Nanopartikel-Konjugate wird diskutiert. Es konnte gezeigt werden, dass die rein adsorptiven Wechselwirkungen zwischen Antikörpern und Gold-Nanopartikeln mit Antikörper-Gold-Nanopartikel-Konjugaten durch Kopplung mit EDC und NHS vergleichbar sind

Development of multilayer Hydroxyapatite - Ag/TiN-Ti coatings deposited by radio frequency magnetron sputtering with potential application in the biomedical field

"NOTICE: this is the author's version of a work that was accepted for publication in Surface and Coatings Technology. Changes resulting from the publishing process, such as peer review, editing, corrections, structural formatting, and other quality control mechanisms may not be reflected in this document. Changes may have been made to this work since it was submitted for publication. A definitive version was subsequently published in Surface and Coatings Technology, VOL 377, (2019) DOI 10.1016/j.surfcoat.2019.06.097"[EN] The use of composite coatings is emerging as a great alternative to conventional coatings, allowing the combination of different superficial properties that are widely desired in surgical implants, such as osteointegration and bactericidal character, and cannot be provided by one material alone. In the present investigation the effect of the incorporation of a TiN-Ti intermediate bilayer on the chemical composition, structure, morphology, roughness, residual stresses and adhesion of a multi-layer Hydroxyapatite (HA)-Ag coating deposited on Ti-6Al-4V by magnetron sputtering was evaluated. Additionally, the cytotoxicity of the developed system was evaluated by in vitro tests. According to the results obtained, a decrease in the Ca/P ratio from 1.85 to 1.74 was obtained through the deposition of an HA-Ag system on the intermediate bilayer, and the crystallinity of the developed coating was favored. The multi-layer structure was effectively observed by field emission scanning electron microscopy, where it was possible to identify each of the HA, Ag, TiN and Ti layers. Meanwhile, an increase of 7% in crystallite size, a decrease of 36% in residual stresses and an increase of 32% in adhesion were registered for this composite coating compared to the free intermediate bilayer system. Finally, biological evaluation allowed the non-cytotoxic character of the deposited coatings to be confirmed.We thank the University of Antioquia, the Centro de Investigation, Innovation y Desarrollo de materiales (CIDEMAT) group, the Departamento Administrativo de Ciencia, Tecnologia e Innovation (COLCIENCIAS) for financing the Project 15-1696, the scholarship program of Enlazamundos, PR and JLGR acknowledge financial support from the Spanish Ministry of Economy and Competitiveness (MINECO) through the project MAT2016-76039-C4-1-R (AEI/FEDER, UE) (including the FEDER financial support). CIBER-BBN is an initiative funded by the VI National R&D&I Plan 2008-2011, Iniciativa Ingenio 2010, Consolider Program. 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Natural-based nanocomposites for bone tissue engineering and regenerative medicine: a review

Tissue engineering and regenerative medicine has been providing exciting technologies for the development of functional substitutes aimed to repair and regenerate damaged tissues and organs. Inspired by the hierarchical nature of bone, nanostructured biomaterials are gaining a singular attention for tissue engineering, owing their ability to promote cell adhesion and proliferation, and hence new bone growth, compared with conventional microsized materials. Of particular interest are nanocomposites involving biopolymeric matrices and bioactive nanosized fi llers. Biodegradability, high mechanical strength, and osteointegration and formation of ligamentous tissue are properties required for such materials. Biopolymers are advantageous due to their similarities with extracellular matrices, specifi c degradation rates, and good biological performance. By its turn, calcium phosphates possess favorable osteoconductivity, resorbability, and biocompatibility. Herein, an overview on the available natural polymer/calcium phosphate nanocomposite materials, their design, and properties is presented. Scaffolds, hydrogels, and fi bers as biomimetic strategies for tissue engineering, and processing methodologies are described. The specifi c biological properties of the nanocomposites, as well as their interaction with cells, including the use of bioactive molecules, are highlighted. Nanocomposites in vivo studies using animal models are also reviewed and discussed.  The research leading to this work has received funding from the European Union's Seventh Framework Programme (FP7/2007-2013) under grant agreement no REGPOT-CT2012-316331-POLARIS, and from QREN (ON.2 - NORTE-01-0124-FEDER-000016) cofinanced by North Portugal Regional Operational Program (ON.2 - O Novo Norte), under the National Strategic Reference Framework (NSRF), through the European Regional Development Fund (ERDF)

Uptake of the proteins HTRA1 and HTRA2 by cells mediated by calcium phosphate nanoparticles

The efficient intracellular delivery of (bio)molecules into living cells remains a challenge in biomedicine. Many biomolecules and synthetic drugs are not able to cross the cell membrane, which is a problem if an intracellular mode of action is desired, for example, with a nuclear receptor. Calcium phosphate nanoparticles can serve as carriers for small and large biomolecules as well as for synthetic compounds. The nanoparticles were prepared and colloidally stabilized with either polyethyleneimine (PEI; cationic nanoparticles) or carboxymethyl cellulose (CMC; anionic nanoparticles) and loaded with defined amounts of the fluorescently labelled proteins HTRA1, HTRA2, and BSA. The nanoparticles were purified by ultracentrifugation and characterized by dynamic light scattering and scanning electron microscopy. Various cell types (HeLa, MG-63, THP-1, and hMSC) were incubated with fluorescently labelled proteins alone or with protein-loaded cationic and anionic nanoparticles. The cellular uptake was followed by light and fluorescence microscopy, confocal laser scanning microscopy (CLSM), and flow cytometry. All proteins were readily transported into the cells by cationic calcium phosphate nanoparticles. Notably, only HTRA1 was able to penetrate the cell membrane of MG-63 cells in dissolved form. However, the application of endocytosis inhibitors revealed that the uptake pathway was different for dissolved HTRA1 and HTRA1-loaded nanoparticles