Small upconverting fluorescent nanoparticles for biomedical applications

Fluorescent labels have been widely used for biological applications, primarily in imaging and assays. Traditional fluorophores such as fluorescent dyes are mainly based on downconversion fluorescence, which have several drawbacks such as photobleaching, high background noise from autofluorescence, and considerable photodamage to biological materials. Upconverting fluorescent nanoparticles emit detectable photons of higher energy in the near-infrared (NIR) or visible range upon irradiation with an NIR light in a process termed 'upconversion.' They overcome some of the disadvantages faced by conventional downconversion labels, thus making them an ideal fluorescent label for biological applications. This review looks at the development of these particles, critically examines the reported applications, and discusses their future in biomedicine.     

生物医用磁性纳米颗粒的制备与表面改性

磁性纳米颗粒目前是生物医用纳米材料领域异常活跃的方向之一.不同方法制备的磁性纳米颗粒经不同聚合物或分子表面改性后具有多方面的生物医学应用.本文综合评述了磁性纳米颗粒的制备方法,如共沉淀法、溶胶-凝胶法、微乳剂法等;总结了磁性纳米颗粒表面改性技术,包括改性物质与改性方法;概括了磁性纳米颗粒在生物医学领域的应用,主要涉及磁靶向制剂、细胞分离、肿瘤细胞的过热治疗、MR I衬度增强剂四方面.磁性纳米颗粒还有很大的发展空间和广阔的应用前景.     

Mesoporous silica nanoparticles prepared by different methods for biomedical applications: Comparative study

In the current investigation, mesoporous silica nanoparticles were obtained by various techniques, namely sol–gel (S1), micro‐emulsion (S2) and hydrothermal synthesis (S3). The effect of those methods on the final features of the obtained mesoporous silica nanoparticles was studied. The obtained nanoparticles were investigated by TEM, BET surface area, Zetasizer, XRD and FTIR. The preparation method effect was evaluated on the drug release behaviour using doxycycline hyclate as a model drug. In addition, the degree of their compatibility against Saos‐2 cell line was also determined. The morphology and microstructure of silica nanoparticles were found to be dependent on the utilised method. Those techniques produced particles with particle sizes of 50, 30–20 and 15 nm and also surface areas of 111.04, 164 and 538.72 m²/g, respectively, for S1, S2 and S3. However, different preparation methods showed no remarkable changes for the physical and chemical integrities. The drug release test showed faster release from S2 compared with S1 and S3, which make them more applicable in cases require large doses for short periods. However, the release behaviour of S3 was satisfied for treatments which require long period with relatively highest release rate. The preparation method influenced the cell viability as S1 and S2 showed acceptable cell cytotoxicity compared with S3.     

Highly fluorescent dye-doped silica nanoparticles increase flow cytometry sensitivity for cancer cell monitoring

Early and accurate diagnosis and treatment of cancer depend on rapid, sensitive, and selective detection of tumor cells. Current diagnosis of cancers, especially leukemia, relies on histology and flow cytometry using single dye-labeled antibodies. However, this combination may not lead to high signal output, which can hinder detection, especially when the probes have relatively weak affinities or when the receptor is expressed in a low concentration on the target cell surface. To solve these problems, we have developed a novel method for sensitive and rapid detection of cancer cells using dye-doped silica nanoparticles (NPs) which increases detection sensitivity in flow cytometry analyses between 10- and 100-fold compared to standard methods. Our NPs are ∼60 nm in size and can encapsulate thousands of individual dye molecules within their matrix. We have extensively investigated surface modification strategies in order to make the NPs suitable for selective detection of cancer cells using flow cytometry. The NPs are functionalized with polyethylene glycol (PEG) to prevent nonspecific interactions and with neutravidin to allow universal binding with biotinylated molecules. By virtue of their reliable and selective detection of target cancer cells, these NPs have demonstrated their promising usefulness in conventional flow cytometry. Moreover, they have shown low background signal, high signal enhancement, and efficient functionalization, either with antibody- or aptamer-targeting moieties. Electronic Supplementary Material  Supplementary material is available for this article at and is accessible for authorized users.     

Preparation and biomedical applications of core-shell silica/magnetic nanoparticle composites

Core-shell structured silica/magnetic nanoparticle composites have recently been subjected to extensive research since the shells could offer protection to the cores and introduce new properties to the hybrid structures, which endue them with great application potentials in various fields. Several approaches have been studied for the synthesis of SiO2 coated on magnetic nanoparticles. These approaches include St?ber process, microemulsion, sodium silicate and tetraethoxysilane hydrolysis, aerosol pyrolysis, layer-by-layer strategy, polymer-templating and sonochemical deposition. This review is focused on describing state-of-the-art synthetic routes and methods for the preparation of silica/magnetic nanoparticle composites. Furthermore, we also introduce main applications of these nanoparticle composites in biomedical scopes and address some challenges in the synthesis of high-quality magnetic nanoparticles.     

Structural and morphological investigation of magnetic nanoparticles based on iron oxides for biomedical applications

The present work reports the synthesis, characterization and properties of magnetic iron oxide nanoparticles for biomedical applications, correlating the nanoscale tunabilities in terms of size, structure, and magnetism. Magnetic nanoparticles in different conditions were prepared through thermal decomposition of Fe(acac)₃ in the presence of 1,2 hexadecanodiol (reducing agent) and oleic acid and oleylamine (ligands) in a hot organic solvent. The 2,3-dimercaptosuccinic acid (DMSA) was exchanged onto the nanocrystal surface making the particles stable in water. Nanoparticles were characterized by X-ray diffraction (XRD) measurements, small angle X-ray scattering (SAXS) and transmission electron microscopy (TEM). Preliminary tests of incorporation of these nanoparticles in cells and their magnetic resonance image (MRI) were also carried out. The magnetization characterizations were made by isothermal magnetic measurements.     

Hydrogel-magnetic nanoparticles with immobilized l-asparaginase for biomedical applications

The association of magnetic nanoparticles, which could be controlled by a magnetic field and have dimensions which facilitate their penetration in cells/tissues, with hydrogel type biopolymeric shells confer them compatibility and the capacity to retain and deliver bioactive substances. The main objective of this work is the development of a new system based on a biocompatible polymer with organic–inorganic structure capable of vectoring support for biologic active agents (l-asparaginase, e.g.). Characterization of size and morphology of the hydrogel-magnetic nanoparticles with entrapped l-asparaginase was made using Dynamic Light Scattering method, Transmission Electron Microscopy and Confocal Microscopy. The structure of magnetic nanoparticles coated with hydrogel was characterized by Fourier Transformed Infrared Spectroscopy. The cytotoxicity of nanoparticles was evaluated and also the interactions with microorganisms. We obtained hydrogel-magnetic nanoparticles with l-asparaginase entrapped, with sizes below 30 nm in dried stage, capable to penetrate the cells and tissues.     

Highly selective and sensitive magnetic silica nanoparticles based fluorescent sensor for detection of Zn ions

In this work, quinoline group modified multifunctional silica nanoparticles having high magnetization and excellent Zn²⁺ selectivity have been successfully prepared. These multifunctional nanoparticles were characterized by high resolution transmission electron microscopy (HRTEM), Fourier transform infrared spectroscopy (FT-IR), thermal gravimetric analysis (TGA), X-ray powder diffraction (XRD) and vibrating sample magnetometer (VSM). The characterization data indicated that the organic ligand was successfully grafted on the surface of the magnetic silica nanoparticles. The fluorescent properties of the nanosensor were characterized and employed to detect Zn²⁺ with excellent selectivity and sensitivity (0.1 μM) toward Zn²⁺ over other cations even in trace amount.     

Folate conjugated fluorescent silica nanoparticles for labeling neoplastic cells

We describe a novel technique of using fluorescent silica nanoparticles (FSNPs) to detect over-expressed folate receptors, as typical for certain malignancies (metastatic adenocarcinoma, pituitary adenoma and others). Using St?ber's method with some modification, 135 nm size FSNPs were synthesized by a hydrolysis and co-condensation reaction of tetraethylorthosilicate (TEOS), fluorescein labeled (3-aminopropyl)triethoxysilane (APTS) and a water-dispersible silane reagent, (3-trihydroxysilyl)propyl methylphosphonate (THPMP) in the presence of ammonium hydroxide catalyst. Folic acid (folate) was covalently attached to the amine modified FSNPs by a carbodiimide coupling reaction. The characterization of folate-FSNPs was performed using a variety of spectroscopic (UV-VIS and fluorescence), microscopic (transmission electron microscopy, TEM) and light scattering techniques. Folate conjugated FSNPs were then targeted to human squamous cancer cells (SCC-9). Laser scanning confocal images successfully demonstrated the labeling of SCC-9 cells and the efficacy of FSNP based detection system.     

Bright and stable core-shell fluorescent silica nanoparticles

A class of highly fluorescent and photostable core-shell nanoparticles from a modified Stober synthesis in the size range of 20-30 nm is described. These nanoparticles are monodisperse in solution, 20 times brighter, and more photostable than their constituent fluorophore, and are amenable to specific labeling of biological macromolecules for bioimaging experiments. The photophysical characteristics of the encapsulated fluorophores differ from their solution properties. This raises the possibility of tuning nanoparticle structure toward enhanced radiative properties, making them an attractive material platform for a diverse range of applications.     

In situ synthesis and magnetic studies of iron oxide nanoparticles in calcium-alginate matrix for biomedical applications

In this work we applied a new route to synthesize magnetic iron oxide nanoparticles into alginate polymer for future application as drug delivery system activated by magnetic external stimuli. Calcium-alginate was used to encapsulate iron oxide nanoparticles, and as scaffold for particle nucleation and its influence on particles size and magnetic properties were studied. The iron oxide mean sizes were between 4.3 and 9.5 nm. Iron is dispersed throughout the polymer matrix mainly as iron oxide particles, and a small fraction as iron (III) occupying calcium sites in the polymer network. The temperature dependence of the Mössbauer spectra is typical of superparamagnetic particles in agreement with the magnetic susceptibility data.     

A novel fluorescent label based on organic dye-doped silica nanoparticles for HepG liver cancer cell recognition

In this paper, we report a method for the recognition of HepG liver cancer cells with the use of a novel fluorescent label based on organic dye-doped fluorescent silica nanoparticles. The novel organic dye-doped silica nanoparticles are prepared with a water-in-oil microemulsion technique. The silica network is produced by the controlled synchronous hydrolysis of tetraethoxysilane and 3-amino-propyltriethoxysilane (APTES). The organic dye fluorescein isothiocyanate is doped inside as a luminescent signaling element, through covalent bonding to the amino group of APTES. The organic dye-doped core-shell nanoparticles are highly luminescent and exhibit minimal dye leaching and excellent photostability. A novel fluorescent label method based on biological fluorescent nanoparticles has been developed. The dye-doped fluorescent silica nanoparticles are covalently immobilized with anti-human liver cancer monoclonal antibody HAb18. We have used antibody-labeled fluorescent nanoparticles to recognize HepG liver cancer cells. It has been observed that the bioassay based on the organic dye-doped nanoparticles can identify the target cells selectively and efficiently. The fluorescent nanoparticle label also exhibits high photostability.     

Mesoporous silica nanoparticles in nanomedicine applications

In the last few years mesoporous silica nanoparticles (MSNs) have gained the attention of the nanomedicine research community, especially for the potential treatment of cancer. Although this topic has been reviewed before, periodic updates on such a hot topic are necessary due to the dynamic character of this field. The reasons that make MSNs so attractive for designing controlled drug delivery systems lie beneath their physico-chemical stability, easy functionalisation, low toxicity and their great loading capacity of many different types of therapeutic agents. The present brief overview tries to cover some of the recent findings on stimuli-responsive mesoporous silica nanocarriers together with the efforts to design targeted nanosystems using that platform. The versatility of those smart nanocarriers has promoted them as very promising candidates to be used in the clinic in the near future to overcome some of the pitfalls of conventional medicine.

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Synthesis and sintering of biomimetic hydroxyapatite nanoparticles for biomedical applications

Synthesis of monodisperse nanoparticles with uniform morphology and narrow size distribution as achieved by nature is a challenge to materials scientists. Mimicking the process of biomineralization has led to the development of biomolecules mediated synthesis of nanoparticles that overcomes many of the problems associated with nanoparticle synthesis. Termed as biomimetics this paradigm shift in the philosophy of synthesis of materials is very advantageous for the design-based synthesis of nanoparticles. The effect of concentration of a protein named bovine serum albumin on particle size, morphology and degree of crystallinity of biomimetically synthesized hydroxyapatite particles, has been studied. Results establish 0.5% protein as the required concentration to produce 30–40 nm sized hydroxyapatite particles with an optimum degree of crystallinity as required for biomedical applications. These particles synthesized under certain stringent conditions are found to have stoichiometric calcium:phosphorus ratio of 1.67 and exhibit restricted grain growth during sintering.     

Synthesis and physical characterization of magnetite nanoparticles for biomedical applications

Iron oxide nanoparticles for biomedical applications in the size range of 15–130 nm were prepared by either oxidative hydrolysis of ferrous sulfate with KOH or precipitation from ferrous/ferric chloride solutions. The magnetite particle size is controlled by variation of pH and temperature. The synthesized magnetite nanoparticles are partially oxidized as signaled by ferrous concentrations of below 24 wt% Fe²⁺ and lattice parameters of a₀ ≤ 8.39 Å which are smaller compared to 8.39 Å for stoichiometric magnetite. The extend of oxidation increases with decreasing particle size. Heating at 150–350 °C topotactically transforms the magnetite nanoparticles into stoichiometric tetragonal maghemite (ferrous ion concentration c_Fe²⁺=0 and a₀ = 8.34 Å) without significant particle growth. The magnetite–maghemite transformation is studied with thermal analysis, XRD and IR spectroscopy. The saturation magnetizations of the magnetite and maghemite particles decrease with decreasing particle size. The variation of M_s with particle size is interpreted using a magnetic core–shell particle model. Magnetite particles with d ≤ 16 nm show superparamagnetic behavior at room temperature whereas particles with diameter >16 nm display hysteresis behavior. These particles are candidates for biomedical applications, e.g. controlled drug release or hyperthermia.     

Imaging gold nanoparticles for DNA sequence recognition in biomedical applications

The hybridization of single-stranded oligonucleotide-derivatized gold nanoparticles (Au nanoprobes) with double stranded complementary DNA was directly observed by atomic force microscopy (AFM). This specific interaction is the basis for an Au nanoprobe-based homogeneous assay for specific DNA sequence detection, based on salt-induced particle aggregation that is prevented when a complementary target is present. For long DNA targets (linearized plasmid DNA) complicated hybridized target DNA-Au-nanoprobes structures were formed, that were interpreted as the basis for stability of the Au nanoprobes against salt-induced aggregation. For shorter DNA targets (PCR amplified fragments) hybridization with the Au nanoprobes occurred, in the majority of cases, in the expected location of the DNA target fragment containing the specific sequence. The formation of the observed DNA hybridized structures provides evidence at the molecular level for specific hybridization to the target sequence as the method of binding of the Au nanoprobes.     

Magnetic and fluorescent core-shell nanoparticles for ratiometric pH sensing

This paper describes the preparation of nanoparticles composed of a magnetic core surrounded by two successive silica shells embedding two fluorophores, showing uniform nanoparticle size (50-60 nm in diameter) and shape, which allow ratiometric pH measurements in the pH range 5-8. Uncoated iron oxide magnetic nanoparticles (～10 nm in diameter) were formed by the coprecipitation reaction of ferrous and ferric salts. Then, they were added to a water-in-oil microemulsion where the hydrophilic silica shells were obtained through hydrolysis and condensation of tetraethoxyorthosilicate together with the corresponding silylated dye derivatives-a sulforhodamine was embedded in the inner silica shell and used as the reference dye while a pH-sensitive fluorescein was incorporated in the outer shell as the pH indicator. The magnetic nanoparticles were characterized using vibrating sample magnetometry, dynamic light scattering, transmission electron microscopy, x-ray diffraction and Fourier transform infrared spectroscopy. The relationship between the analytical parameter, that is, the ratio of fluorescence between the sensing and reference dyes versus the pH was adjusted to a sigmoidal fit using a Boltzmann type equation giving an apparent pK(a) value of 6.8. The fluorescence intensity of the reference dye did not change significantly (～3.0%) on modifying the pH of the nanoparticle dispersion. Finally, the proposed method was statistically validated against a reference procedure using samples of water and physiological buffer with 2% of horse serum, indicating that there are no significant statistical differences at a 95% confidence level.     

Synthesis and characterization of photoswitchable fluorescent silica nanoparticles

We have designed and synthesized a new functional (amino reactive) highly efficient fluorescent molecular switch (FMS) with a photochromic diarylethene and a rhodamine fluorescent dye. The reactive group in this FMS -N-hydroxysuccinimide ester- allows selective labeling of amino containing molecules or other materials. In ethanolic solutions, the compound displays a large fluorescent quantum yield of 52 % and a large fluorescence modulation ratio (94 %) between two states that may be interconverted with red and near-UV light. Silica nanoparticles incorporating the new FMS were prepared and characterized, and their spectroscopic and switching properties were also studied. The dye retained its properties after the incorporation into the silica, thereby allowing light-induced reversible high modulation of the fluorescence signal of a single particle for up to 60 cycles, before undergoing irreversible photobleaching. Some applications of these particles in fluorescence microscopy are also demonstrated. In particular, subdiffraction images of nanoparticles were obtained, in the focal plane of a confocal microscope.     

Fabrication of novel TiZr alloy foams for biomedical applications

Bone injuries and failures often require the inception of implant biomaterial. Research in this area has received increasing attention recently. In particular, porous metals are attractive due to its unique physical, mechanical, and new bone tissue ingrowth properties. In the present study, TiZr alloy powders were prepared using mechanical alloying. Novel TiZr alloy foams with relative densities of approximately 0.3 were fabricated by a powder metallurgical process. The TiZr alloy foams displayed an interconnected porous structure resembling bone and the pore size ranged from 200 to 500 μm. The compressive plateau stress and the Young’s modulus of the TiZr foam were 78.4 MPa and 15.3 GPa, respectively. Both the porous structure and the mechanical properties of the TiZr foam were very close to those of natural bone.