Proliferation of SH-SY5Y neuroblastoma cells on confined spaces.

BACKGROUND: Microfluidics offers precise drug delivery and continuous monitoring of cell functions, which is crucial for studying the effects of toxins and drugs. Ensuring proper cell growth in these space-constrained systems is essential for obtaining consistent results comparable to standard Petri dishes. NEW METHOD: We investigated the proliferation of SH-SY5Y cells on circular polycarbonate chambers with varying surface areas. SH-SY5Y cells were chosen for their relevance in neurodegenerative disease research. RESULTS: Our study demonstrates a correlation between the chamber surface area and SH-SY5Y cell growth rates. Cells cultured in chambers larger than 10 mm in diameter exhibited growth comparable to standard 60-mm dishes. In contrast, smaller chambers significantly impeded growth, even at identical seeding densities. Similar patterns were observed for HeLaGFP cells, while 16HBE14σ cells proliferated efficiently regardless of chamber size. Additionally, SH-SY5Y cells were studied in a 12-mm diameter sealed chamber to assess growth under restricted gas exchange conditions. COMPARISON WITH EXISTING METHODS: Our findings underscore the limitations of small chamber sizes in microfluidic systems for SH-SY5Y cells, an issue not typically addressed by conventional methods. CONCLUSIONS: SH-SY5Y cell growth is highly sensitive to spatial constraints, with markedly reduced proliferation in chambers smaller than 10 mm. This highlights the need to carefully consider chamber size in microfluidic experiments to achieve cell growth rates comparable to standard culture dishes. The study also shows that while SH-SY5Y and HeLaGFP cells are affected by chamber size, 16HBE14σ cells are not. These insights are vital for designing effective microfluidic systems for bioengineering research.

Mitochondrial Dynamics in Neurodegenerative Diseases: Unraveling the Role of Fusion and Fission Processes.

Neurodegenerative diseases (NDs) are a diverse group of disorders characterized by the progressive degeneration and death of neurons, leading to a range of neurological symptoms. Despite the heterogeneity of these conditions, a common denominator is the implication of mitochondrial dysfunction in their pathogenesis. Mitochondria play a crucial role in creating biomolecules, providing energy through adenosine triphosphate (ATP) generated by oxidative phosphorylation (OXPHOS), and producing reactive oxygen species (ROS). When they're not functioning correctly, becoming fragmented and losing their membrane potential, they contribute to these diseases. In this review, we explore how mitochondria fuse and undergo fission, especially in the context of NDs. We discuss the genetic and protein mutations linked to these diseases and how they impact mitochondrial dynamics. We also look at the key regulatory proteins in fusion (MFN1, MFN2, and OPA1) and fission (DRP1 and FIS1), including their post-translational modifications. Furthermore, we highlight potential drugs that can influence mitochondrial dynamics. By unpacking these complex processes, we aim to direct research towards treatments that can improve life quality for people with these challenging conditions.

Effective Optical Image Assessment of Cellulose Paper Immunostrips for Blood Typing.

Novel high-performance biosensing devices, based on a microporous cellulose matrix, have been of great interest due to their high sensitivity, low cost, and simple operation. Herein, we report on the design and testing of portable paper-based immunostrips (IMS) for in-field blood typing in emergencies requiring blood transfusion. Cellulose fibrils of a paper membrane were functionalized with antibodies via supramolecular interactions. The formation of hydrogen bonds between IgM pentamer and cellulose fibers was corroborated using quantum mechanical calculations with a model cellulose chain and a representative amino acid sequence. In the proposed immunostrips, paper with a pore size of 3 µm dia. was used to enable functionalization of its channels with antibody molecules while blocking the red blood cells (RBC) from channel entering. Under the optimized test conditions, all blood types of AB0 and Rh system could be determined by naked eye examination, requiring only a small blood sample (3.5 µL). The durability of IgM immunostrips against storing has been tested. A new method of statistical evaluation of digitized blood agglutination images, compatible with a clinical five-level system, has been proposed. Critical parameters of the agglutination process have been established to enable future development of automatic blood typing with machine vision and digital data processing.

A Seedless Method for Gold Nanoparticle Growth inside a Silica Matrix: Fabrication of Materials Capable of Third-Harmonic Generation in the Near-Infrared.

A composite in which gold nanoparticles (AuNPs) approximately 10 nm in size are embedded in amorphous transparent silica matrix has been produced. The synthetic protocol uses HAuCl4 as the Au ion source, tetraethoxysilane (TEOS) as the SiO2 precursor, and l-ascorbic acid (AA) as the reducing agent. AA is employed before the sol-gel process in an amount sufficient only for reduction of Au3+ ions to Au+ . By using a cationic surfactant, benzylcetyldimethylammonium chloride hydrate (BDAC) and/or cetyltrimethylammonium bromide (CTAB), the Au+ ions are encapsulated within metalomicelles, which prevents them from being reduced to Au0 and enables their homogeneous distribution in the gel. Reduction of Au+ to Au0 and the growth of the AuNPs occurs at room temperature during the gelation, and arises from the release of EtOH during the hydrolysis of TEOS. The composites contain 0.027 wt % of Au. They exhibit nonlinear optical behavior characterized by the third-order nonlinear refraction index, n2 , in the range 3.6-5.7×10-16  cm2 W-1 at λ=1.030 µm. The composites are capable of effective third-harmonic generation of ultrashort near-IR (210 fs, 1.030 µm) laser pulse through a direct third-order mechanism.

Length-scale dependent transport properties of colloidal and protein solutions for prediction of crystal nucleation rates.

We propose a scaling equation describing transport properties (diffusion and viscosity) in the solutions of colloidal particles. We apply the equation to 23 different systems including colloids and proteins differing in size (range of diameters: 4 nm to 1 µm), and volume fractions (10(-3)-0.56). In solutions under study colloids/proteins interact via steric, hydrodynamic, van der Waals and/or electrostatic interactions. We implement contribution of those interactions into the scaling law. Finally we use our scaling law together with the literature values of the barrier for nucleation to predict crystal nucleation rates of hard-sphere like colloids. The resulting crystal nucleation rates agree with existing experimental data.

A "wrap-and-wrest" mechanism of fluorescence quenching of CdSe/ZnS quantum dots by surfactant molecules.

We identified a mechanism of fluorescence quenching of CdSe/ZnS quantum dots (QDs) coated with two organic layers, octadecylamine and an amphiphilic polymer containing COOH groups, by nonionic polyoxyethylene-based (C12En) surfactants. The surfactant molecules by themselves do not affect the fluorescence of the QDs. For the quenching to occur, "wrapping" of the QDs by a bilayer of the surfactant molecules is necessary. The formation of the bilayer causes an irreversible detachment ("wresting") of the ligand molecules, accompanied by the creation of quenching sites on the QD surface. Due to its two-stage nature, we refer to the quenching mechanism as the "wrap-and-wrest" mechanism. The adsorption of the surfactant on the QD surface is a relatively slow process, occurring within minutes or hours. Such long quenching times allowed monitoring surfactant adsorption progress in real time. The fluorescence signal decays exponentially, and the decay time is inversely proportional to the surfactant concentration in solution.

New one-pot technique to introduce charged nanoparticles into a lyotropic liquid crystal matrix.

We present a new method to incorporate hydrophilic charged nanoparticles into the lyotropic liquid crystal (LLC) template. This method is based on the effect of the polymer-induced phase separation (PIPS) and consists of two steps. In the first step, the nanoparticles are mixed with a surfactant micellar solution. In the second step, upon addition of polymer, phase separation is induced and the LLC phase doped with the nanoparticles is formed. Columnar hexagonal and lamellar LLC templates are obtained with the PIPS method. The ordering of the LLC phase can be controlled by the amount of polymer added to induce phase separation. The method works both for the system of nonionic surfactants and polymers and ionic surfactants and polyelectrolytes. We demonstrate that the PIPS method enables the fabrication of the LLC templates doped with positively or negatively charged nanoparticles as well as with a mixture of oppositely charged nanoparticles in arbitrary proportions.

Reverse vesicles from a salt-free catanionic surfactant system: a confocal fluorescence microscopy study.

We give a detailed confocal fluorescence microscopy study on reverse vesicles from a salt-free catanionic surfactant system. When tetradecyltrimethylammonium laurate (TTAL) and lauric acid (LA) are mixed in cyclohexane at the presence of a small amount of water, stable reverse vesicular phases form spontaneously. The reverse vesicular phases can be easily labeled with dyes of varying molecular size and hydrophobicity while the dyes are nearly insoluble in cyclohexane without reverse vesicles. This indicates the reverse vesicular phases can be good candidates to host guest molecules. With the help of a fluorescence microscope combined a confocal method, the features of these interesting reverse supramolecular self-assemblies were revealed for the first time. Because of the absence of electrostatic repulsions and hydration forces between adjacent vesicles, the reverse vesicles have a strong propensity to aggregate with each other and form three-dimensional clusters. The size distributions of both individual reverse vesicles and clusters are polydisperse. Huge multilamellar reverse vesicles with closely stacked thick walls (giant reverse onions) were observed. Besides the spherical reverse vesicles and onions, other supramolecular structures such as tubes have also been detected and structural evolutions between different structures were noticed. These interesting supramolecular self-assemblies form in a nonpolar organic solvent may serve as ideal micro- or nanoreaction centers for biological reactions and synthesis of inorganic nanomaterials.

Single-walled carbon nanotube/lyotropic liquid crystal hybrid materials fabricated by a phase separation method in the presence of polyelectrolyte.

We present a detailed study on the incorporation of single-walled carbon nanotubes (SWNTs) into lyotropic liquid crystals (LLC) by phase separation in the presence of polyelectrolytes. Two cases were studied in this work: (i) incorporation of SWNTs into the LLC phase formed by an anionic surfactant sodium dodecyl sulfate (SDS) in the presence of an anionic polyelectrolyte poly(sodium styrenesulfonate) (PSS); (ii) incorporation of SWNTs into the LLC phase formed by a cationic surfactant cetyltrimethylammonium bromide (CTAB) in the presence of a cationic polyelectrolyte poly(diallydimethylammonium chloride) (PDADMAC). The SWNTs/LLC composites were characterized by polarized optical microscopy (POM) observations and small-angle X-ray scattering (SAXS) measurements. In both systems, the surfactant phase was condensed into a hexagonal lattice by the polyelectrolyte within the investigated concentration range. Several factors that can influence the property of SWNTs/LLC composite were examined, including concentration of surfactants and polyelectrolytes and temperature. Aggregated SWNTs were not observed, indicating that SWNTs were well dispersed in the LLC phases. SAXS measurements showed the lattice parameter of the host LLC phase changed upon varying the mixing ratio of polyelectrolyte to ionic surfactant. The SWNTs/LLC hybrids showed considerable stability against temperature rise in both systems, and desorption of surfactant from SWNTs was not observed at higher temperature.

Incorporation of carbon nanotubes into a lyotropic liquid crystal by phase separation in the presence of a hydrophilic polymer.

Single-walled carbon nanotubes (SWNTs) were incorporated into a lyotropic liquid crystal (LLC) matrix formed by n-dodecyl octaoxyethene monoether (C(12)E(6)) at room temperature through spontaneous phase separation induced by nonionic hydrophilic polymer poly(ethylene glycol) (PEG). The quality of SWNTs/LLC composite was evaluated by polarized microscopy observations and small-angle X-ray scattering (SAXS) measurements. The results obtained clearly indicated that SWNTs have been successfully incorporated into the LLC matrix up to a considerable high content without destroying the LLC matrix, although interesting changes of the LLC matrix were also induced by SWNTs incorporation. By varying the ratio of PEG to C(12)E(6), the type of LLC matrix can be controlled from hexagonal phase to lamellar phase. Temperature was found to have a significant influence on the quality of SWNTs/LLC composite, and tube aggregation can be induced at higher temperature. When SWNTs were changed to multiwalled carbon nanotubes (MWNTs), they became difficult to be incorporated into LLC matrix because of an increase in the average tube diameter.

Polymer-induced ordering and phase separation in ionic surfactants.

We present a new method to induce phase separation in solutions of ionic surfactants. In this method, the phase separation is obtained either by addition of polyelectrolytes or nonionic polymers along with inorganic salt. As a result, the system separates into polyelectrolyte-rich (or nonionic polymer-rich) and surfactant-rich phase. Four types of the mixtures were investigated: (i) anionic surfactants and anionic polyelectrolytes, (ii) cationic surfactants and cationic polyelectrolytes, (iii) cationic surfactants and nonionic polymers, and (iv) anionic surfactants and nonionic polymers. We found that the addition of polyelectrolyte with the charge of the same sign as that of surfactant can induce the phase separation in a wide range of surfactant concentrations. The addition of nonionic polymers induces the phase separation only in solutions of cationic surfactants. Moreover, the addition of nonionic polymers induces the phase separation only for relatively high total content of polymer and surfactant in the mixture. We found however that the addition of inorganic salt to the mixture of cationic surfactant and nonionic polymer triggers the phase separation even for a small concentrations of surfactant. In our experiments, water as well as mixtures of water and polar solvents were employed as solvents. Based on the optical microscopy studies we found that the surfactant-rich phase represents hexagonal ordering.

Scaling form of viscosity at all length-scales in poly(ethylene glycol) solutions studied by fluorescence correlation spectroscopy and capillary electrophoresis.

We measured the viscosity of poly(ethylene glycol) (PEG 6000, 12,000, 20,000) in water using capillary electrophoresis and fluorescence correlation spectroscopy with nanoscopic probes of different diameters (from 1.7 to 114 nm). For a probe of diameter smaller than the radius of gyration of PEG (e.g. rhodamine B or lyzozyme) the measured nanoviscosity was orders of magnitude smaller than the macroviscosity. For sizes equal to (or larger than) the polymer radius of gyration, macroscopic value of viscosity was measured. A mathematical relation for macro and nanoviscosity was found as a function of PEG radius of gyration, R(g), correlation length in semi-dilute solution, xi, and probe size, R. For R < R(g), the nanoviscosity (normalized by water viscosity) is given by exp(b(R/xi)a), and for R > R(g), both nano and macroviscosity follow the same curve, exp(b(R/xi)a), where a and b are two constants close to unity. This mathematical relation was shown to equally well describe rhodamine (of size 1.7 nm) in PEG 20,000 and the macroviscosity of PEG 8,000,000, whose radius of gyration exceeds 200 nm. Additionally, for the smallest probes (rhodamine B and lysozyme) we have verified, using capillary electrophoresis and fluorescence correlation spectroscopy, that the Stokes-Einstein (SE) relation holds, providing that we use a size-dependent viscosity in the formula. The SE relation is correct even in PEG solutions of very high viscosity (three orders of magnitude larger than that of water).