Long-range electrothermal fluid motion in microfluidic systems.

AC electrothermal flow (ACEF) is the fluid motion created as a result of Joule heating induced temperature gradients. ACEF is capable of performing major microfluidic operations, such as pumping, mixing, concentration, separation and assay enhancement, and is effective in biological samples with a wide range of electrical conductivity. Here, we report long-range fluid motion induced by ACEF, which creates centimeter-scale vortices. The long-range fluid motion displays a strong voltage dependence and is suppressed in microchannels with a characteristic length below ~300 µm. An extended computational model of ACEF, which considers the effects of the density gradient and temperature-dependent parameters, is developed and compared experimentally by particle image velocimetry. The model captures the essence of ACEF in a wide range of channel dimensions and operating conditions. The combined experimental and computational study reveals the essential roles of buoyancy, temperature rise, and associated changes in material properties in the formation of the long-range fluid motion. Our results provide critical information for the design and modeling of ACEF based microfluidic systems toward various bioanalytical applications.

Cellular self-organization by autocatalytic alignment feedback.

Myoblasts aggregate, differentiate and fuse to form skeletal muscle during both embryogenesis and tissue regeneration. For proper muscle function, long-range self-organization of myoblasts is required to create organized muscle architecture globally aligned to neighboring tissue. However, how the cells process geometric information over distances considerably longer than individual cells to self-organize into well-ordered, aligned and multinucleated myofibers remains a central question in developmental biology and regenerative medicine. Using plasma lithography micropatterning to create spatial cues for cell guidance, we show a physical mechanism by which orientation information can propagate for a long distance from a geometric boundary to guide development of muscle tissue. This long-range alignment occurs only in differentiating myoblasts, but not in non-fusing myoblasts perturbed by microfluidic disturbances or other non-fusing cell types. Computational cellular automata analysis of the spatiotemporal evolution of the self-organization process reveals that myogenic fusion in conjunction with rotational inertia functions in a self-reinforcing manner to enhance long-range propagation of alignment information. With this autocatalytic alignment feedback, well-ordered alignment of muscle could reinforce existing orientations and help promote proper arrangement with neighboring tissue and overall organization. Such physical self-enhancement might represent a fundamental mechanism for long-range pattern formation during tissue morphogenesis.

Dielectrophoresis-Mediated Electrodeformation as a Means of Determining Individual Platelet Stiffness.

Platelets, essential for hemostasis, are easily activated via biochemical and mechanical stimuli. Cell stiffness is a vital parameter modulating the mechano-transduction of exogenous mechanical stimuli. While methods exist to measure cell stiffness, no ready method exists for measuring platelet stiffness that is both minimally-contacting, imparting minimal exogenous force and non-activating. We developed a minimal-contact methodology capable of trapping and measuring the stiffness of individual platelets utilizing dielectrophoresis (DEP)-mediated electrodeformation. Parametric studies demonstrate a non-uniform electric field in the MHz frequency range (0.2-20 MHz) is required for generating effective DEP forces on platelets, suspended in isotonic buffer with conductivity ~100-200 µS/cm. A nano-Newton DEP force (0.125-4.5 nN) was demonstrated to be essential for platelet electrodeformation, which could be generated with an electric field with strength of 1.5-9 V/µm. Young's moduli of platelets were calculated using a Maxwell stress tensor model and stress-deformation relationship. Platelet stiffness was determined to be in the range of 3.5 ± 1.4 and 8.5 ± 1.5 kPa for resting and 0.4% paraformaldehyde-treated cells, respectively. The developed methodology fills a gap in approaches of measuring individual platelet stiffness, free of inadvertent platelet activation, which will facilitate further studies of mechanisms involved in mechanically-mediated platelet activation.

Comparative study of antibody immobilization mediated by lipid and polymer fibers.

Antibody immobilization and function retention are important to a variety of applications, including proteomics, drug discovery, diagnostics, and biosensors. The present study investigates antibody immobilization mediated by cholesteryl succinyl silane (CSS) fibers, in comparison to hydrophobic polycaprolactone (PCL) fibers and hydrophilic plasma-treated PCL fibers. When incubated with a model protein, the formation of protein aggregates is observed on hydrophobic PCL fibers but not on the more hydrophobic CSS fibers, indicating that CSS fibers immobilize proteins through mechanisms other than hydrophobic interaction. When exposed to a limited amount of antibody, CSS fibers immobilize more antibodies than plasma-treated PCL fibers and no fewer antibodies than PCL fibers. The function retention of antibodies immobilized on the fibers is analyzed using a cell-capture assay, which shows that the antibody-functionalized CSS fibrous matrices capture 6- or 7-fold more cells than the antibody-functionalized PCL or plasma-treated PCL fibrous matrices, respectively. Data collected from the study show that the lipid fiber-mediated immobilization of antibody not only maintains the advantages of physical immobilization such as easiness and rapidness of operation but also improves function retention.

Anti-EGFR antibody conjugated organic-inorganic hybrid lipid nanovesicles selectively target tumor cells.

Chemical conjugation of anti-epidermal growth factor receptor monoclonal antibodies (anti-EGFR mAbs) to organic-inorganic hybrid liposomal immunocerasomes via maleimide-thiol coupling chemistry is explored as a mechanism for selectively targeting cancer cells. The cellular uptake and internalization of immunocerasomes are investigated in A431 cells that express an abnormally high level of EGFR, DU145 cells that overexpress EGFR, and HL-60 cells that are used as a negative control. The internalization study reveals a strong correlation between the receptor-mediated endocytosis of immunocerasomes and the membrane expression of EGFR. Further, free anti-EGFR mAbs and immunocerasomes conjugated with anti-EGFR mAbs at nanomolar doses display similar anti-proliferative effects on A431 cells. Additionally, serum proteins greatly reduce the cellular uptake of cerasomes that is mediated by non-specific receptors, but have no adverse effects on the specific EGFR-mediated delivery of immunocerasomes to A431 cells.

Medication adherence: is it a hidden drug-related problem in hidden elderly?

AIM: The present study aimed to identify the health needs of the hidden elderly in Hong Kong, and to investigate the impacts of pharmacist and nursing interventions on medication management and well-being in hidden elderly. METHODS: Participants were recruited by social workers if they were aged 65 years or older; did not have normal social life and network; did not have family support; and were not linked to the existing network of community support. Pharmacists identified drug-related problems. The health needs of participants were assessed by observations and interviews. Outcome measurements were scores of Morisky 8-item Medication Adherence Scale and EuroQoL (Quality of Life) 5-D Questionnaire. RESULTS: A total of 93 participants were recruited and 86 participants completed the study. The mean age was 81.46 ± 5.70 years, the mean number of chronic disease was 3.29 ± 1.45 and the mean number of chronic medications was 6.36 ± 2.96. The most commonly observed chronic diseases were hypertension, cardiac problems, diabetes, hyperlipidemia and arthritis. Drug non-adherence and storage problems were found in 61.63% and 69.77% of participants. The mean total EuroQoL score increased by 1.05 (P ≤ 0.001). The mean Morisky score decreased by 0.61, signifying improvement of medication adherence (P< 0.001). Female sex (P = 0.045), polypharmacy with more than nine concurrent medications (P = 0.013), arthritis (P = 0.006) and drug storage problems (P = 0.002) were identified as factors associated with poor medication adherence. CONCLUSIONS: The majority of hidden elderly suffered from multiple chronic diseases, and the prevalence of drug-related problems was high. Pharmacist and nursing interventions improved drug-related problems, drug compliance and quality of life in hidden elderly.

Centering of organic-inorganic hybrid liposomal cerasomes in electrospun gelatin nanofibers.

A study investigating the embedding of stabilized organic-inorganic liposomal cerasomes in gelatin nanofibers through the electrospinning of cerasome-dispersed gelatin aqueous solution is presented. Fluorescent and transmission electron microscopy confirm the embedding and centering of cerasomes in the electrospun nanofibers. A simple mechanism is proposed for the centering of cerasomes in gelatin nanofibers. The ability to incorporate cerasomes capable of encapsulating a variety of bioactive molecules provides a promising method to functionalize polymer nanofibers.

Plasma lithography surface patterning for creation of cell networks.

Systematic manipulation of a cell microenvironment with micro- and nanoscale resolution is often required for deciphering various cellular and molecular phenomena. To address this requirement, we have developed a plasma lithography technique to manipulate the cellular microenvironment by creating a patterned surface with feature sizes ranging from 100 nm to millimeters. The goal of this technique is to be able to study, in a controlled way, the behaviors of individual cells as well as groups of cells and their interactions. This plasma lithography method is based on selective modification of the surface chemistry on a substrate by means of shielding the contact of low-temperature plasma with a physical mold. This selective shielding leaves a chemical pattern which can guide cell attachment and movement. This pattern, or surface template, can then be used to create networks of cells whose structure can mimic that found in nature and produces a controllable environment for experimental investigations. The technique is well suited to studying biological phenomenon as it produces stable surface patterns on transparent polymeric substrates in a biocompatible manner. The surface patterns last for weeks to months and can thus guide interaction with cells for long time periods which facilitates the study of long-term cellular processes, such as differentiation and adaption. The modification to the surface is primarily chemical in nature and thus does not introduce topographical or physical interference for interpretation of results. It also does not involve any harsh or toxic substances to achieve patterning and is compatible for tissue culture. Furthermore, it can be applied to modify various types of polymeric substrates, which due to the ability to tune their properties are ideal for and are widely used in biological applications. The resolution achievable is also beneficial, as isolation of specific processes such as migration, adhesion, or binding allows for discrete, clear observations at the single to multicell level. This method has been employed to form diverse networks of different cell types for investigations involving migration, signaling, tissue formation, and the behavior and interactions of neurons arraigned in a network.