Comparison of cultured cell attachment on a temperature-responsive polymer, poly-L-lysine, and collagen using modeling curves and a thermal-controlled quartz crystal microbalance.

The characteristics of cultured cell attachment onto poly-L-lysine (PLL), collagen, and the thermoresponsive polymer poly(N-isopropylacrylamide) (PNIPAM) were studied using a quartz crystal microbalance (QCM). A QCM with microscope cameras enclosed in a Peltier chamber was developed to enable QCM measurements and microphotographic imaging to be conducted in a temperature-controlled CO2 incubator. Human hepatoma cell line HepG2 cells were cultured on the quartz crystals coated with PLL, collagen, and PNIPAM. Response curves of the resonant frequency of the quartz crystals during the cell attachment process were analyzed on the basis of the parameters of modeling curves fit to the experimentally obtained curves. Analysis of the fitting curves showed that the time constants of the first-lag response were 11 h for PLL, 16 h for collagen, and 38 h for PNIPAM and that the frequency change for the PNIPAM films was six times smaller than those for the PLL and collagen films. These findings were supported by photographic images showing wider cell spread on PLL and collagen than on PNIPAM. The response of cells on PNIPAM was measured during a thermal cycle from 37 to 20 °C to 37 °C. In the resonance frequency-resonance resistance (F-R) diagram, the slopes of ΔR/ΔF corresponding to the cell attachment process and those corresponding to the thermal cycling process differed; the positions in the F-R diagram also shifted to higher resonant frequencies after the thermal cycle. These results suggested that the mass effect decreased as a result of the weakening of the cell attachment strength by the thermal cycle because the molecular brushes of PNIPAM were disarranged.

Triboelectric charging of polytetrafluoroethylene antithrombotic catheters.

This study proposes that a polytetrafluoroethylene (PTFE) electret tube charged by frictional electricity can prevent the solidification of the indwelling catheter in blood vessels. Coagulation in intravascular indwelling catheters may discontinue the treatment because of thrombus-derived bacteria-adhesion infections or poor blood removal. Current commercially available intravascular catheters lack complete antithrombotic measures, even with heparin or urokinase antithrombotic coatings. Herein, we tested the effectiveness of an antithrombotic treatment that prevents coagulation using a static electric charge on the interior of the PTFE tube via the triboelectric effect by rubbing the tube's inner wall with a round glass rod. The anticoagulation properties were evaluated by enclosing a sample of blood in an electret tube and observing the coagulase adhering to the inner wall using a microscope. To confirm the effectiveness of this treatment, the charge-distribution on the inner surface of the electret tube was measured, surface irregularities were observed, and the elements on the surface were analyzed. The surface potential inside the electret tube was - 366.4 V, which proved effective for an antithrombotic treatment, as it discouraged coagulation, and the triboelectric charging process caused neither surface element denaturation nor significant surface irregularities. The nearly uniform negative surface charge on the inside of the tube was responsible for the antithrombotic effect because no surface irregularities or change in the surface element denaturation was observed. Triboelectrically charged PTFE electret tubes are highly useful for intravascular indwelling catheters.

Effect of arching spine on deformation of the ligamentum flavum during epidural needle insertion.

When administering epidural anesthesia, anesthesiologists ask patients to arch their back. Arching the spine is thought to enlarge the gap between neighboring vertebral bones. The author hypothesized that tension inside the ligamentum flavum generated by arching the spine would reduce deformation of the ligamentum flavum during epidural needle insertion. Porcine spines from a slaughterhouse were cut at the vertebral body and separated into 10 pieces. Ligamentum flavum was painted black with liquid ink. A CCD camera recorded the deformation of the ligamentum flavum during needle insertion. To simulate the arching spine, the width of the ligamentum flavum was enlarged by a retractor. The thickness of the ligamentum flavum was measured from the stained images by the Elastica-van Gieson method. The maximum reaction force showed no significant difference between the natural and the enlarged ligamentum flavum. Average deformation was observed a statically significant decrease between the naturally deformed and retractor-enlarged ligamentum flavum. For the maximum reaction force the coefficient of variance decreased by dividing raw data with thickness of the ligamentum flavum, which meant that the maximum reaction force correlated with thickness of the ligamentum flavum. Less effect on deformation was observed. Hypothesis was correct in the porcine study, while the difference between the porcine and the patient's spine should be examined in the next research.

An enzyme-chromogenic surface plasmon resonance biosensor probe for hydrogen peroxide determination using a modified Trinder's reagent.

An absorption-based surface plasmon resonance (SPR(Abs)) biosensor probe has been developed for simple and reproducible measurements of hydrogen peroxide using a modified Trinder's reagent (a chromogenic reagent). The reagent enabled the determination of the hydrogen peroxide concentration by the development of deep color dyes (lambda(max)=630 nm) through the oxidative coupling reaction with N-ethyl-N-(2-hydroxy-3-sulfopropyl)-3,5-dimethylaniline sodium salt monohydrate (MAOS; C(13)H(20)NNaO(4)S.H(2)O) and 4-aminoantipyrine (4-AA) in the presence of hydrogen peroxide and horseradish peroxidase (HRP). In the present study, urea as an adduct of hydrogen peroxide for color development could be omitted from the measurement solution. The measurement solution containing 5mM hydrogen peroxide was deeply colored at a high absorbance value calculated as 46.7cm(-1) and was directly applied to the SPR(Abs) biosensing without dilution. The measurement was simply performed by dropping the measurement solution onto the surface of the SPR sensor probe, and the SPR(Abs) biosensor response to hydrogen peroxide was obtained as a reflectivity change in the SPR spectrum. After investigation of the pH profiles in the SPR(Abs) biosensor probe, a linear calibration curve was obtained between 1.0 and 50mM hydrogen peroxide (r=0.991, six points, average of relative standard deviation; 0.152%, n=3) with a detection limit of 0.5mM. To examine the applicability of this SPR(Abs) biosensor probe, 20mM glucose detection using glucose oxidase was also confirmed without influence of the refractive index in the measurement solution. Thus, the SPR(Abs) biosensor probe employing the modified Trinder's reagent demonstrated applicability to other analyte biosensing tools.