Self-assembled monolayers on polymer surfaces

The method of forming self-assembled monolayers on polymer surfaces is reviewed. It is shown that the alkylsiloxane monolayers formed on polymer surfaces are structurally similar to the analogous monolayers formed on silicon wafers. These monolayers can be post-derivatized in order to introduce a number of hydrophobic, reactive and polar functionalities, some of which may be useful model systems for studying biological interactions at surfaces.     

Statistical method for surface pattern-matching between dissimilar molecules: electrostatic potentials and accessible surfaces

This paper outlines a statistical method for patternmatching between surfaces and is applicable to structural and energetic patterns found on molecular surfaces. Correlation coefficients generated for the pattern match are scale invariant. Regression analysis applied to the patterns reveals the scaling and displacement relationships. The method for measuring the similarities between molecular surfaces of two dissimilar molecules held infixed orientations is given explicitly. Implicit in this procedure is a method for studying the inverse phenomenon, namely complementarity between surface parameters at a binding site and its ligand. The method has been used to assess surface differences in structural similarities generated by computer fitting and by visual comparison. Various pitfalls likely to be encountered in evaluating molecular structural similarities are noted.     

An Improved Mechanical Testing Method to Assess Bone-implant Anchorage

Recent advances in material science have led to a substantial increase in the topographical complexity of implant surfaces, both on a micro- and a nano-scale. As such, traditional methods of describing implant surfaces - namely numerical determinants of surface roughness - are inadequate for predicting in vivo performance. Biomechanical testing provides an accurate and comparative platform to analyze the performance of biomaterial surfaces. An improved mechanical testing method to test the anchorage of bone to candidate implant surfaces is presented. The method is applicable to both early and later stages of healing and can be employed for any range of chemically or mechanically modified surfaces - but not smooth surfaces. Custom rectangular implants are placed bilaterally in the distal femora of male Wistar rats and collected with the surrounding bone. Test specimens are prepared and potted using a novel breakaway mold and the disruption test is conducted using a mechanical testing machine. This method allows for alignment of the disruption force exactly perpendicular, or parallel, to the plane of the implant surface, and provides an accurate and reproducible means for isolating an exact peri-implant region for testing.     

Estimation of amino acid residue substitution rates at local spatial regions and application in protein function inference: a Bayesian Monte Carlo approach

The amino acid sequences of proteins provide rich information for inferring distant phylogenetic relationships and for predicting protein functions. Estimating the rate matrix of residue substitutions from amino acid sequences is also important because the rate matrix can be used to develop scoring matrices for sequence alignment. Here we use a continuous time Markov process to model the substitution rates of residues and develop a Bayesian Markov chain Monte Carlo method for rate estimation. We validate our method using simulated artificial protein sequences. Because different local regions such as binding surfaces and the protein interior core experience different selection pressures due to functional or stability constraints, we use our method to estimate the substitution rates of local regions. Our results show that the substitution rates are very different for residues in the buried core and residues on the solvent-exposed surfaces. In addition, the rest of the proteins on the binding surfaces also have very different substitution rates from residues. Based on these findings, we further develop a method for protein function prediction by surface matching using scoring matrices derived from estimated substitution rates for residues located on the binding surfaces. We show with examples that our method is effective in identifying functionally related proteins that have overall low sequence identity, a task known to be very challenging.     

Protein-protein recognition: method for finding complementary surfaces of interacting proteins

A new method is described for searching for complementary surfaces of protein molecules from known coordinates of their non-hydrogen atoms. Each atom is assigned an arbitrary feature which is specific of its interactions with other atoms. The plots representing surfaces are generated. For each pair of surfaces a number of coincidences is calculated which increases as the number of contacts is increased between these atom pairs whose contribution to the energy of interaction is considered to be essential. The results obtained for the well-known autoassociation of insulin show the applicability of the method for the prediction of possible reaction modes between macromolecules.     

Semiquantitative analysis of rabbit knee articular surfaces based on stereomicroscopic examination of the cartilage

A semiquantitative stereomicroscopic method was devised in order to examine rabbit knee articular surfaces. With the aid of a drawing tube mounted on a stereomicroscope, enlarged pictures (magnification of X 14-19) of ink-stained or SEM specimens of joint surfaces were drawn and the structural details classified. The point-counting method or a computer-coupled analyzer was used to analyze the pictures. The data thus obtained underwent statistical evaluation. The method proved to be very useful for the quantitation of experimentally induced changes on cartilage surfaces.     

Using 96-well tissue culture polystyrene plates and a fluorescence plate reader as tools to study the survival and inactivation of viruses on surfaces

A method for studying the behavior of viruses on surfaces has been developed and is illustrated by determining the temperatures that inactivate adsorbed viral hemorrhagic septicemia virus (VHSV) and the concentration of 1-propanol that disinfected surfaces with adsorbed VHSV and chum salmon virus (CSV). VHSV is a rhabdovirus; CSV, a reovirus, and they were detected with two fish cell lines, EPC and CHSE-214, respectively. When polystyrene tissue culture surfaces were incubated with virus, rinsed, and left to dry, they still supported the attachment and spreading of cell lines and after 7 days these cells showed the characteristic CPE of the viruses. Thus cells appeared to be infected directly from surfaces on which viruses had been adsorbed. Applying this property to 96-well plates allowed duplicate surfaces to be examined for their infectiousness or support of CPE. For each treatment 80 replicate surfaces in a 96-well plate were tested at one time and the results expressed as the number of wells showing CPE. VHSV adsorbed to polystyrene was inactivated by drying in the dark at temperatures above 14 °C, but remained infectious for at least 15 days of drying at 4 °C. For chemical sterilization of polystyrene surfaces with adsorbed virus, disinfection was achieved with 1-propanol at 40% for VHSV and at 60% for CSV. As CPE can be conveniently monitored in 96-well plates with a fluorescence plate reader, this method can be used to rapidly evaluate a variety of treatments for their ability to inactivate surface-bound viruses.     

Terminally functionalized thermoresponsive polymer brushes for simultaneously promoting cell adhesion and cell sheet harvest

For preparing cell sheets effectively for cell sheet-based regenerative medicine, cell-adhesion strength to thermoresponsive cell culture surfaces need to be controlled precisely. To design new thermoresponsive surfaces via a terminal modification method, thermoresponsive polymer brush surfaces were fabricated through the surface-initiated reversible addition-fragmentation chain transfer (RAFT) radical polymerization of N-isopropylacrylamide (IPAAm) on glass substrates. The RAFT-mediated grafting method gave dithiobenzoate (DTB) groups to grafted PIPAAm termini, which can be converted to various functional groups. In this study, the terminal carboxylation of PIPAAm chains provided high cell adhesive property to thermoresponsive surfaces. Although cell adhesion is generally promoted by a decrease in the grafted PIPAAm amount, the decrease also decelerated thermally-induced cell detachment, whereas the influence of terminal modification was negligible on the cell detachment. Consequently, the terminally modified PIPAAm brush surfaces allowed smooth muscle cells (SMCs) to simultaneously adhere strongly and detach themselves rapidly. In this study, SMCs were unable to reach a confluent monolayer on as-prepared PIPAAm brush surfaces (grafted amount: 0.41 μg/cm(2)) without terminal carboxylation due to their insufficient cell-adhesion strength. On the other hand, though a decrease in the PIPAAm amount allowed SMCs to form a confluent cell monolayer on the PIPAAm brush surface, the SMCs were unable to be harvested as a monolithic cell sheet by low-temperature culture at 20 °C. Because of their unique property, only terminal-carboxylated PIPAAm brush surfaces achieved rapid harvesting of complete cell sheets by low-temperature culturing.     

Chemiluminescence-based detection and comparison of protein amounts adsorbed on differently modified silica surfaces

The biological consequences of protein adsorption on biomaterial surfaces are considered to be of utmost importance for their biocompatibility. A new method based on amino group-labeling coupled to a chemiluminescence reaction for direct determination of proteins adsorbed on material surfaces was employed. This method was used to explore the effects of surface chemistry and surface roughness on protein adsorption in a silicon oxide model system. Corundum sandblasting was applied to silicon wafers to create roughened surfaces while immobilization of fluorocarbon-, hydrocarbon-, and poly(ethylene glycol)-containing silanes produced surfaces of varying wettability. The adsorption behavior of two complex body fluids, human serum and saliva, and of two purified components, human serum albumin and fibronectin, was strongly influenced by the surface parameters. A general tendency to higher amounts of adsorbed protein was found on roughened surfaces and modification with poly(ethylene glycol) or with fluorocarbon moieties reduced protein adsorption. The values obtained with the new method could be confirmed by a colorimetric determination of protein amounts adsorbed on identically modified silica beads and were in accordance with those previously reported utilizing established methods for protein quantification. The presented method, which was methodically simple to perform and allowed the simultaneous measurement of a large number of samples, may be of future value for high-throughput surveying of the protein adsorption characteristics of biomaterials.     

Matrix-assisted laser desorption ionization mass spectrometry: a new tool for probing interactions between proteins and metal surfaces. Use in dental implantology.

The fixation in the bone of an artificial titanium tooth root is believed to be initiated by the rapid adsorption of the proteins present in the surgical cavity on the titanium surface. The study of this adsorption should make it possible to predict the osseointegration capacities of new implant surface treatments. We describe here a new method, based on matrix-assisted laser desorption ionization-mass spectrometry (MALDI-MS), for quantifying proteins adsorbed on titanium surfaces fully identical to these designed for implantology. The key step of this method is a new MALDI-MS sample preparation allowing the adsorbed proteins to be removed from the surface and to be homogeneously dispersed in the matrix crystals. The adsorption of a model protein (lysozyme) on two titanium surfaces (polished and sandblasted) was studied in order to evaluate the method. The absolute MALDI-MS intensity was shown to vary linearly with the amount of adsorbed lysozyme. After dipping the titanium surfaces for different times in lysozyme solutions at different concentrations, the maximum amount of adsorbed lysozyme was measured by MALDI-MS and was shown to correspond to a lysozyme monolayer, which is consistent with results described in the literature.     

大尺度不透水面遥感估算方法比较——以京津唐为例

城市不透水面既是常用的城市化程度指标,也是衡量环境质量的重要指标。采用遥感技术准确提取城市不透水面并分析其空间扩张过程,对生态城市建设具有重要意义。基于Landsat 5 TM影像,采用NDVI二元法和线性光谱分解法,分别提取北京、天津和唐山3个城市不透水面信息,并将不透水面估算结果与近同期的ALOS影像提取结果对比验证。结果表明,线性光谱分解法获取的不透水面结果较好,RMSE为20.6%,能有效提取大范围的不透水面信息。     

Droplets motion on chemically/topographically heterogeneous surfaces

We present a numerical study of droplets sliding across chemically heterogeneous surfaces formed by a periodic pattern of alternating hydrophobic and hydrophilic stripes, or topographically heterogeneous surfaces which are microgrooved. The numerical simulation performed by using a particle-based numerical method, Many-body Dissipative Particle Dynamics (MDPD), is adopted to observe the stick-slip motion of droplets driven by a constant body force. The fractions of two types of surfaces are varied from 0.3 to 0.7 to investigate their influence on stick-slip motion of droplets. The dynamic contact angles and the variation in distance D_fr between the front and the rear contact points are shown for different fractions. The jerky motion characterised by stick-slip motion can be found on chemically heterogeneous surfaces for all fractions we choose and the partial stick-slip motion can also be discovered on topographically heterogeneous surfaces except for fraction Φ_s = 0.3. The snapshots of droplet show that stick-slip motion is the consequence of the periodic deformation of droplet interface during crossing heterogeneous surfaces and can be controlled by fractions.     

Sampling with adhesive tape strips: an easy and rapid method to monitor microbial colonization on monument surfaces

An easy and fast non-destructive method for sampling from monument and art object surfaces is proposed. The results obtained after sampling in regions of black spots and discoloration using adhesive tape strips showed that the method is useful for monitoring microbial colonization as well as for the identification of biodeteriogens. This technique is easy to apply, inexpensive and reproducible. The presence of fungi or algae on stone surfaces can be checked at an early stage of colonization or after cleaning procedures, without damage to the sampled surface. In addition, it is possible to obtain information on the morphology and taxonomy of microorganisms, and their relationships with the colonized material surfaces.     

A new approach for surface fitting method of articular joint surfaces

The application of joint contact mechanics requires a precise configuration of the joint surfaces. B-Spline, and NURBS have been widely used to model joint surfaces, but because these formulations use a structured data set provided by a rectangular net first, then a grid, there is a limit to the accuracy of the models they can produce. However new imaging systems such as 3D laser scanners can provide more realistic unstructured data sets. What is needed is a method to manipulate the unstructured data. We created a parametric polynomial function and applied it to unstructured data sets obtained by scanning joint surfaces. We applied our polynomial model to unstructured data sets from an artificial joint, and confirmed that our polynomial produced a smoother and more accurate model than the conventional B-spline method. Next, we applied it to a diarthrodial joint surface containing many ripples, and found that our function's noise filtering characteristics smoothed out existing ripples. Since no formulation was found to be optimal for all applications, we used two formulations to model surfaces with ripples. First, we used our polynomial to describe the global shape of the objective surface. Minute undulations were then specifically approximated with a Fourier series function. Finally, both approximated surfaces were superimposed to reproduce the original surface in a complete fashion.     

Label-fracture of cell surfaces by replica staining

We introduce replica-staining label-fracture, a method for the cytochemical mapping of membrane surfaces. This method is a corollary of the rationale of label-fracture (Pinto da Silva and Kan, 1984: J Cell Biol 99:1156). After freeze-fracture the exoplasmic halves of the membrane remain attached to the replica. We show that cytochemical labeling of cell surfaces can be performed by direct post-fracture staining of freeze-fracture replicas. This new variant of label-fracture leads to miniaturization of labeling procedures and allows standardization of labeling conditions and simultaneous processing of different specimens.     

Fitting parametrized polynomials with scattered surface data.

Currently used joint-surface models require the measurements to be structured according to a grid. With the currently available tracking devices a large quantity of unstructured surface points can be measured in a relatively short time. In this paper a method is presented to fit polynomial functions to three-dimensional unstructured data points. To test the method spherical, cylindrical, parabolic, hyperbolic, exponential, logarithmic, and sellar surfaces with different undulations were used. The resulting polynomials were compared with the original shapes. The results show that even complex joint surfaces can be modelled with polynomial functions. In addition, the influence of noise and the number of data points was also analyzed. From a surface (diam: 20 mm) which is measured with a precision of 0.2 mm a model can be constructed with a precision of 0.02 mm.     

Thermoresponsive protein adsorption of poly(N-isopropylacrylamide)-modified streptavidin on polydimethylsiloxane microchannel surfaces

The control of protein adsorption on microchannel surfaces is important for biosensors. In this study, we demonstrated protein adsorption method that is controlled through temperature change, i.e., thermoresponsive protein adsorption, on polydimethylsiloxane (PDMS) microchannel surfaces using a thermoresponsive polymer, poly(N-isopropylacrylamide) (PNIPAAm). To provide general protein adsorption control method, we adopted biotin-streptavidin chemistry and synthesized streptavidin covalently modified with PNIPAAm (PNIPAAm-StAv). Modification of streptavidin, a hydrophilic protein, with PNIPAAm induced successful thermoresponsive adsorption on a PDMS microchannel surfaces: PNIPAAm-StAv adsorbed at 37 degrees C and desorbed at 10 degrees C on the surfaces. We also demonstrated the thermoresponsive adsorption of biotinylated immunoglobulin G (IgG-b) using PNIPAAm-StAv. Conjugation of IgG-b with PNIPAAm-StAv induced successful thermoresponsive IgG-b adsorption on PDMS. Modification of PDMS surfaces with PNIPAAm reduced physical adsorption of the partially hydrophobic IgG-b on the surface and contributed to the high-contrast thermoresponsive adsorption of IgG-b: less than 1% of the IgG-b adsorbed at 37 degrees C was detected after the PNIPAAm-PDMS surface was washed at 10 degrees C. The controllable adsorption of this system is expected to be applied to the regeneration of biosensor chips and to on-chip protein manipulation.     

Estimating surface growth rates from changes in curvature

The shape of a biological surface may be regarded as an observable. Here a method is given for deriving growth parameters from the change in shape of such a surface. Isotropy is assumed, and implies a conformal relationship between initial and final surfaces. One further assumption is necessary to specify the growth regime: in the case of radially symmetric surfaces, this is that the process is similarly symmetric; in the general case the assumption is that the Dirichlet integral of scale factors is miminized.     

Yeast transformation process studied by fluorescence labeling technique

A new method based on fluorescence imaging and flow cytometry was developed to investigate the transformation process of Saccharomyces cerevisiae AY. Yeast and fluorescent-labeled plasmid pUC18 were used as models of cells and DNA molecules, respectively. Binding of DNA molecules to yeast cell surfaces was observed. Factors influencing DNA binding to cell surfaces were investigated. It has been found that poly(ethylene glycol) (PEG) could induce DNA binding to yeast surfaces, while Li(+) showed a weak effect on the binding. When both Li(+) and PEG were used, synergetic effect occurred, resulting in the binding of pUC18 to the surface of more yeast cells compared with that in the presence of PEG or Li(+) only. It was also confirmed that heat shock, Li(+), and PEG all can increase the permeability of yeast cells. This simple method is helpful for understanding the process of yeast transformation and can be used to investigate the interaction of DNA with cell surfaces.