Catherine M. Klapperich

Nanomechanical Properties of Polymers Determined From Nanoindentation Experiments

The nanomechanical properties of various polymers were examined in light of nanoindentation experiments performed with a diamond tip of nominal radius of curvature of about 20 μm under conditions of maximum contact load in the range of 150-600 μN and loading/unloading rates between 7.5 and 600 μN/s. The elastic modulus of each polymer was determined from the unloading material response using the compliance method, whereas the hardness was calculated as the maximum contact load divided by the corre-sponding projected area, obtained from the known tip shape function. It is shown that while the elastic modulus decreases with increasing indentation depth, the polymer hard-ness tends to increase, especially for the polymers possessing amorphous microstructures or less crystallinity. Differences in the material properties, surface adhesion, and time-dependent deformation behavior are interpreted in terms of the microstructure, crystallinity, and surface chemical state of the polymers. Results obtained at different maximum loads and loading rates demonstrate that the nanoindentation technique is an effective method of differentiating the mechanical behavior of polymeric materials with different microstructures.

An integrated disposable device for DNA extraction and helicase dependent amplification

Here we report the demonstration of an integrated microfluidic chip that performs helicase dependent amplification (HDA) on samples containing live bacteria. Combined chip-based sample preparation and isothermal amplification are attractive for world health applications, since the need for instrumentation to control flow rate and temperature changes are reduced or eliminated. Bacteria lysis, nucleic acid extraction, and DNA amplification with a fluorescent reporter are incorporated into a disposable polymer cartridge format. Smart passive fluidic control using a flap valve and a hydrophobic vent (with a nanoporous PTFE membrane) with a simple on-chip mixer eliminates multiple user operations. The device is able to detect as few as ten colony forming units (CFU) of E. coli in growth medium.

Mechanical and chemical analysis of plasma and ultraviolet–ozone surface treatments for thermal bonding of polymeric microfluidic devices

Here we have demonstrated that radio frequency plasma and ultraviolet-ozone (UVO) surface modifications are effective treatments for enabling the thermal bonding of polymeric microfluidic chips at temperatures below the T(g) (glass transition temperature) of the polymer. The effects of UVO and plasma treatments on the surface properties of a cyclic polyolefin and polystyrene were examined with X-ray photoelectron spectroscopy (XPS), contact angle measurements, atomic force microscopy (AFM) surface roughness measurements and surface adhesion measurements with AFM force-distance data. Three-point bending tests using a dynamic mechanical analyzer (DMA) were used to characterize the bond strength of thermally sealed polymer parts and the cross-sections of the bonded microchannels were evaluated with scanning electron microscopy (SEM). The experimental results demonstrated that plasma and UVO surface treatments cause changes in the chemical and physical characteristics of the polymer surfaces, resulting in a decrease in T(g) at the surface, and thus allowing the microfluidic chips to be effectively bonded at temperatures lower than the T(g) of the bulk polymer without losing the intended channel geometry.

Time-dependent mechanical characterization of poly(2-hydroxyethyl methacrylate) hydrogels using nanoindentation and unconfined compression.

Hydrogels pose unique challenges to nanoindentation including sample preparation, control of experimental parameters, and limitations imposed by mechanical testing instruments and data analysis originally intended for harder materials. The artifacts that occur during nanoindentation of hydrated samples have been described, but the material properties obtained from hydrated nanoindentation have not yet been related to the material properties obtained from macroscale testing. To evaluate the best method for correlating results from microscale and macroscale tests of soft materials, nanoindentation and unconfined compression stress-relaxation tests were performed on poly-2-hydroxyethyl methacrylate (pHEMA) hydrogels with a range of cross-linker concentrations. The nanoindentation data were analyzed with the Oliver-Pharr elastic model and the Maxwell-Wiechert (j = 2) viscoelastic model. The unconfined compression data were analyzed with the Maxwell-Wiechert model. This viscoelastic model provided an excellent fit for the stress-relaxation curves from both tests. The time constants from nanoindentation and unconfined compression were significantly different, and we propose that these differences are due to differences in equilibration time between the microscale and macroscale experiments and in sample geometry. The Maxwell-Wiechert equilibrium modulus provided the best agreement between nanoindentation and unconfined compression. Also, both nanoindentation analyses showed an increase in modulus with each increasing cross-linker concentration, validating that nanoindentation can discriminate between similar, low-modulus, hydrated samples.

Surface detection errors cause overestimation of the modulus in nanoindentation on soft materials.

The accuracy of mechanical properties from depth-sensing indentation, or nanoindentation, depends on the accuracy of the displacement measurement used to calculate these properties. Here, current nanoindentation techniques and analysis methods for accurate displacement measurements are reviewed. First, the ability of a commercial instrument to sense the surface of soft materials is examined. Second, methods of sample surface detection are reviewed. Finally, a case of overestimation of the elastic modulus of a compliant material using nanoindentation with incorrect displacement values is presented.

Clinically relevant microfluidic magnetophoretic isolation of rare-cell populations for diagnostic and therapeutic monitoring applications.

Cells of biomedical interest are, despite their functional significance, often present in very small numbers. Therefore the analysis and isolation of previously inaccessible rare cells, such as peripheral hematopoietic stem cells, endothelial progenitor cells, or circulating tumor cells, require efficient, sensitive, and specific procedures that do not compromise the viability of the cells. The current study builds on previous work on a rationally designed microfluidic magnetophoretic cell separation platform capable of throughputs of 240 μL min(-1). Proof-of-concept was first conducted using MCF-7 (1-1000 total cells) as the target rare cell spiked into high concentrations of Raji B-lymphocyte nontarget cells (~10(6) total cells). These experiments lead to the establishment of a magnet-based separation for the isolation of 50 MCF-7 cells directly from whole blood. Results show an efficiency of collection greater than 85%, with a purity of over 90%. Next, resident endothelial progenitor cells and hematopoietic stem cells are directly isolated from whole human blood in a rapid and efficient fashion (>96%). Both cell populations could be simultaneously isolated and, via immunofluorescent staining, individually identified and enumerated. Overall, the presented device illustrates a viable separation platform for high purity, efficient, and rapid collection of rare cell populations directly from whole blood samples.

Failure of a metal-on-metal total hip arthroplasty from progressive osteolysis☆

Ultra-high-molecular weight polyethylene (UHMWPE) wear, debris-induced osteolysis is a frequent cause of failure of total hip arthroplasty. Metal-on-metal total hip arthroplasty eliminates the generation of UHMWPE particulate debris. Although the volumetric wear of a metal-on-metal articulation may be lower than a metal-UHMWPE articulation, the number of particles may be higher. Osteolysis can develop in response to metallic and UHMWPE debris. The following case of massive osteolysis associated with large amounts of cobalt-chrome wear debris shows adhesive and abrasive wear mechanisms, as well as wear caused by third-body cobalt-chrome debris and impingement of the femoral component against the rim of the acetabular cup, which led to failure of a metal-on-metal total hip arthroplasty.

Rapid point-of-care concentration of bacteria in a disposable microfluidic device using meniscus dragging effect

We report a low cost, disposable polymer microfluidic sample preparation device to perform rapid concentration of bacteria from liquid samples using enhanced evaporation targeted at downstream detection using surface enhanced Raman spectroscopy (SERS). The device is composed of a poly(dimethylsiloxane) (PDMS) liquid sample flow layer, a reusable metal airflow layer, and a porous PTFE (Teflon™) membrane sandwiched in between the liquid and air layers. The concentration capacity of the device was successfully demonstrated with fluorescently tagged Escherichia coli (E. coli). The recovery concentration was above 85% for all initial concentrations lower than 1 × 10(4) CFU mL(-1). In the lowest initial concentration cases, 100 µL initial volumes of bacteria solution at 100 CFU mL(-1) were concentrated into 500 nL droplets with greater than 90% efficiency in 15 min. Subsequent tests with SERS on clinically relevant Methicillin-Sensitive Staphylococcus aureus (MSSA) after concentration in this device proved more than 100-fold enhancement in SERS signal intensity compared to the signal obtained from the unconcentrated sample. The concentration device is straightforward to design and use, and as such could be used in conjunction with a number of detection technologies.

Comparison of three joint simulator wear debris isolation techniques: acid digestion, base digestion, and enzyme cleavage.

Quantification of ultrahigh molecular weight polyethylene (UHMWPE) wear debris remains a challenging task in orthopedic device analysis. Currently, the weight loss method is the only accepted practice for quantifying the amount of wear generated from a PE component. This technique utilizes loaded soak controls and weight differences to account for polymeric material lost through wear mechanisms. This method enables the determination of the amount of wear in the orthopedic device, but it provides no information about debris particulate size distribution. In order to shed light on wear mechanisms, information about the wear debris and its size distribution is necessary. To date, particulate isolation has been performed using the base digestion technique. The method uses a strong base, ultracentrifugation, and filtration to digest serum constituents and to isolate PE debris from sera. It should be noted that particulate isolation methods provide valuable information about particulate size distribution and may elucidate the mechanisms of wear associated with polymeric orthopedic implants; however, these techniques do not yet provide a direct measure of the amount of wear. The aim of this study is to present alternative approaches to wear particle isolation for analysis of polymer wear in total joint replacements without recourse to ultracentrifugation. Three polymer wear debris isolation techniques (the base method, an acid treatment, and an enzymatic digestion technique) are compared for effectiveness in simulator studies. A requirement of each technique is that the wear particulate must be completely devoid of serum proteins in order to effectively image and count these particles. In all methods the isolation is performed through filtration and chemical treatment. Subsequently, the isolated polymer particles are imaged using scanning electron microscopy and quantified with digital image analysis. The results from this study clearly show that isolation can be performed without the use of ultracentrifugation and that these methods provide a viable option for wear debris analysis.

Tribological Properties and Microstructure Evolution of Ultra-High Molecular Weight Polyethylene

The friction and wear properties of unmodified ultra-high molecular weight polyethylene (UHMWPE) were investigated experimentally. Disks of semicrystalline UHMWPE were slid against polished CoCrWNi pins in bovine serum at ranges of contact pressure and sliding speed typical of those encountered in total joint replacements. The coefficient of friction was monitored continuously during testing, and the wear rate was determined from surface profilometry measurements of worn disk surfaces accounting for strain relaxation. Scanning electron microscopy (SEM) results demonstrated that surface deterioration comprises adhesion, third-body abrasion by polyethylene wear debris, and delamination wear. The contribution of these mechanisms to the overall wear rate and the formation of wear debris depends predominantly on the contact pressure and secondarily on the sliding speed. Transmission electron microscopy (TEM) yielded new insight into the evolution of the microstructure morphology of UHMWPE during sliding. Cross sections parallel to the wear tracks obtained from various depths were analyzed with the TEM to develop a spatial mapping of the subsurface microstructure as a function of contact pressure. Alignment of crystalline regions (lamellae) in the polyethylene microstructure parallel to the sliding surface was found to occur during sliding even at relatively low contact pressures. SEM observations suggested that the highly oriented microstructure is the precursor to delamination wear, leading to the formation of wear particles larger than those produced by adhesion and third-body abrasion at the contact interface.