A methodological framework for nanomechanical characterization of soft biomaterials and polymers.

Nanoindentation utilizes a hard indenter probe to deform the sample surface in order to measure local properties, such as indentation modulus and hardness. Initially intended for characterization of elastic and elastic-plastic materials, nanoindentation has more recently been utilized for viscoelastic solids as well as hydrated and soft biological materials. An advantage to nanoindentation is the ability to determine the nano- and microscale properties of materials with complex microstructures as well as those of limited sample dimension. Nanoindentation finds utility in the characterization of structural tissues, hydrogels, polymers and composites. Nevertheless, testing complexities such as adhesion and surface detection exist in nanoindentation of compliant viscoelastic solids and hydrated materials. These challenges require appropriate modifications in methodology and use of appropriate contact models to analyze nanoindentation data. A full discussion of protocol adjustments has yet to be assembled into a robust nanoindentation testing framework of soft biomaterials and polymers. We utilize existing nanoindentation literature and testing expertise in our laboratories to (1) address challenges and potential errors when performing indentations on soft or hydrated materials, (2) explore best practices for mitigating experimental error, and (3) develop a nanoindentation framework that serves researchers as a primer for nanoindentation testing of soft/hydrated biomaterials and polymers.

A 3D-transient elastohydrodynamic lubrication hip implant model to compare ultra high molecular weight polyethylene with more compliant polycarbonate polyurethane acetabular cups.

Wear remains a significant challenge in the design of orthopedic implants such as total hip replacements. Early elastohydrodynamic lubrication modeling has predicted thicker lubrication films in hip replacement designs with compliant polycarbonate polyurethane (PCU) bearing materials compared to stiffer materials like ultra-high molecular weight polyethylene (UHMWPE). The predicted thicker lubrication films suggest improved friction and wear performance. However, when compared to the model predictions, experimental wear studies showed mixed results. The mismatch between the model and experimental results may lie in the simplifying assumptions of the early models such as: steady state conditions, one dimensional rotation and loading, and high viscosities. This study applies a 3D-transient elastohydrodynamic model based on an ISO standard gait cycle to better understand the interaction between material stiffness and film thickness in total hip arthroplasty material couples. Similar to previous, simplified models, we show that the average and central film thickness of PCU (∼0.4µm) is higher than that of UHMWPE (∼0.2µm). However, in the 3D-transient model, the film thickness distribution was largely asymmetric and the minimum film thickness occurred outside of the central axis. Although the overall film thickness of PCU was higher than UHMWPE, the minimum film thickness of PCU was lower than UHMPWE for the majority of the gait cycle. The minimum film thickness of PCU also had a larger range throughout the gait cycle. Both materials were found to be operating between boundary and mixed lubrication regimes. This 3D-transient model reveals a more nuanced interaction between bearing material stiffness and film thickness that supports the mixed results found in experimental wear studies of PCU hip implant designs.

Oxidized Zirconium Components Maintain a Smooth Articular Surface Except Following Hip Dislocation.

BACKGROUND: Oxidized zirconium (OxZr) offers theoretical advantages in total hip and knee arthroplasty (THA and TKA, respectively) relative to other biomaterials by combining the tribological benefits of ceramics with the fracture toughness of metals. Yet, some studies have found that OxZr does not improve outcomes or wear rates relative to traditional bearing materials such as cobalt-chromium (CoCr). Separately, effacement of the thin ceramic surface layer has been reported for OxZr components, though the prevalence and sequelae are unclear. METHODS: To elucidate the in vivo behavior of OxZr implants, the articular surfaces of 94 retrieved THA and TKA femoral components (43 OxZr TKA, 21 OxZr THA, 30 CoCr THA) were analyzed using optical microscopy, non-contact profilometry, and scanning electron microscopy. RESULTS: We found that OxZr components maintain a smooth articular surface except following hip dislocation. Three of four OxZr femoral heads revised following dislocation exhibited severe damage to the articular surface, including macroscopic regions of ceramic-layer effacement and exposure of the underlying metal substrate; these components were 23-32 times rougher than pristine OxZr controls. When revised for dislocation, OxZr femoral heads were substantially rougher than CoCr femoral heads (median Sa = 0.431 v. 0.020 µm, P = .03). In contrast, CoCr femoral heads exhibited low overall roughness values regardless of whether they dislocated (median Sa = 0.020 v. 0.008 µm, P = .09, CoCr dislocators v. non-dislocators). CONCLUSIONS: Effacement of the ceramic surface layer and substantial articular surface roughening is not atypical following dislocation of OxZr femoral heads, making OxZr much less tolerant than CoCr to hip dislocation.

Nanomechanical analysis of medical grade PEEK and carbon fiber-reinforced PEEK composites.

Polyether ether ketone (PEEK) and PEEK composites are viable candidates for orthopedic implants owing to their ability for modulus match of surrounding bone tissue. The structural properties of these systems for load-bearing application in the body can be tailored by incorporating carbon fibers; to this end, polyacrylonitrile (PAN) and pitch fibers are commonly incorporated in the PEEK matrix. Mechanical property optimization for a given medical application requires consideration of carbon fiber type and volume fraction, as well as processing conditions for the composite systems. While much is known about the bulk mechanical properties of PEEK and PEEK composites, little is known about the nanomechanical properties of these systems. Insight into nanoscale behavior can offer valuable information about fiber-matrix interactions that may influence long-term integrity of these biomaterials when used in load bearing medical device applications. In this study, we utilize nanoindentation as a method to characterize mechanical behavior of clinical grade PEEK and PEEK composites. We examine PEEK formulations with pitch and PAN fibers and evaluate a range of thermal treatments known to influence polymer microstructure. We use a conospherical tip of 1.5 µm in radius and a conospherical tip of 20 µm radius to determine indentation modulus over different length scales. We correlate these findings with previous characterization on these same PEEK systems using microindentation. A novelty of this work is that we combine nanoindentation with k-means clustering to quantitatively discern the influence of heat treatment and carbon fiber type on the mechanical behavior of PEEK composites and their constituents. We demonstrate that nanoindentation is an effective characterization tool for discerning fiber-matrix interactions and measuring the mechanical behavior in response to thermal treatment and carbon fiber type in PEEK composites. Nanoindentation is shown to be a viable tool for characterizing complex biomaterials and can serve as an effective technique to guide optimization of microstructures for long-term structural applications in the body.

Micromechanisms of fatigue crack growth in polycarbonate polyurethane: Time dependent and hydration effects.

Polycarbonate polyurethane has cartilage-like, hygroscopic, and elastomeric properties that make it an attractive material for orthopedic joint replacement application. However, little data exists on the cyclic loading and fracture behavior of polycarbonate polyurethane. This study investigates the mechanisms of fatigue crack growth in polycarbonate polyurethane with respect to time dependent effects and conditioning. We studied two commercially available polycarbonate polyurethanes, Bionate® 75D and 80A. Tension testing was performed on specimens at variable time points after being removed from hydration and variable strain rates. Fatigue crack propagation characterized three aspects of loading. Study 1 investigated the impact of continuous loading (24h/day) versus intermittent loading (8-10h/day) allowing for relaxation overnight. Study 2 evaluated the effect of frequency and study 3 examined the impact of hydration on the fatigue crack propagation in polycarbonate polyurethane. Samples loaded intermittently failed instantaneously and prematurely upon reloading while samples loaded continuously sustained longer stable cracks. Crack growth for samples tested at 2 and 5Hz was largely planar with little crack deflection. However, samples tested at 10Hz showed high degrees of crack tip deflection and multiple crack fronts. Crack growth in hydrated samples proceeded with much greater ductile crack mouth opening displacement than dry samples. An understanding of the failure mechanisms of this polymer is important to assess the long-term structural integrity of this material for use in load-bearing orthopedic implant applications.

Notch fatigue of ultrahigh molecular weight polyethylene (UHMWPE) used in total joint replacements.

Ultrahigh molecular weight polyethylene (UHMWPE) has remained the primary polymer used in hip, knee and shoulder replacements for over 50 years. Recent case studies have demonstrated that catastrophic fatigue fracture of the polymer can severely limit device lifetime and are often associated with stress concentration (notches) integrated into the design. This study evaluates the influence of notch geometry on the fatigue of three formulations of UHMWPE that are in use today. A linear-elastic fracture mechanics approach is adopted to evaluate crack propagation as a function of notch root radius, heat treatment and Vitamin E additions. Specifically, a modified stress-intensity factor that accounts for notch geometry was utilized to model the crack driving force. The degree of notch plasticity for each material/notch combination was further evaluated using finite element methods. Experimental evaluation of crack speed as a function of stress intensity was conducted under cyclic tensile loading, taking crack length and notch plasticity into consideration. Results demonstrated that crack propagation in UHMWPE emanating from a notch was primarily affected by microstructural influences (cross-linking) rather than differences in notch geometry.

Effects of elastase and collagenase on the nonlinearity and anisotropy of porcine aorta.

We use enzymatic manipulation methods to investigate the individual and combined roles of elastin and collagen on arterial mechanics. Porcine aortic tissues were treated for differing amounts of time using enzymes elastase and collagenase to cause degradation in substrate proteins elastin and collagen and obtain variable tissue architecture. We use equibiaxial mechanical tests to quantify the material properties of control and enzyme treated tissues and histological methods to visualize the underlying tissue microstructure in arterial tissues. Our results show that collagenase treated tissues were more compliant in the longitudinal direction as compared to control tissues. Collagenase treatment also caused a decrease in the tissue nonlinearity as compared to the control samples in the study. A one hour collagenase treatment was sufficient to cause fragmentation and degradation of the adventitial collagen. In contrast, elastase treatment leads to significantly stiffer tissue response associated with fragmented and incomplete elastin networks in the tissue. Thus, elastin in arterial walls distributes tensile stresses whereas collagen serves to reinforce the vessel wall in the circumferential direction and also contributes to tissue anisotropy. A microstructurally motivated strain energy function based on circumferentially oriented medial fibers and helically oriented collagen fibers in the adventitia is useful in describing these experimental results.

Clinical trade-offs in cross-linked ultrahigh-molecular-weight polyethylene used in total joint arthroplasty.

Highly cross-linked formulations of ultrahigh-molecular-weight polyethylene (XLPE) offer exceptional wear resistance for total joint arthroplasty but are offset with a reduction in postyield and fatigue fracture properties in comparison to conventional ultrahigh-molecular-weight polyethylene (UHMWPE). Oxidation resistance is also an important property for the longevity of total joint replacements (TJRs) as formulations of UHMWPE or XLPE utilizing radiation methods are susceptible to free radical generation and subsequent embrittlement. The balance of oxidation, wear, and fracture properties is an enduring concern for orthopedic polymers used as the bearing surface in total joint arthroplasty. Optimization of material properties is further challenged in designs that make use of locking mechanisms, notches, or other stress concentrations that can render the polymer susceptible to fracture due to elevated local stresses. Clinical complications involving impingements, dislocations, or other biomechanical overloads can exacerbate stresses and negate benefits of improved wear resistance provided by XLPE. This work examines trade-offs that factor into the use of XLPE in TJR implants.

The effect of E-glass fibers and acrylic resin thickness on fracture load in a simulated implant-supported overdenture prosthesis.

STATEMENT OF PROBLEM: Implant overdenture prostheses are prone to acrylic resin fracture because of space limitations around the implant overdenture components. PURPOSE: The purpose of this study was to evaluate the influence of E-glass fibers and acrylic resin thickness in resisting acrylic resin fracture around a simulated overdenture abutment. MATERIAL AND METHODS: A model was developed to simulate the clinical situation of an implant overdenture abutment with varying acrylic resin thickness (1.5 or 3.0 mm) with or without E-glass fiber reinforcement. Forty-eight specimens with an underlying simulated abutment were divided into 4 groups (n=12): 1.5 mm acrylic resin without E-glass fibers identified as thin with no E-glass fiber mesh (TN-N); 1.5 mm acrylic resin with E-glass fibers identified as thin with E-glass fiber mesh (TN-F); 3.0 mm acrylic resin without E-glass fibers identified as thick without E-glass fiber mesh (TK-N); and 3.0 mm acrylic resin with E-glass fibers identified as thick with E-glass fiber mesh (TK-F). All specimens were submitted to a 3-point bending test and fracture loads (N) were analyzed with a 2-way ANOVA and Tukey's post hoc test (α=.05). RESULTS: The results revealed significant differences in fracture load among the 4 groups, with significant effects from both thickness (P<.001) and inclusion of the mesh (P<.001). Results demonstrated no interaction between mesh and thickness (P=.690). The TN-N: 39 ±5 N; TN-F: 50 ±6.9 N; TK-N: 162 ±13 N; and TK-F: 193 ±21 N groups were all statistically different (P<.001). CONCLUSIONS: The fracture load of a processed, acrylic resin implant-supported overdenture can be significantly increased by the addition of E-glass fibers even when using thin acrylic resin sections. On a relative basis, the increase in fracture load was similar when adding E-glass fibers or increasing acrylic resin thickness.

Tradeoffs amongst fatigue, wear, and oxidation resistance of cross-linked ultra-high molecular weight polyethylene.

This study evaluated the tradeoffs amongst fatigue crack propagation resistance, wear resistance, and oxidative stability in a wide variety of clinically-relevant cross-linked ultra-high molecular weight polyethylene. Highly cross-linked re-melted materials showed good oxidation and wear performance, but diminished fatigue crack propagation resistance. Highly cross-linked annealed materials showed good wear and fatigue performance, but poor oxidation resistance. Moderately cross-linked re-melted materials showed good oxidation resistance, but moderate wear and fatigue resistance. Increasing radiation dose increased wear resistance but decreased fatigue crack propagation resistance. Annealing reduced fatigue resistance less than re-melting, but left materials susceptible to oxidation. This appears to occur because annealing below the melting temperature after cross-linking increased the volume fraction and size of lamellae, but failed to neutralize all free radicals. Alternately, re-melting after cross-linking appeared to eliminate free radicals, but, restricted by the network of cross-links, the re-formed lamellae were fewer and smaller in size which resulted in poor fatigue crack propagation resistance. This is the first study to simultaneously evaluate fatigue crack propagation, wear, oxidation, and microstructure in a wide variety of clinically-relevant ultra-high. The tradeoff we have shown in fatigue, wear, and oxidation performance is critical to the material's long-term success in total joint replacements.

The use of polyacrylamide gels for mechanical calibration of cartilage--a combined nanoindentation and unconfined compression study.

This study investigates polyacrylamide (PA) gel as a calibration material to measure the nanomechanical compressive modulus of cartilage using nanoindentation. Both nanoindentation and unconfined compression testing were performed on PA gel and porcine rib cartilage. The equilibrium moduli measured by the two methods were discernable. Nanoindentation has the advantage of distinguishing between spatially dependent constituent properties that affect tissue mechanical function in heterogeneous and hierarchically structured tissues such as cartilage. Both sets of measurements exhibited similar positive correlation with increasing gel crosslinker concentration. The compressive modulus measurements from compression in the PA gels ranged from 300 kPa-1.4 MPa, whereas those from nanoindentation ranged from 100 kPa-1.1 MPa. Using this data, a method for relating nanoindentation measurements to conventional mechanical property measurements is presented for porcine rib cartilage. It is shown that based on this relationship, the local tissue modulus as measured from nanoindentation (1.1-1.4 MPa) was able to predict the overall global modulus of the same sample of rib cartilage (2.2 MPa), as confirmed by experimental measurements from unconfined compression. This study supports the use of nanoindentation for the local characterization of cartilage tissues and may be applied to other soft tissues and constructs.

Microscale wear behavior and crosslinking of PEG-like coatings for total hip replacements.

The predominant cause of late-state failure of total hip replacements is wear-mediated osteolysis caused by wear particles that originate from the ultrahigh molecular weight polyethylene (UHMWPE) acetabular cup surface. One strategy for reducing wear particle formation from UHMWPE is to modify the surface with a hydrophilic coating to increase lubrication from synovial fluid. This study focuses on the wear behavior of hydrophilic coatings similar to poly(ethylene glycol) (PEG). The coatings were produced by plasma-polymerizing tetraglyme on UHMWPE in a chamber heated to 40 degrees C or 50 degrees C. Both temperatures yielded coatings with PEG-like chemistry and increased hydrophilicity relative to uncoated UHMWPE; however, the 40 degrees C coatings were significantly more resistant to damage induced by atomic force microscopy nanoscratching. The 40 degrees C coatings exhibited only one damage mode (delamination) and often showed no signs of damage after repeated scratching. In contrast, the 50 degrees C coatings exhibited three damage modes (roughening, thinning, and delamination), and always showed visible signs of damage after no more than two scratches. The greater wear resistance of the 40 degrees C coatings could not be explained by coating chemistry or hydrophilicity, but it corresponded to an approximately 26-32% greater degree of crosslinking relative to the 50 degrees C surfaces, suggesting that crosslinking should be a significant design consideration for hydrophilic coatings used for total hip replacements and other wear-dependent applications.

Characterization and tribology of PEG-like coatings on UHMWPE for total hip replacements.

A crosslinked hydrogel coating similar to poly(ethylene glycol) (PEG) was covalently bonded to the surface of ultrahigh molecular weight polyethylene (UHMWPE) to improve the lubricity and wear resistance of the UHWMPE for use in total joint replacements. The chemistry, hydrophilicity, and protein adsorption resistance of the coatings were determined, and the wear behavior of the PEG-like coating was examined by two methods: pin-on-disk tribometry to evaluate macroscale behavior, and atomic force microscopy (AFM) to simulate asperity wear. As expected, the coating was found to be highly PEG-like, with approximately 83% ether content by x-ray photoelectron spectroscopy and more hydrophilic and resistant to protein adsorption than uncoated UHMWPE. Pin-on-disk testing showed that the PEG-like coating could survive 3 MPa of contact pressure, comparable to that experienced by total hip replacements. AFM nanoscratching experiments uncovered three damage mechanisms for the coatings: adhesion/microfracture, pure adhesion, and delamination. The latter two mechanisms appear to correlate well with wear patterns induced by pin-on-disk testing and evaluated by attenuated total reflection Fourier transform infrared spectroscopy mapping. Understanding the mechanisms by which the PEG-like coatings wear is critical for improving the behavior of subsequent generations of wear-resistant hydrogel coatings.

The biomechanics of arterial elastin.

Uniaxial mechanical experiments have shown that a neo-Hookean/Gaussian model is suitable to describe the mechanics of arterial elastin networks [Gundiah, N., Ratcliffe, M.B., Pruitt, L.A., 2007. Determination of strain energy function for arterial elastin: Experiments using histology and mechanical tests. J. Biomech. 40, 586-594]. Based on the three-dimensional elastin architecture in arteries, we have proposed an orthotropic material symmetry for arterial elastin consisting of two orthogonally oriented and symmetrically placed families of mechanically equivalent fibers. In this study, we use these results to describe the strain energy function for arterial elastin, with dependence on a reduced subclass of invariants, as W=W(I(1),I(4)). We use previously published equations for this dependence [Humphrey, J.D., Strumpf, R.K., Yin, F.C.P., 1990a. Determination of a constitutive relation for passive myocardium: I. A new functional form. J. Biomech. Eng. 112, 333-339], in combination with a theoretical guided Rivlin-Saunders framework [Rivlin, R.S., Saunders, D.W., 1951. Large elastic deformations of isotropic materials VII. Experiments on the deformation of rubber. Phil. Trans. R. Soc. A 243, 251-288] and biaxial mechanical experiments, to obtain the form of this dependence. Using mechanical equivalence of elastin in the circumferential and longitudinal directions, we add a term in I(6) to W that is similar to the form in I(4). We propose a semi-empirical model for arterial elastin given by W = c(0)(I(1) - 3) + c(1)(I(4) - 1)2 + c(2)(I(6) - 1)2, where c(0), c(1) and c(3) are unknown coefficients. We used the Levenberg-Marquardt algorithm to fit theoretically calculated and experimentally determined stresses from equibiaxial experiments on autoclaved elastin tissues and obtain c(0) = 73.96+/-22.51 kPa, c(1) = 1.18+/-1.79 kPa and c(2) = 0.8+/-1.26 kPa. Thus, the entropic contribution to the strain energy function, represented by c(0), is a dominant feature of elastin mechanics. Because there are no significant differences in the coefficients corresponding to invariants I(4) and I(6), we surmise that there is an equal distribution of fibers in the circumferential and axial directions.

ATR-FTIR as a thickness measurement technique for hydrated polymer-on-polymer coatings.

Hydrated polymer coatings on polymer substrates are common for many biomedical applications, such as tissue engineering constructs, contact lenses, and catheters. The thickness of the coatings can affect the mechanical behavior of the systems and the cellular response, but measuring the coating thickness can be quite challenging using conventional methods. We propose a new method, that is, attenuated total reflection Fourier transform infrared spectroscopy (ATR-FTIR) to determine the relative thickness, combined with atomic force microscopy to calibrate the ATR-FTIR measurements. This technique was successfully employed to determine the hydrated thickness of a series of crosslinked tetraglyme coatings on ultrahigh molecular weight polyethylene substrates intended to reduce wear of acetabular cups in total hip replacements. The hydrated coatings ranged from 30 to 200 nm thick and were accurately measured despite the relatively high root-mean-square (RMS) roughness of the substrates, 20-35 nm (peak-to-peak roughness 55-100 nm). The calibrated ATR-FTIR technique is a promising new method for measuring the thickness of many other polymer-on-polymer and hydrated coatings.

Nanomechanical properties of calcification, fibrous tissue, and hematoma from atherosclerotic plaques.

Clinical events such as heart attack and stroke can be caused by the rupture of atherosclerotic plaques in artery walls. Computational modeling is often used to better understand atherosclerotic disease progression to identify "vulnerable" plaques (i.e., those likely to rupture) and to tailor treatments according to tissue composition. However, because of the heterogeneity of plaque tissue, there are limited data available on the material properties of individual plaque constituents. The goal of this study was to use nanoindentation to measure the mechanical properties of blood clots, fibrous tissue, partially calcified fibrous tissue, and bulk calcifications from human atherosclerotic plaque tissue. Fourier transform infrared (FTIR) spectroscopy was used to quantify the amount of mineral and lipid in each tissue region tested. The results demonstrate that the stiffness of plaque tissue increases with increasing mineral content. In addition, by providing the first experimental data on atherosclerotic calcifications, these data show that some of the estimated modulus values commonly used in computational models greatly underestimate the stiffness of the fully calcified tissue.

Nanoindentation differentiates tissue-scale functional properties of native articular cartilage.

Cartilage mechanical properties are typically tested at the macroscale. To demonstrate the ability of nanoindentation to characterize in situ articular cartilage properties at the tissue scale, we investigated the local structure-property relationships of intact articular cartilage of a normal rabbit metacarpophalangeal joint. We calculated the mechanical parameters of stiffness, S, resistance to penetration, R, and volumetric creep strain, dV/V, from nanoindentation of the articular surface at specific regions of interest. We measured morphological parameters of superficial zone thickness, middle zone thickness, total uncalcified thickness, and cell density from corresponding regions with light and polarized light microscopy. Mechanical parameters were compared to morphological parameters. There were significant positive correlations (r = 0.98, p < 0.05) between superficial zone thickness and both S and R. However, we found no significant correlation between dV/V and the zone sizes. There were moderate, negative correlations between cell density and both S and R, suggesting an effect of cell volume on cartilage behavior at the tissue scale. We opine that the superficial zone plays important role in load support, as evidenced by correlations between zone size and intact cartilage mechanical properties.