Mechanical testing of glutaraldehyde cross-linked mitral valves. Part one: In vitro mechanical behaviour.

The aim of this study was to perform an initial assessment, in vitro, of the feasibility of using a glutaraldehyde cross-linked porcine mitral valve to retain acute functionality, focusing on assessing mitral regurgitation. Six porcine hearts were tested using an in vitro simulator. Testing was repeated following cross-linking of mitral valves; where cross-linking was achieved by placing them in a glutaraldehyde solution. The simulator enabled systolic pressure on the ventricular side of the valve to be mimicked. Following testing, mitral valve leaflets underwent Scanning Electron Microscopy of the ventricular surface of both the anterior and posterior leaflets (1 cm2 samples). The peak pressure withstood by cross-linked valves was significantly lower than for untreated valves (108 mmHg cf. 128 mmHg for untreated valves; p < 0.05). The peak pressure was typically reached 0.5 s later than for the untreated valve. While both cross-linked and untreated valves exhibited endothelium denudation, the unfixed valve had less endothelial loss. Glutaraldehyde cross-linking of porcine mitral valves may be of potential value in assessing improved bioprosthetic mitral valve replacements. However, a more immobile valve exhibiting endothelial denudation (i.e. sclerosis) was a possible concerns identified following in vitro acute assessment.

Mechanical testing of glutaraldehyde cross-linked mitral valves. Part two: Elastic and viscoelastic properties of chordae tendineae.

The aim of this study was to assess whether the mechanical properties of mitral valve chordae tendineae are sensitive to being cross-linked under load. A total 64 chordae were extracted from eight porcine hearts. Two chordae (posterior basal) from each heart were subjected to uniaxial ramp testing and six chordae (two strut, two anterior basal and two posterior basal) were subjected to dynamic mechanical analysis over frequencies between 0.5 and 10 Hz. Chordae were either cross-linked in tension or cross-linked in the absence of loading. Chordae cross-linked under load transitioned from high to low extension at a lower strain than cross-linked unloaded chordae (0.07 cf. 0.22), with greater pre-transitional (30.8 MPa cf. 5.78 MPa) and post-transitional (139 MPa cf. 74.1 MPa) moduli. The mean storage modulus of anterior strut chordae ranged from 48 to 54 MPa for cross-linked unloaded chordae, as compared to 53-61 MPa cross-linked loaded chordae. The mean loss modulus of anterior strut chordae ranged from 2.3 to 2.9 MPa for cross-linked unloaded chordae, as compared to 3.8-4.8 MPa cross-linked loaded chordae. The elastic and viscoelastic properties of chordae following glutaraldehyde cross-linking are dependent on the inclusion/exclusion of loading during the cross-linking process; with loading increasing the magnitude of the material properties measured.

Multiscale three-dimensional surface reconstruction and surface roughness of porcine left anterior descending coronary arteries.

The aim of this study was to investigate the multiscale surface roughness characteristics of coronary arteries, to aid in the development of novel biomaterials and bioinspired medical devices. Porcine left anterior descending coronary arteries were dissected ex vivo, and specimens were chemically fixed and dehydrated for testing. Surface roughness was calculated from three-dimensional reconstructed surface images obtained by optical, scanning electron and atomic force microscopy, ranging in magnification from 10× to 5500×. Circumferential surface roughness decreased with magnification, and microscopy type was found to influence surface roughness values. Longitudinal surface roughness was not affected by magnification or microscopy types within the parameters of this study. This study found that coronary arteries exhibit multiscale characteristics. It also highlights the importance of ensuring consistent microscopy parameters to provide comparable surface roughness values.

The Effect of Mechanical Overloading on Surface Roughness of the Coronary Arteries.

BACKGROUND: Surface roughness can be used to identify disease within biological tissues. Quantifying surface roughness in the coronary arteries aids in developing treatments for coronary heart disease. This study investigates the effect of extreme physiological loading on surface roughness, for example, due to a rupture of an artery. METHODS: The porcine left anterior descending (LAD) coronary arteries were dissected ex vivo. Mechanical overloading was applied to the arteries in the longitudinal direction to simulate extreme physiological loading. Surface roughness was calculated from three-dimensional reconstructed images. Surface roughness was measured before and after damage and after chemical processing to dehydrate tissue specimens. RESULTS: Control specimens confirmed that dehydration alone results in an increase of surface roughness in the circumferential direction only. No variation was noted between the hydrated healthy and damaged specimens, in both the longitudinal (0.91 ± 0.26 and 1.05 ± 0.25 µm) and circumferential (1.46 ± 0.38 and 1.47 ± 0.39 µm) directions. After dehydration, an increase in surface roughness was noted for damaged specimens in both the longitudinal (1.28 ± 0.33 µm) and circumferential (1.95 ± 0.56 µm) directions. CONCLUSIONS: Mechanical overloading applied in the longitudinal direction did not significantly affect surface roughness. However, when combined with chemical processing, a significant increase in surface roughness was noted in both the circumferential and longitudinal directions. Mechanical overloading causes damage to the internal constituents of the arteries, which is significantly noticeable after dehydration of tissue.

Objective Uniaxial Identification of Transition Points in Non-Linear Materials: Sample Application to Porcine Coronary Arteries and the Dependency of Their Pre- and Post-Transitional Moduli with Position.

PURPOSE: This study aimed to develop an objective method for the elastic characterisation of pre- and post-transitional moduli of left anterior descending (LAD) porcine coronary arteries. METHODS: Eight coronary arteries were divided into proximal, middle and distal test specimens. Specimens underwent uniaxial extension up to 3 mm. Force-displacement measurements were used to determine the induced true stress and stretch for each specimen. A local maximum of the stretch-true stress data was used to identify a transition point. Pre- and post-transitional moduli were calculated up to and from this point, respectively. RESULTS: The mean pre-transitional moduli for all specimens was 0.76 MPa, as compared to 4.86 MPa for the post-transitional moduli. However, proximal post-transitional moduli were significantly greater than that of middle and distal test specimens (p < 0.05). CONCLUSION: Post-transitional uniaxial properties of the LAD are dependent on location along the artery. Further, it is feasible to objectively identify a transition point between pre- and post-transitional moduli.

The design of additively manufactured lattices to increase the functionality of medical implants.

The rise of antibiotic resistant bacterial species is driving the requirement for medical devices that minimise infection risks. Antimicrobial functionality may be achieved by modifying the implant design to incorporate a reservoir that locally releases a therapeutic. For this approach to be successful it is critical that mechanical functionality of the implant is maintained. This study explores the opportunity to exploit the design flexibilities possible using additive manufacturing to develop porous lattices that maximise the volume available for drug loading while maintaining load-bearing capacity of a hip implant. Eight unit cell types were initially investigated and a volume fraction of 30% was identified as the lowest level at which all lattices met the design criteria in ISO 13314. Finite element analysis (FEA) identified three lattice types that exhibited significantly lower displacement (10-fold) compared with other designs; Schwartz primitive, Schwartz primitive pinched and cylinder grid. These lattices were additively manufactured in Ti-6Al-4V using selective laser melting. Each design exceeded the minimum strength requirements for orthopaedic hip implants according to ISO 7206-4. The Schwartz primitive (Pinched) lattice geometry, with 10% volume fill and a cubic unit cell period of 10, allowed the greatest void volume of all lattice designs whilst meeting the fatigue requirements for use in an orthopaedic implant (ISO 7206-4). This paper demonstrates an example of how additive manufacture may be exploited to add additional functionality to medical implants.

Hyperspectral chemical imaging reveals spatially varied degradation of polycarbonate urethane (PCU) biomaterials.

Hyperspectral chemical imaging (HCI) is an emerging technique which combines spectroscopy with imaging. Unlike traditional point spectroscopy, which is used in the majority of polymer biomaterial degradation studies, HCI enables the acquisition of spatially localised spectra across the surface of a material in an objective manner. Here, we demonstrate that attenuated total reflectance Fourier transform infra-red (ATR-FTIR) HCI reveals spatial variation in the degradation of implantable polycarbonate urethane (PCU) biomaterials. It is also shown that HCI can detect possible defects in biomaterial formulation or specimen production; these spatially resolved images reveal regional or scattered spatial heterogeneity. Further, we demonstrate a map sampling method, which can be used in time-sensitive scenarios, allowing for the investigation of degradation across a larger component or component area. Unlike imaging, mapping does not produce a contiguous image, yet grants an insight into the spatial heterogeneity of the biomaterial across a larger area. These novel applications of HCI demonstrate its ability to assist in the detection of defective manufacturing components and lead to a deeper understanding of how a biomaterial's chemical structure changes due to implantation. STATEMENT OF SIGNIFICANCE: The human body is an aggressive environment for implantable devices and their biomaterial components. Polycarbonate urethane (PCU) biomaterials in particular were investigated in this study. Traditionally one or a few points on the PCU surface are analysed using ATR-FTIR spectroscopy. However the selection of acquisition points is susceptible to operator bias and critical information can be lost. This study utilises hyperspectral chemical imaging (HCI) to demonstrate that the degradation of a biomaterial varies spatially. Further, HCI revealed spatial variations of biomaterials that were not subjected to oxidative degradation leading to the possibility of HCI being used in the assessment of biomaterial formulation and/or component production.

Towards Additive Manufacture of Functional, Spline-Based Morphometric Models of Healthy and Diseased Coronary Arteries: In Vitro Proof-of-Concept Using a Porcine Template.

The aim of this study is to assess the additive manufacture of morphometric models of healthy and diseased coronary arteries. Using a dissected porcine coronary artery, a model was developed with the use of computer aided engineering, with splines used to design arteries in health and disease. The model was altered to demonstrate four cases of stenosis displaying varying severity, based on published morphometric data available. Both an Objet Eden 250 printer and a Solidscape 3Z Pro printer were used in this analysis. A wax printed model was set into a flexible thermoplastic and was valuable for experimental testing with helical flow patterns observed in healthy models, dominating the distal LAD (left anterior descending) and left circumflex arteries. Recirculation zones were detected in all models, but were visibly larger in the stenosed cases. Resin models provide useful analytical tools for understanding the spatial relationships of blood vessels, and could be applied to preoperative planning techniques, but were not suitable for physical testing. In conclusion, it is feasible to develop blood vessel models enabling experimental work; further, through additive manufacture of bio-compatible materials, there is the possibility of manufacturing customized replacement arteries.

Effects of freezing, fixation and dehydration on surface roughness properties of porcine left anterior descending coronary arteries.

BACKGROUND: To allow measurements of surface roughness to be made of coronary arteries using various imaging techniques, chemical processing, such as fixation and dehydration, is commonly used. Standard protocols suggest storing fresh biological tissue at -40°C. The aim of this study was to quantify the changes caused by freezing and chemical processing to the surface roughness measurements of coronary arteries, and to determine whether correction factors are needed for surface roughness measurements of coronary arteries following chemical processes typically used before imaging these arteries. METHODS: Porcine left anterior descending coronary arteries were dissected ex vivo. Surface roughness was then calculated following three-dimensional reconstruction of surface images obtained using an optical microscope. Surface roughness was measured before and after a freeze cycle to assess changes during freezing, after chemical fixation, and again after dehydration, to determine changes during these steps of chemical processing. RESULTS: No significant difference was caused due to the freeze cycle (p>0.05). There was no significant difference in the longitudinally measured surface roughness (RaL=0.99±0.39µm; p>0.05) of coronary arteries following fixation and dehydration either. However, the circumferentially measured surface roughness increased significantly following a combined method of processing (RaC=1.36±0.40, compared 1.98±0.27µm, respectively; p<0.05). A correction factor can compensate for the change RaCß=RaC1+0.46in RaC due to processing of tissue, Where RaCß, the corrected RaC, had a mean of 1.31±0.21µm. CONCLUSIONS: Independently, freezing, fixation and dehydration do not alter the surface roughness of coronary arteries. Combined, however, fixation and dehydration significantly increase the circumferential, but not longitudinal, surface roughness of coronary arteries.

Dynamic Viscoelasticity and Surface Properties of Porcine Left Anterior Descending Coronary Arteries.

The aim of this study was, for the first time, to measure and compare quantitatively the viscoelastic properties and surface roughness of coronary arteries. Porcine left anterior descending coronary arteries were dissected ex vivo. Viscoelastic properties were measured longitudinally using dynamic mechanical analysis, for a range of frequencies from 0.5 to 10 Hz. Surface roughness was calculated following three-dimensional reconstructed of surface images obtained using an optical microscope. Storage modulus ranged from 14.47 to 25.82 MPa, and was found to be frequency-dependent, decreasing as the frequency increased. Storage was greater than the loss modulus, with the latter found to be frequency-independent with a mean value of 2.10 ± 0.33 MPa. The circumferential surface roughness was significantly greater (p < 0.05) than the longitudinal surface roughness, ranging from 0.73 to 2.83 and 0.35 to 0.92 µm, respectively. However, if surface roughness values were corrected for shrinkage during processing, circumferential and longitudinal surface roughness were not significantly different (1.04 ± 0.47, 0.89 ± 0.27 µm, respectively; p > 0.05). No correlation was found between the viscoelastic properties and surface roughness. It is feasible to quantitatively measure the viscoelastic properties of coronary arteries and the roughness of their endothelial surface.