Physiologic chemoattractant gradients are shaped by diffusion, advection, binding to an extracellular matrix, and removal by cells. Previous in vitro tools for studying these gradients and the ...cellular migratory response have required cells to be constrained to a 2D substrate or embedded in a gel devoid of fluid flow. Cell migration in fluid flow has been quantified in the absence of chemoattractant gradients and shown to be responsive to them, but there is a need for tools to investigate the synergistic, or antagonistic, effects of gradients and flow. We present a microfluidic chip in which we generated precisely controlled gradients of the chemokine CCL19 under advective-diffusive conditions. Using torque-actuated membranes situated between a gel region and the chip outlet, the resistance of fluid channels adjacent to the gel region could be modified, creating a controllable pressure difference across the gel at a resolution inferior to 10 Pa. Constant supply and removal of chemokine on either side of the chip facilitated the formation of stable gradients at Péclet numbers between −10 and +10 in a collagen type I hydrogel. The resulting interstitial flow was steady within 0.05 μm s−1 for at least 8 h and varied by less than 0.05 μm s−1 along the gel region. This method advances the physiologic relevance of the study of the formation and maintenance of molecular gradients and cell migration, which will improve the understanding of in vivo observations.
All protective and pathogenic immune and inflammatory responses rely heavily on leukocyte migration and localization. Chemokines are secreted chemoattractants that orchestrate the positioning and ...migration of leukocytes through concentration gradients. The mechanisms underlying chemokine gradient establishment and control include physical as well as biological phenomena. Mathematical models offer the potential to both understand this complexity and suggest interventions to modulate immune function. Constructing models that have powerful predictive capability relies on experimental data to estimate model parameters accurately, but even with a reductionist approach most experiments include multiple cell types, competing interdependent processes and considerable uncertainty. Therefore, we propose the use of reduced modeling and experimental frameworks in complement, to minimize the number of parameters to be estimated. We present a Bayesian optimization framework that accounts for advection and diffusion of a chemokine surrogate and the chemokine CCL19, transport processes that are known to contribute to the establishment of spatio-temporal chemokine gradients. Three examples are provided that demonstrate the estimation of the governing parameters as well as the underlying uncertainty. This study demonstrates how a synergistic approach between experimental and computational modeling benefits from the Bayesian approach to provide a robust analysis of chemokine transport. It provides a building block for a larger research effort to gain holistic insight and generate novel and testable hypotheses in chemokine biology and leukocyte trafficking.
There has existed a severe ventilator deficit in much of the world for many years, due in part to the high cost and complexity of traditional ICU ventilators. This was highlighted and exacerbated by ...the emergence of the COVID-19 pandemic, during which the increase in ventilator production rapidly overran the global supply chains for components. In response, we propose a new approach to ventilator design that meets the performance requirements for COVID-19 patients, while using components that minimise interference with the existing ventilator supply chains. The majority of current ventilator designs use proportional valves and flow sensors, which remain in short supply over a year into the pandemic. In the proposed design, the core components are on-off valves. Unlike proportional valves, on-off valves are widely available, but accurate control of ventilation using on-off valves is not straightforward. Our proposed solution combines four on-off valves, a two-litre reservoir, an oxygen sensor and two pressure sensors. Benchtop testing of a prototype was performed with a commercially available flow analyser and test lungs. We investigated the accuracy and precision of the prototype using both compressed gas supplies and a portable oxygen concentrator, and demonstrated the long-term durability over 15 days. The precision and accuracy of ventilation parameters were within the ranges specified in international guidelines in all tests. A numerical model of the system was developed and validated against experimental data. The model was used to determine usable ranges of valve flow coefficients to increase supply chain flexibility. This new design provides the performance necessary for the majority of patients that require ventilation. Applications include COVID-19 as well as pneumonia, influenza, and tuberculosis, which remain major causes of mortality in low and middle income countries. The robustness, energy efficiency, ease of maintenance, price and availability of on-off valves are all advantageous over proportional valves. As a result, the proposed ventilator design will cost significantly less to manufacture and maintain than current market designs and has the potential to increase global ventilator availability.
Intravital microscopy and other direct-imaging techniques have allowed for a characterisation of leukocyte migration that has revolutionised the field of immunology, resulting in an unprecedented ...understanding of the mechanisms of immune response and adaptive immunity. However, there is an assumption within the field that modern imaging techniques permit imaging parameters where the resulting cell track accurately captures a cell's motion. This notion is almost entirely untested, and the relationship between what could be observed at a given scale and the underlying cell behaviour is undefined. Insufficient spatial and temporal resolutions within migration assays can result in misrepresentation of important physiologic processes or cause subtle changes in critical cell behaviour to be missed. In this review, we contextualise how scale can affect the perceived migratory behaviour of cells, summarise the limited approaches to mitigate this effect, and establish the need for a widely implemented framework to account for scale and correct observations of cell motion. We then extend the concept of scale to new approaches that seek to bridge the current "black box" between single-cell behaviour and systemic response.
Abstract
OBJECTIVES
This study aims to characterize the material properties of ascending thoracic aortic aneurysmal tissue, using regional biomechanical assessment of both tensile and dissection ...propagation peel strength.
METHODS
Thirty-four aneurysm specimens (proximal thoracic aorta) were harvested en-bloc from patients undergoing surgery for aneurysm replacement. Specimens were processed into regional samples of similar shapes covering the whole aneurysm isosurface, according to a structured protocol, in both orientations (longitudinal and circumferential). Thickness mapping, uniaxial tensile and peel tests were conducted, enabling calculation of the following parameters: true stress/strain, tangential modulus, tensile strength, peeling force and dissection energy. Two constitutive material models were used (hyperelastic models of Delfino and Ogden) to fit the data. A circumferential strip of tissue was also obtained for computational histology regional quantification of (i) elastin, (ii) collagen and (iii) smooth muscle cells.
RESULTS
The aortic wall was thinner on the outer curve (2.21, standard deviation (SD) 0.4 mm vs inner curve 2.50, SD 0.12 mm). Advanced patient age and higher pulse wave velocity (externally measured) were predictors of increased aortic wall thickness. Tensile strength was higher in the circumferential versus longitudinal direction when analysed according to anatomical regions. Both peel force (35.5, 22 N/m) and dissection energy (88.5, 69 J/m2) were on average lowest at the outer curve of the aneurysm in the longitudinal orientation. Delfino and Ogden model constants varied throughout anatomical regions, with the outer curve being associated a higher ɑ constant (Delfino) and lower µ1 constant (Ogden) (P < 0.05) indicating increased stiffness. Histologically, collagen abundance was significantly related to circumferential and longitudinal strength (P= 0.010), whilst smooth muscle cell count had no relation with any mechanical property (P > 0.05).
CONCLUSIONS
Our results suggest that the outer aortic curve is more prone to dissection propagation and perhaps less prone to rupture than the inner aortic curve. This strengthens the notion of disease heterogeneity in ascending thoracic aortic aneurysms and has implications for the pathogenesis of aortic dissection.
The unmet clinical need in the management of ascending thoracic aortic aneurysms (ATAA) is the lack of an accurate predictive model or biomarker for the risk of acute aortic syndrome 1, 2, Current guidelines rely only on aneurysm size and growth to determine the risk of type A aortic dissection (TAAD) 3.
Display omitted
Biodegradable stents show promise to revolutionize coronary artery disease treatment. Its successful implementation in the global market remains limited due to the constraints of ...current generation biodegradable materials. Cold gas dynamic spraying (CGDS) has been proposed as a manufacturing approach to fabricate a metallic biodegradable amalgamate for stent application. Iron and 316L stainless steel powders are combined in a 4:1 ratio to create a novel biomaterial through cold spray. Cold spray processing however, produces a coating in a work hardened state, with limited ductility, which is a critical mechanical property in stent design. To this end, the influence of annealing temperature on the mechanical and corrosion performances of the proposed Fe-316L amalgamate is investigated. It was found that annealing at 1300 °C yielded a complex material microstructure, with an ultimate tensile strength of approximately 280 MPa and ductility of 23%. The static corrosion rate determined at this annealing temperature was equal to 0.22 mg cm−2 day−1, with multiple corrosion species identified within the degradation layers. Precipitates were observed throughout the microstructure, which appeared to accelerate the overall corrosion behaviour. It was shown that cold-sprayed Fe-316L has significant potential to be implemented in a clinical setting.
Biodegradable stents have potential to significantly improve treatment of coronary artery disease by decreasing or potentially eliminating late-term complications, including stent fracture and in-stent restenosis. Current generation polymer biodegradable stents have led to poorer patient outcomes in comparison to drug-eluting stents, however, and it is evident that metallic biomaterials are required, which have increased strength. To this end, a novel iron and stainless steel 316L biomaterial is proposed, fabricated through cold-gas dynamic spraying. This study analyses the effect of annealing on the Fe-316L biomaterial through corrosion, mechanical, and microstructural investigations. The quantitative data presented in this work suggests that Fe-316L, in its annealed condition, has the mechanical and corrosion properties necessary for biodegradable stent application.
Swelling of lymph nodes (LNs) is commonly observed during the adaptive immune response, yet the impact on T cell (TC) trafficking and subsequent immune response is not well known. To better ...understand the effect of macro-scale alterations, we developed an agent-based model of the LN paracortex, describing the TC proliferative response to antigen-presenting dendritic cells alongside inflammation-driven and swelling-induced changes in TC recruitment and egress, while also incorporating regulation of the expression of egress-modulating TC receptor sphingosine-1-phosphate receptor-1. Analysis of the effector TC response under varying swelling conditions showed that swelling consistently aided TC activation. However, subsequent effector CD8
TC production was reduced in scenarios where swelling occurred too early in the TC proliferative phase or when TC cognate frequency was low due to increased opportunity for TC exit. Temporarily extending retention of newly differentiated effector TCs, mediated by sphingosine-1-phosphate receptor-1 expression, mitigated any negative effects of swelling by allowing facilitation of activation to outweigh increased access to exit areas. These results suggest that targeting temporary effector TC retention and egress associated with swelling offers new ways to modulate effector TC responses in, for example, immuno-suppressed patients and to optimize of vaccine design.
Strain measurement during tissue deformation is crucial to elucidate relationships between mechanical loading and functional changes in biological tissues. When combined with specified loading ...conditions, assessment of strain fields can be used to craft models that accurately represent the mechanical behavior of soft tissue. Inhomogeneities in strain fields may be indicative of normal or pathological inhomogeneities in mechanical properties. In this study, we present the validation of a modified Demons registration algorithm for non-contact, marker-less strain measurement of tissue undergoing uniaxial loading. We validate the algorithm on a synthetic dataset composed of artificial deformation fields applied to a speckle image, as well as images of aortic sections of varying perceptual quality. Initial results indicate that Demons outperforms recent Optical Flow and Digital Image Correlation methods in terms of accuracy and robustness to low image quality, with similar runtimes. Demons achieves at least 8% lower maximal deviation from ground truth on 50% biaxial and shear strain applied to aortic images. To illustrate utility, we quantified strain fields of multiple human aortic specimens undergoing uniaxial tensile testing, noting the formation of strain concentrations in areas of rupture. The modified Demons algorithm captured a large range of strains (up to 50%) and provided spatially resolved strain fields that could be useful in the assessment of soft tissue pathologies.
The application of biodegradable materials to stent design has the potential to transform coronary artery disease treatment. It is critical that biodegradable stents have sustained strength during ...degradation and vessel healing to prevent re-occlusion. Proper assessment of the impact of corrosion on the mechanical behaviour of potential biomaterials is important. Investigations within literature frequently implement simplified testing conditions to understand this behaviour and fail to consider size effects associated with strut thickness, or the increase in corrosion due to blood flow, both of which can impact material properties. A protocol was developed that utilizes micro-scale specimens, in conjunction with dynamic degradation, to assess the effect of corrosion on the mechanical properties of a novel Fe-316L material. Dynamic degradation led to increased specimen corrosion, resulting in a greater reduction in strength after 48 h of degradation in comparison to samples statically corroded. It was found that thicker micro-tensile samples (h > 200 μm) had a greater loss of strength in comparison to its thinner counterpart (h < 200 μm), due to increased corrosion of the thicker samples (203 MPa versus 260 MPa after 48 h, p = 0.0017). This investigation emphasizes the necessity of implementing physiologically relevant testing conditions, including dynamic corrosion and stent strut thickness, when evaluating potential biomaterials for biodegradable stent application.
Display omitted
•Development of micro-tensile protocol in conjunction with dynamic corrosion tests.•Effect of corrosion on mechanical properties of Fe-316L influenced by specimen size.•Dynamic degradation had greater impact than static corrosion environment.•Thicker micro-samples had greater reduction in strength than thinner micro-samples.
According to the World Health Organization, coronary artery disease is the leading cause of death in the developed world. Percutaneous coronary intervention with stent insertion is the most widely ...used therapy to treat coronary artery disease. The trend in stent design has seen a shift from permanent metallic stents to the implementation of biodegradable materials. The current generation of polymer biodegradable stents approved for clinical use, however, have faced significant challenges due to their poor mechanical properties and resulting thicker struts, leading to comparatively poor patient outcomes. Cold gas dynamic spraying was proposed as an alternate fabrication method to manufacture a novel material for biodegradable stent application. Cold spray is advantageous as it produces coatings with a refined microstructure, leading to increased strength and fatigue resistance. Iron and stainless steel 316L powders were mixed in a 4:1 ratio to produce an intermixed material, where the corrosion rate was controlled by the relative weight percentages of the powders and accelerated due to the galvanic couple between Fe and 316L particles. However, due to the deposition process of cold-sprayed materials, it is in a severely work hardened state, with limited ductility, which is a critical material property for stent application. Therefore, it was the primary objective of this thesis to develop a manufacturing procedure that would improve the mechanical properties of cold-sprayed Fe-316L for stent application. It was determined that annealing at 1300 °C for two hours resulted in an tensile strength of approximately 300 MPa and a ductility of 23%, which was a significant increase in comparison to as-sprayed Fe-316L (0.05%). Microstructural observation revealed a phase transformation of 316L particles from austenite to ferrite after annealing, with substantial atomic diffusion. Corrosion tests indicated that the degradation rate had decreased after heat treatment in comparison to as-sprayed Fe-316L, due to an improvement of particle-particle bonds and due to the loss of the galvanic couple between Fe and 316L particles. The corrosion rate of the Fe-316L annealed at 1300 °C for 2 hours was 0.22 mg/(cm2 day). A series of investigations were also conducted to assess the effect of corrosion on the mechanical properties of annealed Fe-316L, using macro- and micro-sized samples. A study utilizing bulk specimens indicated that static corrosion had a minimal effect on the mechanical properties of Fe-316L, with only a statistically significant difference in tensile strength after 7 days of static corrosion testing. In comparison, a more significant decrease in UTS was observed for micro-sized samples subjected to dynamic corrosion. It was observed that thicker samples had increased corrosion and reduction in strength than thinner micro specimens. It was demonstrated in this thesis that Fe-316L, fabricated through cold spray, had the material properties necessary for biodegradable stent application. Overall, the mechanical properties of annealed Fe-316L were superior to that of current generation polymer stents. While the galvanic couple between iron and 316L particles was lost due to phase transformation and significant atomic diffusion, there was an indication of microgalvanic corrosion with observed precipitates and the surrounding matrix, which requires further study to understand its effect. The preliminary micro-tensile tests suggested that there was not a significant size effect between the bulk and micro-sized samples, indicative that a reduced stent strut thickness is feasible. A manufacturing protocol was developed that allowed fabrication of stents with a strut thickness of 105 µm. In conclusion, it has been shown that an Fe-316L cold-sprayed stent has significant potential to be implemented in a clinical setting, and future investigations should focus on pre-clinical and clinical trials to bring the stent to the global market.
PDF
