Advanced Material Catheter (AMCath), a minimally invasive endocardial catheter for the delivery of fast-gelling covalently cross-linked hyaluronic acid hydrogels.

Injectable hydrogels that aim to mechanically stabilise the weakened left ventricle wall to restore cardiac function or to deliver stem cells in cardiac regenerative therapy have shown promising data. However, the clinical translation of hydrogel-based therapies has been limited due to difficulties injecting them through catheters. We have engineered a novel catheter, Advanced Materials Catheter (AMCath), that overcomes translational hurdles associated with delivering fast-gelling covalently cross-linked hyaluronic acid hydrogels to the myocardium. We developed an experimental technique to measure the force required to inject such hydrogels and determined the mechanical/viscoelastic properties of the resulting hydrogels. The preliminary in vivo feasibility of delivering fast-gelling hydrogels through AMCath was demonstrated by accessing the porcine left ventricle and showing that the hydrogel was retained in the myocardium post-injection (three 200 µL injections delivered, 192, 204 and 183 µL measured). However, the mechanical properties of the hydrogels were reduced by passage through AMCath (≤20.62% reduction). We have also shown AMCath can be used to deliver cardiopoietic adipose-derived stem cell-loaded hydrogels without compromising the viability (80% viability) of the cells in vitro. Therefore, we show that hydrogel/catheter compatibility issues can be overcome as we have demonstrated the minimally invasive delivery of a fast-gelling covalently cross-linked hydrogel to the beating myocardium.

Freeze-Drying as a Novel Biofabrication Method for Achieving a Controlled Microarchitecture within Large, Complex Natural Biomaterial Scaffolds.

The biofabrication of large natural biomaterial scaffolds into complex 3D shapes which have a controlled microarchitecture remains a major challenge. Freeze-drying (or lyophilization) is a technique used to generate scaffolds in planar 3D geometries. Here we report the development of a new biofabrication process to form a collagen-based scaffold into a large, complex geometry which has a large height to width ratio, and a controlled porous microarchitecture. This biofabrication process is validated through the successful development of a heart valve shaped scaffold, fabricated from a collagen-glycosaminoglycan co-polymer. Notably, despite the significant challenges in using freeze-drying to create such a structure, the resultant scaffold has a uniform, homogenous pore architecture throughout. This is achieved through optimization of the freeze-drying mold and the freezing parameters. We believe this to be the first demonstration of using freeze-drying to create a large, complex scaffold geometry with a controlled, porous architecture for natural biomaterials. This study validates the potential of using freeze-drying for development of organ-specific scaffold geometries for tissue engineering applications, which up until now might not have been considered feasible.

The development and mechanical characterisation of a novel reinforced venous conduit that mimics the mechanical properties of an arterial wall.

Venous grafts have been used to bypass stenotic arteries for many decades. However, this "gold standard" treatment is far from optimal, with long-term vein graft patency rates reported to be as low as 50% at >15 years. These results could be a result of the structural and functional differences of veins compared to arteries. In this study we developed a new protocol for manufacturing reinforced fresh veins with a decellularized porcine arterial scaffold. This novel method was designed to be replicated easily in a surgical setting, and manufactured reinforced constructs were robust and easier to handle than the veins alone. Furthermore, we demonstrate that these Reinforced Venous-Arterial Conduits have comparable mechanical properties to native arteries, in terms of ultimate tensile strength (UTS) (2.36 vs. 2.24MPa) and collagen dominant phase (11.04 vs. 12.26MPa). Therefore, the Reinforced Venous-Arterial Conduit combines the benefits of using the current gold standard homogenous venous grafts composed of a confluent endothelial surface, with an "off-the-shelf" decellularized artery to improve the mechanical properties to closely mimic those of native arteries, while maintaining the self-repairing characteristics of native tissue. In conclusion in this study we have produced a construct and a new technique that combines the mechanical properties of both a natural vein and a decellularized artery to produce a reinforced venous graft that closely mimics the mechanical response of an arterial segment.

The suture pullout characteristics of human and porcine linea alba.

There is a substantial prevalence of post-operative incisional hernia for both laparoscopic and laparotomy procedures, but there have been few attempts at quantifying abdominal wound closure methodology in the literature. One method to ascertain a more robust method of wound closure is the identification of the influence of suture placement parameters on suture pullout force. Current surgical practice involves a recommended bite depth and bite separation of 10mm, but the evidence base for this is not clear. In this paper, the suture pullout characteristics of both porcine and human linea alba were investigated to ascertain a suture placement protocol for surgical wound closure. Uniaxial suture pullout force testing on fresh frozen porcine and human linea alba samples was performed using standard materials testing machines. The influence of the number of suture loops, the bite depth and the bite separation of the sutures and the orientation of the sutures with respect to the principal fibre direction in the linea alba were assessed. Results showed a clearly identifiable relationship between pullout force of the suture, bite separation and bite depth, with low suture separation and high suture depth as optimal parameters for increasing pullout force. Resistance to pullout could be improved by as much as 290% when optimizing test conditions. Both human and porcine tissue were observed to exhibit very similar pullout force characteristics, corroborating the use of a porcine model for investigations into wound closure methodology. Orientation of suture application was also found to significantly affect the magnitude of suture pullout, with suturing applied longitudinally across a transverse defect resulting in higher pullout forces for small suture bite separations. Although further assessment in an environment more representative of in vivo conditions is required, these findings indicate that increasing the bite depth and reducing the bite separation with respect to the current surgical recommendations may reduce the risk of post-operative incisional hernia.

Uniaxial and biaxial tensile stress-stretch response of human linea alba.

There are few studies on the stress-stretch behaviour of human linea alba, yet understanding the mechanics of this tissue is important for developing better methods of abdominal wound closure. Published data focuses mainly on porcine linea alba and for human tissue there are conflicting results and no bi-axial data available. This variability is likely due to challenges with the physical dimensions of the tissue and differences in experimental methodology. This study focussed on the tensile mechanical characterisation of the human linea alba using uniaxial and equi-load biaxial testing performed using image-based strain measurement methods. Thirteen freshly frozen human cadaveric abdominal walls were obtained and used to prepare 7 samples in both the transverse and longitudinal directions for uniaxial testing, and 13 square samples for bi-axial testing. The results showed significant anisotropy and for the equi-load biaxial tests the deformation was heavily biased in the longitudinal direction. In comparison with similar tests on porcine tissue from a previous study, it was found that the response of porcine linea alba to uniaxial loading is similar to that of human linea alba, with no statistically significant differences observed. Under biaxial loading human and porcine linea showed no statistical significance in the difference between their means in the transverse direction. However, a significant difference was observed in the longitudinal direction, and further study of the respective tissue structures is needed to better understand this result. These results provide the first data on the biaxial tensile properties of human linea alba and can aid in an improved assessment of wound closure mechanics.

Uniaxial and biaxial mechanical properties of porcine linea alba.

Incisional hernia is a severe complication post-laparoscopic/laparotomy surgery that is commonly associated with the linea alba. However, the few studies on the mechanical properties of the linea alba in the literature appear contradictory, possible due to challenges with the physical dimensions of samples and variations in protocol. This study focuses on the tensile mechanical characterisation of the porcine linea alba, as determined by uniaxial and equi-load biaxial testing using image-based strain measurement methods. Results show that the linea alba demonstrated a non-linear elastic, anisotropic behaviour which is often observed in biological soft tissues. The transverse direction (parallel to fibres) was found to be approximately eight times stiffer than the longitudinal (cross-fibre) direction under both uniaxial and equi-load biaxial loading. The equi-load biaxial tensile tests revealed that contraction could occur in the transverse direction despite increasing load, probably due to the anisotropy of the tissue. Optical surface marker tracking and digital image correlation methods were found to greatly improve the accuracy of stretch measurement, resulting in a 75% change in the apparent stiffness compared to using strain derived from machine cross-head displacement. Additionally, a finite element model of the experiments using a combination of an Ogden and fibre exponential power law model for the linea alba was implemented to quantify the effect of clamping and tissue dimensions (which are suboptimal for tensile testing) on the results. The preliminary model results were used to apply a correction factor to the uniaxial experimental data prior to inverse optimisation to derive best fit material parameters for the fibre reinforced Ogden model. Application of the model to the equi-load biaxial case showed some differences compared to the experimental data, suggesting a more complex anisotropic model may be necessary to capture biaxial behaviour. These results provide an improved assessment of the mechanical properties of the porcine linea alba for wound closure and other studies.

Assessing the microstructural response to applied deformation in porcine passive skeletal muscle.

The passive micro-structural mechanical response of muscle tissue is important for numerous medical applications. However, the recently observed tension/compression asymmetry in porcine muscle remains poorly explained. In particular there remains a lack of understanding of how external tension or compression applied in the fibre or cross-fibre direction translates internally to deformation of muscle fibres and the extra-cellular matrix. Accordingly, fresh porcine skeletal muscle tissue was harvested, deformed by 30% in uniaxial tension or compression in both the fibre and cross-fibre directions and prepared for optical microscope, polarised light microscope and SEM analysis. The average deformed specimen results were compared to the average control results in each case. For compressive or tensile stretch applied in the muscle fibre direction the average measured muscle fibre cross-sectional area changes are in close correspondence with predictions based on global Poisson's ratio measurements and these deformation modes did not cause shape changes in the muscle fibre cross-sections. However, muscle tissue reacted to the applied cross-fibre direction deformations as follows: compression flattened muscle fibre cross-sections, aligning them perpendicular to the direction of the applied deformation while tensile deformations stretched the cross-sections of muscle fibres, aligning them parallel to the direction of applied deformation. No evidence of structural reorganisation of endomysium collagen fibres in response to applied stretch was observed. The observed micro-structural responses do not appear to be influenced by the surrounding endomysium, but appear to be significantly influenced by proximity to the perimysium network. It is hypothesised that the perimysium and its interaction with the surrounding muscle fibres is therefore likely to be predominantly responsible for the tension/compression asymmetry observed in macroscopic tests of passive skeletal muscle stress strain behaviour.

Effects of creatine on isometric bench-press performance in resistance-trained humans.

PURPOSE: The purpose of this study was to investigate the effects of creatine (Cr) supplementation on force generation during an isometric bench-press in resistance-trained men. METHODS: 32 resistance-trained men were matched for peak isometric force and assigned in double-blind fashion to either a Cr or placebo group. Subjects performed an isometric bench-press test involving five maximal isometric contractions before and after 5 d of Cr (20 g.d-1 Cr + 180 g.d-1 dextrose) or placebo (200 g.d-1 dextrose). Body composition was measured before and after supplementation. Subjects completed 24-h urine collections throughout the study period; these were subsequently analyzed to provide total Cr and creatinine excretion. RESULTS: The amount of Cr retained over the supplementation period was 45 +/- 18 g (mean +/- SD), with an estimated intramuscular Cr storage of 43 (13-61) mmol x kg(-1) x dry weight muscle (median [range]). Four subjects in the Cr group were classified as "nonresponders" (< or =21 mmol x kg(-1) x dry weight muscle increase following Cr supplementation) and the remaining 17 subjects were classed as "responders" (> or =32 mmol x kg(-1) x dry weight muscle). For the Cr group, peak force and total force pre- or post-supplementation were not different from placebo. However, when the analysis was confined to the responders, both the change in peak force [Repetition 2: 59(81) N vs -26(85) N; Repetition 3: 45(59) N vs -26(64) N) and the change in total force (Repetition 1: 1471(1274) N vs 209(1517) N; Repetition 2: 1575(1254) N vs 196(1413) N; Repetition 3: 1278(1245) N vs -3(1118) N; Repetition 4: 918(935) N vs -83(1095) N] post-supplementation were significantly greater compared with the placebo group (P < 0.01). For the Cr group, estimated Cr uptake was inversely correlated with training status (r = -0.68, N = 21, P = 0.001). Cr significantly increased body weight (84.1 +/- 8.6 kg pre- vs 85.3 +/- 8.3 kg post-supplementation) and fat-free mass (71.8 +/- 6.0 kg pre- vs 72.6 +/- 6.0 kg post-supplementation), with the magnitude of increase being significantly greater in the responder group than in the placebo group. CONCLUSION: Five days of Cr supplementation increased body weight and fat-free body mass in resistance-trained men who were classified as responders. Peak force and total force during a repeated maximal isometric bench-press test were also significantly greater in the responders compared to the placebo group.