Three dimensional porous scaffolds derived from collagen, elastin and fibrin proteins orchestrate adipose tissue regeneration.

Current gold standard to treat soft tissue injuries caused by trauma and pathological condition are autografts and off the shelf fillers, but they have inherent weaknesses like donor site morbidity, immuno-compatibility and graft failure. To overcome these limitations, tissue-engineered polymers are seeded with stem cells to improve the potential to restore tissue function. However, their interaction with native tissue is poorly understood so far. To study these interactions and improve outcomes, we have fabricated scaffolds from natural polymers (collagen, fibrin and elastin) by custom-designed processes and their material properties such as surface morphology, swelling, wettability and chemical cross-linking ability were characterised. By using 3D scaffolds, we comprehensive assessed survival, proliferation and phenotype of adipose-derived stem cells in vitro. In vivo, scaffolds were seeded with adipose-derived stem cells and implanted in a rodent model, with X-ray microtomography, histology and immunohistochemistry as read-outs. Collagen-based materials showed higher cell adhesion and proliferation in vitro as well as higher adipogenic properties in vivo. In contrast, fibrin demonstrated poor cellular and adipogenesis properties but higher angiogenesis. Elastin formed the most porous scaffold, with cells displaying a non-aggregated morphology in vitro while in vivo elastin was the most degraded scaffold. These findings of how polymers present in the natural polymers mimicking ECM and seeded with stem cells affect adipogenesis in vitro and in vivo can open avenues to design 3D grafts for soft tissue repair.

A Synergistic Relationship between Polycaprolactone and Natural Polymers Enhances the Physical Properties and Biological Activity of Scaffolds.

Biomaterials for tissue engineering include natural and synthetic polymers, but their clinical application is still limited due to various disadvantages associated with the use of these polymers. This uncertainty of the polymeric approach in tissue engineering launches an opportunity to address a key question: can we eliminate the disadvantages of both natural and synthetic polymers by combining them to form a synergistic relationship? To answer this question, we fabricated scaffolds from elastin, collagen, fibrin, and electrospun polycaprolactone (PCL) with different ratios. The material characterization of these scaffolds investigated degradation, water contact angle, angiogenesis by an ex ovo chorion allantoic membrane (CAM) assay, and mechanical and structural properties. Biological activity and specific differentiation pathways (MSC, adipogenic, osteogenic, myogenic, and chondrogenic) were studied by using human adipose-derived stem cells. Results indicated that all composite polymers degraded at a different rate, thus affecting their mechanical integrity. Cell-based assays demonstrated continual proliferative and viable properties of the cells on all seeded scaffolds with the particular initiation of a differentiation pathway among which the PCL/collagen/fibrin composite was the most angiogenic material with maximum vasculature. We were able to tailor the physical and biological properties of PCL-based composites to form a synergistic relationship for various tissue regeneration applications.

A Durable Porcine Pericardial Surgical Bioprosthetic Heart Valve: a Proof of Concept.

Bioprosthetic leaflets made from animal tissues are used in the majority of surgical and transcatheter cardiac valve replacements. This study develops a new surgical bioprosthesis, using porcine pericardial leaflets. Porcine pericardium was obtained from genetically engineered pigs with a mutation in the GGTA-1 gene (GTKO) and fixed in 0.6% glutaraldehyde, and used to develop a new surgical valve design. The valves underwent in vitro hydrodynamic test in a pulse duplicator and high-cycled accelerated wear testing and were evaluated for acute haemodynamics and thrombogenicity in a juvenile sheep implant study for 48 h. The porcine surgical pericardial heart valves (pSPHVs) exhibited excellent hydrodynamics and reached 200 million cycles of in vitro durability, with no observable damage. Juvenile sheep implants demonstrated normal valve function with no acute thrombogenic response for either material. The pSPHV incorporates a minimalistic construction method using a tissue-to-tissue design to cover the stent. This new design is a proof of concept alternative to the use of bovine pericardium and synthetic fabric in surgical bioprosthetic heart valves.

Anatomically realistic ultrasound phantoms using gel wax with 3D printed moulds.

Here we describe methods for creating tissue-mimicking ultrasound phantoms based on patient anatomy using a soft material called gel wax. To recreate acoustically realistic tissue properties, two additives to gel wax were considered: paraffin wax to increase acoustic attenuation, and solid glass spheres to increase backscattering. The frequency dependence of ultrasound attenuation was well described with a power law over the measured range of 3-10 MHz. With the addition of paraffin wax in concentrations of 0 to 8 w/w%, attenuation varied from 0.72 to 2.91 dB cm-1 at 3 MHz and from 6.84 to 26.63 dB cm-1 at 10 MHz. With solid glass sphere concentrations in the range of 0.025-0.9 w/w%, acoustic backscattering consistent with a wide range of ultrasonic appearances was achieved. Native gel wax maintained its integrity during compressive deformations up to 60%; its Young's modulus was 17.4 ± 1.4 kPa. The gel wax with additives was shaped by melting and pouring it into 3D printed moulds. Three different phantoms were constructed: a nerve and vessel phantom for peripheral nerve blocks, a heart atrium phantom, and a placental phantom for minimally-invasive fetal interventions. In the first, nerves and vessels were represented as hyperechoic and hypoechoic tubular structures, respectively, in a homogeneous background. The second phantom comprised atria derived from an MRI scan of a patient with an intervening septum and adjoining vena cavae. The third comprised the chorionic surface of a placenta with superficial fetal vessels derived from an image of a post-partum human placenta. Gel wax is a material with widely tuneable ultrasound properties and mechanical characteristics that are well suited for creating patient-specific ultrasound phantoms in several clinical disciplines.

In Vitro Hydrodynamic Assessment of a New Transcatheter Heart Valve Concept (the TRISKELE).

This study presents the in vitro hydrodynamic assessment of the TRISKELE, a new system suitable for transcatheter aortic valve implantation (TAVI), aiming to mitigate the procedural challenges experienced with current technologies. The TRISKELE valve comprises three polymeric leaflet and an adaptive sealing cuff, supported by a novel fully retrievable self-expanding nitinol wire frame. Valve prototypes were manufactured in three sizes of 23, 26, and 29 mm by automated dip-coating of a biostable polymer, and tested in a hydrodynamic bench setup in mock aortic roots of 21, 23, 25, and 27 mm annulus, and compared to two reference valves suitable for equivalent implantation ranges: Edwards SAPIEN XT and Medtronic CoreValve. The TRISKELE valves demonstrated a global hydrodynamic performance comparable or superior to the controls with significant reduction in paravalvular leakage. The TRISKELE valve exhibits enhanced anchoring and improved sealing. The valve is currently under preclinical investigation.

A new transcatheter heart valve concept (the TRISKELE): feasibility in an acute preclinical model.

AIMS: The aim of this study was to introduce and demonstrate the feasibility in an acute preclinical model of a new transcatheter heart valve concept with a self-expanding wire frame, polymeric leaflets and a sealing component. METHODS AND RESULTS: The TRISKELE valve was developed based on a previously validated polymeric leaflet design, an adaptive sealing cuff and a novel nitinol wire frame which reduces stress on the leaflets and radial pressure on the surrounding tissue. A valve prototype of 26 mm nominal diameter was manufactured by automated dip coating of a biostable polymer. The prototype was implanted via brachiocephalic approach in orthotopic position in an acute ovine model through a highly controllable multistage deployment process. The atraumatic retrievability of the valve after full expansion was verified in situ before final release in the optimal position. Observation indicated secure valve anchoring, adequate leaflet motion, and no interference of coronary flow or mitral valve function. CONCLUSIONS: The TRISKELE valve system has the potential to mitigate complications related to imprecise valve positioning, and may offer a safer and more economical TAVI solution to a broad range of patients. The valve is currently under preclinical investigation for its long-term function and durability.

Physical equivalency of wild type and galactose α 1,3 galactose free porcine pericardium; a new source material for bioprosthetic heart valves.

UNLABELLED: Humans make high levels of antibody to carbohydrates with terminal galactose α 1,3 galactose (Gal) modifications. This Gal antigen is widely expressed in other mammals and is present on an array of current animal derived biomedical devices including bioprosthetic heart valves. There is growing interest in using Gal-free animal tissues from Gal knockout pigs (GTKO) as these tissues would not be affected by anti-Gal antibody mediated injury. In this study we compare the composition and biophysical characteristics of glutaraldehyde fixed porcine pericardium from standard and GTKO pigs. We show that with the exception of the Gal antigen which is only present in standard pig tissue both GTKO and standard pig tissue have the same general morphology and collagen content. Moreover uniaxial stress testing and suture retention testing indicate the tissues are equivalent in tensile strength. These studies indicate that genetic disruption of the α-galactosyltransferase (GGTA-1) which blocks synthesis of the Gal antigen has no significant impact on the structural integrity of porcine pericardium and suggest that this tissue could be directly substituted for standard pig pericardium in biomedical devices such as bioprosthetic heart valves. STATEMENT OF SIGNIFICANCE: Surgical heart valve replacement is a proven life saving therapy to treat heart valve dysfunction due to birth defects, infection and the effects of aging. Bioprosthetic heart valves (BHV) made from glutaraldehyde fixed animal tissues are an effective durable therapy in older patients (>60years) but exhibit age-dependent structural valve degeneration (SVD) in younger patients (<60years). SVD is principally caused by BHV calcification. Immune injury contributes to age-dependent SVD through the interaction of galactose α 1,3 galactose (Gal) a dominant xenogeneic antigen present on commercial BHVs and universally abundant human anti-Gal antibody. This study measures the tissue equivalency between standard pig pericardium and Gal-free pericardium from genetically modified pigs as a first step towards making Gal-free BHVs.

Manufacturing and hydrodynamic assessment of a novel aortic valve made of a new nanocomposite polymer.

Synthetic leaflet heart valves have been widely studied as possible alternatives to the current mechanical and bioprosthetic valves. Assessing the in vitro hydrodynamic function of these prostheses is of great importance to predict their hemodynamic behaviour prior to implantation. This study introduces an innovative concept of a low-profile semi-stented surgical aortic valve (SSAV) made of a novel nanocomposite polyurethane with a polycarbonate soft segment (PCU) and polyhedral oligomeric silsesquioxane (POSS) nanoparticles covalently bonded as a pendant cage to the hard segment. The POSS-PCU is already used in surgical implants, including lacrimal duct, bypass graft, and recently, a tracheal replacement. Nine valves of three leaflet thicknesses (100, 150 and 200 µm) and 21 mm internal diameter were prepared using an automated dip-coating procedure, and assessed in vitro for their hydrodynamic performance on a pulse duplicator system. A commercially available porcine bioprosthetic valve (Epic™, St. Jude Medical) of equivalent size was selected as a control model. Compared to the bioprosthetic valve, the SSAVs showed a considerably lower transvalvular pressure drop and larger effective orifice area (EOA). They were also characterised by a lower systolic energy loss, especially at high cardiac outputs. The leaflet thickness was found to significantly affect the hydrodynamics of these valves (P<0.01). The SSAVs with 100 µm leaflets demonstrated improved flow characteristics compared to the bioprosthetic valve. The enhanced hydrodynamic function of the SSAV suggests that the proposed design together with the advanced POSS-PCU material can represent a significant step towards the introduction of polyurethane valves into the clinical application.