bluuebunny/crossref_metadata_2025_split huggingface dataset:
Subcutaneous Testing of E-spun PCL Patches Suitable for in Situ Heart Valve Tissue Engineering
Authors: Talacua Hanna , Vink Aryan , Cox Martijn A.J. , Simonet Marc , Baaijens Frank P.T. , van Herwerden Lex A. , Kluin Jolanda 
Published: July 2012

AbstractIn situ tissue engineered heart valves yields a new generation of cardiovascular substitutes. The body is used as a bioreactor where it relies on the natural regenerative potential of the body. The shift from the classical way of tissue engineering to in-situ tissue engineering emphasizes the role of the scaffold. The scaffold should be able to capture and preserve cells for tissue formation and it has to maintain valve functionality while tissue is developing. The use of synthetic biomaterials is very attractive. Poly(ε-caprolactone) (PCL) is an important polymer due to its mechanical properties and miscibility with a large range of other polymers. Electrospinning attracted great interest as a production method of biomaterials for in situ tissue engineering. The electrospinning process of PCL offers a nice technique for thin fiber formation to eventually create three dimensional scaffolds with the characteristic three layers, typical for heart valves. The fibers produced with electrospinning provide a similar physical structure as the extra cellular matrix. The space between fibers needs to be large enough for cells to adhere and migrate into the scaffold. Sufficient cellular in growth is needed for tissue formation. In this study cellular in growth was measured in subcutaneously implanted electrospun PCL patches. Furthermore tissue formation and degradation of the polymer is investigated. Thirty healthy male F344 Rats are used. The implants with surrounding tissue were explanted after 2, 5, 10, 21 or 84 days and embedded in paraffin. The explanted tissues were examined using immunohistochemistry (HE,MPO, ED1, α-SMA, and PSR). The electrospun fibers had a diameter of 10 μm. SEM pictures showed controlled void spaces. In the electrospun PCL samples we found a very high cellular infiltration rate after 84 days, mean of 396.819 cells per high power field. First infiltration of mainly neutrophil granulocytes was seen, followed by macrophages. Cells infiltrate throughout the whole sample. After 84 days fibroblast were seen, which were able to produce collagen. Furthermore after 84 days, macrophage giant cells and neo-vessel formation was observed. PCL was degraded in the cytoplasm of the macrophages. Electrospun PCL scaffolds with a fiber diameter of 10 μm, are suitable for cellular in growth and tissue formation. This research forms part of the Project P1.01 iValve of the research program of the BioMedical Materials institute, co-funded by the Dutch Ministry of Economic Affairs, Agriculture and Innovation. The financial contribution of the Nederlandse Hartstichting is gratefully acknowledged.
Mechanical Properties of Bioprostheses Leaflets Compared to Human Aortic Valve
Authors: Kalejs Martins , Stradins Peteris , Lacis Lacis , Ozolanta Iveta , Kasyanov Vladimir 
Published: July 2012

AbstractHeart valve bioprostheses suffer from gradual tissue deterioration, which has a causal link with valve tissue mechanical properties. Limited data on mechanical properties of commercially available bioprostheses comparing them to native human aortic valves (AV) is available. Our objective was to determine the mechanical properties of several contemporary bioprostheses and compare them with native human and porcine aortic valves. Leaflets from 5 unchanged human AV, collected from cadaveric hearts and 5 porcine AV, and from 3 of each kind of bioprostheses - Medtronic Hancock II, Sorin Soprano and Medtronic Freestyle were analysed using uniaxial tensile tests in radial and circumferential directions. Data are presented as means ± standard deviation. In both tested directions there's a shift to the stress axis of stress-strain curve for HancockII prostheses and even more for Soprano prostheses when compared to native human valves. In circumferential direction modulus of elasticity (E) of native human AV is 15.34±3.84MPa, porcine AV - 9.7±1.3MPa, Freestyle - 9.0±3.0MPa, HancockII - 22.5±2.2MPa and Soprano - 29.5±6.0MPa. In radial direction E of native human AV is 1.98±0.15MPa, porcine AV - 1.0±0.2MPa, Freestyle - 0.8±0.3MPa, HancockII - 2.5±0.2MPa and Soprano - 15.8±5.4MPa.
Xeno-aortic bioprostheses have a non-linear and anisotropic response to stress in uniaxial tensile tests similar to native AV leaflets. HancockII has gained mechanical strength but lost tissue elasticity compared to native valve tissue. Leaflets of Soprano prostheses are even more rigid and lack pronounced material anisotropy. These differences in mechanical properties may accelerate deterioration of bioprostheses, causing altered stress distribution within valve leaflets. These data provide important information about what mechanical properties future valve substitutes should conform to.
Polymer Nanofiber Materials Matching the Mechanic Properties of Native Aortic Valve
Authors: Kalejs Martins , Stradins Peteris , Priedite Viktorija , Lacis Romans , Ozolanta Iveta , Kasyanov Vladimir 
Published: July 2012

AbstractPorous electrospun nanofiber materials are very promising as matrices for heart valve tissue engineering. Not only biocompatibility is important for this material but also the mechanical features – it has to be strong enough to withhold the pressure after implantation as well as deformable enough for better distribution of shear stress along its surface. Deformability is also crucial for stimulation of fibre production by fibroblasts on these matrices. Altogether 8 differing density variants of electrospun nanofiber materials from gelatine, polyurethane (PUR), polylactic acid (PLA) and polycaprolactone (PCL) were analysed using uniaxial tensile tests. Data were compared to mechanical properties of porcine aortic valve (AV) leaflets in radial and circumferential directions. Data are presented as means ± standard deviation. In circumferential direction modulus of elasticity (E) of native porcine AV is 9.7±1.3MPa and - 1.0±0.2MPa in radial. Ultimate stress and strain is 44.8±5.9% and 2.3±0.6 MPa in circumferential and 95.6±31.4% and 0.5±0.2MPa in radial direction for native leaflets. Closest of the materials to match the mechanical properties of porcine AV in circumferential direction was PUR with density 6.2 g/sqm showing E of 3.9±0.5 MPa, ultimate stress and strain - 5.3±1.68MPa and 141.8±43.9MPa respectively. Closest to match radial direction was gelatine with density 5.7 g/sqm showing E of 0.64±0.14 MPa, ultimate stress and strain - 0.38±0.05MPa and 82.53±10.20MPa respectively.
Native AV leaflets have a non-linear and anisotropic response to stress in uniaxial tensile tests. Hence to model as precisely as possible their mechanical properties we suggest to use a combined material made in a sandwich fashion with layers of gelatine on the outside and PUR in the middle with their fibbers predominantly orientated in perpendicular directions. The other tested materials PLA and PCL either lacked strength to mimic leaflets in circumferential direction or deformability required for the radial direction.
Prevalence of Severe Aortic Stenosis With Low Flow in Patients With and Without Left Ventricular Dysfunction. A Consecutive Echocardiographic Population
Authors: Mauro Toniolo , Andrea Rossi , Mariantonietta Cicoira , Corinna Bergamini , Corrado Vassanelli 
Published: July 2012

AbstractPatients with aortic stenosis (AS) may have a severely reduced aortic valve area (AVA) and a paradoxically low mean gradient (MG). Although this condition is well known in presence of left ventricular (LV) dysfunction, it has recently been observed that it can be associated with normal ejection fraction (EF) as well. Since the prevalence of this condition with respect to LV function is not well defined, we aimed to evaluate the distribution of patients with severely reduced valve area and low MG in a group of patients regardless of EF over a set period of time. We retrospectively identified consecutive patients with severe AS (defined as aortic valve area <0.6 cmq/mq) from our echo data-base. Low MG was defined as < 40 mmHg. Left ventricular systolic dysfunction was considered as EF< 50%. 167 patients with AVA <0.6 cmq/mq formed the study population. 94 (56%) patients were characterized by high MG and 73 (44%) by low MG. Among patients with low MG, 38 were characterized by normal EF and 35 by reduced EF. Differences among groups are shown in Table 1. In this echocardiographic series of consecutive patients, the prevalence of low MG despite severely reduced AVA was high. The distribution of low MG was similar in presence of reduced and preserved EF (21 and 23% of the overall population respectively).
Proteoglycan-Rich Leaflet Thickening in Diet-Induced Early Aortic Valve Disease
Authors: Blaser Mark C. , Sider Krista L. , Kwong Andrea V. , Zhou Yu-Qing , Henkelman R. Mark , Simmons Craig A. 
Published: July 2012

AbstractAortic valve disease (AVD) is a cell-mediated pathology without effective pharmacotherapy, and the early pathogenesis of AVD is drastically understudied. Examining the early stages of disease development may identify biomarkers and novel treatment strategies for use before an untreatable burden of calcification develops. We investigated the effects of a mildly atherogenic diet on early AVD development in mice, and the involvement of proteoglycans (PG) and Sox9 in this process. Male wild-type (WT) C57Bl/6J mice were fed a control diet or BioServ F3282, a high-fat, high-carbohydrate diet (HF/HC) with 58.7% kcal from fat (cholesterol <0.1% w/w) for four months (n = 4-6 per group). Aortic valve function was examined by high-resolution ultrasound biomicroscopy (UBM). Longitudinal aortic valve sections from formalin-fixed and paraffin-embedded hearts were stained by Movat’s pentachrome (morphological changes and ECM composition) and Osteosense 680 (calcification), then immunostained for α-smooth muscle actin (αSMA) and Sox9. Mice on the HF/HC diet for four months became significantly obese (46.7 ± 4.7 vs. 32.3 ± 0.8 g, p < 0.05) but did not develop cardiac hypertrophy. They developed mild but statistically significant hypercholesterolemia (4.7 ± 1.0 vs. 3.1 ± 0.4 mmol/L total cholesterol, p < 0.05). UBM revealed moderate decreases in aortic valve opening area along with increased left ventricular ejection time in HF/HC mice, while no mice exhibited aortic regurgitation. Significant valve thickening was found in the distal third of HF/HC leaflets (84.4 ± 10.4 vs. 37.3 ± 2.7 µm, p < 0.05), while proximal and medial regions were unaltered. Valve thickening was primarily due to PG deposition in HF/HC leaflets (11435 ± 7681 vs. 5448 ± 2948 µm2, p < 0.01), not an increase in collagen content (1729 ± 815 vs. 1771 ± 663 µm2, p = 0.87). Sox9 expression was increased in thickened HF/HC leaflets, and was most highly expressed in PG-rich lesions. HF/HC leaflets did not stain positive for αSMA, implying that changes in leaflet structure and function were not the result of actively synthetic myofibroblasts. Osteosense 680 staining was negative, denoting that the HF/HC diet did not induce leaflet microcalcification.These studies reveal that a high-fat Western diet lacking cholesterol can induce changes in the ECM and functional properties of a WT mouse aortic valve. This early AVD is characterized by thickened leaflets with lesions that are PG-rich and have increased Sox9 expression. Although AVD is considered to be driven by pathological myofibroblast differentiation leading to fibrosis and calcification, the early changes observed herein occur independently of myofibroblast activation, and provide new insight into the initiation and pathogenesis of AVD.
Angiotensin Receptor Blocker Has no Effect on Atherosclerotic Factors in AVS
Authors: Armstrong Zachary B. , Boughner Derek R. , Carruthers Colin P. , Drangova Maria , Rogers Kem A. 
Published: July 2012

AbstractAortic valve sclerosis (AVS) is a chronic progressive disease affecting 25% of the population over the age of 65. Despite this high prevalence, there are currently no preventative therapies which inhibit the progression of AVS. This study sought to determine the effects of an angiotensin II type 1 receptor blocker (ARB), alone or in combination with a statin, on AVS.
Male New Zealand White rabbits were fed either regular chow (Control, n=5) or an atherogenic diet for a period of 18 months to induce AVS. Recognizing the clinical reality, therapy was introduced after disease onset. After 12 months, rabbits were block randomly assigned to four groups receiving either no treatment (Cholesterol, n=6), olmesartan medoxomil (Olmesartan, n=7), atorvastatin calcium (Atorvastatin, n=7), or a combination of both drugs (Combination, n=7) for the final 6 months. Magnetic resonance imaging (MRI) was used to monitor disease progress throughout the treatment period. After sacrifice, valve lesions were analyzed using histology and immunohistochemistry. In vivo disease monitoring yielded no discernible treatment effect. While Cholesterol cusps were significantly thicker than Control throughout the treatment period (0.465 ± 0.030 vs 0.388 ± 0.023mm for Cholesterol and Control, respectively, at 18 months), the various treatments had no positive effect on cusp thickness, and were all identical to Cholesterol at 18 months. Aortic valve area provided similar results; while significant disease was established (0.379 ± 0.033 vs 0.623 ± 0.074cm2 for Cholesterol and Control, respectively, at 18 months), there were no significant differences between treatment groups. Histological analysis of Cholesterol, Atorvastatin, Olmesartan, and Combination cusps revealed fibrosal thickening, lipid deposition, macrophage infiltration, and minor calcification. However, morphological analysis did not reveal significant differences in lesion composition among the treatment groups. Treatment efficacy was confirmed by analysis of aortic lesion area which revealed a significant reduction of atherosclerosis in Olmesartan-treated animals. Neither olmesartan medoxomil nor atorvastatin calcium, alone or in combination, provide demonstrable benefit in the treatment of established AVS despite success in the treatment of atherosclerosis.
Quantification of the Role of Glycosaminoglycans on the Tensile and Viscoelastic Properties of the Aortic Valve Leaflet
Authors: Tseng Hubert , Kim Eric J. , Patrick S. Connell , Grande-Allen K. Jane 
Published: July 2012

AbstractThe spongiosa is the middle of three layers of the aortic valve leaflet and contains the proteoglycan (PG) versican and glycosaminoglycan (GAG) hyaluronan (HA) in large quantities. The resulting versican-HA aggregates allow the layer to retain large quantities of water, giving the layer a gel-like consistency. This layer is the least understood layer of the leaflet as it pertains to its role in valvular function and mechanics. However, the GAGs in this layer are perceived to be important to valve function, as the loss of GAGs in bioprosthetic heart valves coincides with its failure. The functions attributed to these GAGs include: (1) lubricating shear between the outer layers during flexure and tension; and (2) filling large volumes, resisting compression, and dampening shock from valve closure. The lack of understanding of GAGs role in valvular function is due to the difficulty in its isolation from the rest of the leaflet. For our study, rather than isolate the spongiosa, we enzymatically digest GAGs from the spongiosa via hyaluronidase. Previous studies have shown that the complete enzymatic removal of GAGs from heart valve leaflets greatly increases flexural rigidity. In this study, we instead varied the amounts of GAGs in the leaflets and investigated the effects on the tensile properties of the native leaflet; native leaflets in tension have a bilinear stress-strain curve, little hysteresis and minimal relaxation. GAGs were depleted using varying concentrations (0, 1, 2, 5 U/mL) and application times (8, 24 h) to yield a gradient of 4 GAG amounts ranging between 20-40 μg GAG/mg dry weight of native tissue, although with little change in water content. GAG depletion from the spongiosa was also confirmed using Alcian blue staining, which also verified no gross changes to the outer layers. Leaflets in these GAG depletion gradient conditions are in the progress of being mechanically tested to elucidate the effects of GAGs on the tensile elastic and viscoelastic properties of the native leaflet. This work is a component of a broader effort to elucidate the important role of GAGs and PGs in the function of native and diseased valves and in the design of improved valve replacements.
Biomechanical Stimuli Effects on Valve Endothelial Cell Anti-thrombotic Mechanisms
Authors: Balaoing Liezl R. , Post Allison D. , Lin Adam Y. , Nolasco Leticia , Turner Nancy A. , Moake Joel L. , Grande-Allen K. Jane 
Published: July 2012

AbstractBecause of the degeneration and thrombosis in artificial heart valve implants, it is important to understand the anti-thrombotic mechanisms of cardiac valve endothelial cells (VECs). These anti-thrombotic mechanisms can be integrated into poly(ethylene glycol) diacrylate (PEGDA) tissue-engineered heart valve (TEHV) design. This work will study the effects of (1) PEGDA hydrogel stiffness and (2) specific extracellular matrix (ECM) adhesive peptides on VEC phenotype and anti-thrombotic mechanisms. PEGDA 10% (w/v) hydrogels of MWs 3.4 and 20kDa were polymerized to apply different substrate rigidities in VEC culture. Thiol-ene reactions were used to covalently bind laminin- and fibronectin- derived peptides to the acrylate groups on PEGDA hydrogel surfaces. Laminin-derived peptide motif RKRLQVQLSIRT (RKR) and fibronectin adhesive peptide RGD were modified to include an additional cysteine at the end of each sequence, introducing a free thiol to undergo the thiol-ene reaction. Thiol-PEG-fluorescein (SH-PEG-FITC) served as a negative adhesive substrate control. Porcine aortic VECs were seeded onto each of the ECM-hydrogel combinations and cultured for 2, 6, and 10 days. Cell phenotype, adhesion, and proliferation were then assessed. Analysis of specific VEC anti-thrombotic protein regulation is in progress. At each time point, samples will be analyzed for maintenance of VEC phenotype and expression of thrombotic (von Willebrand Factor [VWF]) and anti-thrombotic (VWF cleaving enzyme [ADAMTS-13], eNOS, PGI2, tPA) proteins using histochemistry and qRT-PCR. Addition of histamine has been shown to stimulate rapid release of thrombogenic ultra-large VWF (ULVWF) strings by vascular endothelial cells. This method will be used to study VEC ULVWF string production and the associated cleavage activity of ADAMTS-13. Control of the hemostatic process will be quantified via western blot and ELISAs.
The 3.4 and 20kDa MW PEGDA hydrogels had compressive moduli of 131±5 and 7.5±2kPa, respectively. Binding different concentrations of SH-PEG-FITC onto the gel surfaces showed that acrylate saturation was achieved for both MW compositions using ~5mM of peptide solution. After 2 days, the VECs on the stiffer 3.4kDa RKR gels appeared spread and elongated, whereas the 3.4kDa RGD seeded VECs had cobblestone morphology. VEC adhesion on the RKR and RGD 20kDa gels was observed, but with limited spreading. The cultured VECs may prefer the stiffer 3.4kDa gels over the softer 20kDa gels. After 10 days, VECs on the 3.4kDa RGD gels had minimal proliferation, while VECs on RKR grew confluent, were cobblestone shaped, and expressed VWF. Results suggest that both substrate rigidity and adhesive substrate greatly influence VEC survival, and likely affects anti-thrombotic regulation. Future studies include the use of basal lamina components collagen IV and perlecan peptides to evaluate changes in VEC behavior.
Biomechanical Activation of Human Valvular Interstitial Cells from Early Stage of CAVD
Authors: Poggio Paolo , Sainger Rachana , Grau Juan B. , Branchetti Emanuela , Lai Eric , Gorman Robert C. , Gorman III Joseph H. , Bavaria Joseph E. , Sacks Michael S. , Ferrari Giovanni 
Published: July 2012

AbstractAortic Valve (AV) leaflets are exposed to complex biomechanical stimulations, which has been linked to the progression of Calcific Aortic Valve Disease (CAVD) characterized by valvular interstitial cells (VICs) adopting an osteogenic phenotype, remodeling of the ECM and biomineralization. Initial asymptomatic phases of CAVD, named Aortic Valve Sclerosis (AVSc), include thickening of the cusps whereas advanced stages, Aortic Valve Stenosis (AVS), are associated with severe calcification. Prospective clinical studies of CAVD are hampered by the typically slow and variable progression of the disease. In addition, patients who present with AVS are already in the stage of severe calcification where damage to the AV leaflets is too severe to be reversed by drug therapy. Finally, due to its asymptomatic presentation little is known about the AVSc stage. The aim of this study is to develop tissue- and cell-based models to uncover the biomechanical forces leading to activation of VICs in early, asymptomatic AVSc patients. We investigate the impact of biological (BMP4 pathway) and mechanical (tensile stretch) forces leading to VICs osteogenic transdifferentiation, and biomineralization of the fibrosa layer.
Human, surgically resected, AV tissue from AVSc patients were characterized for cellular and extracellular markers and compared to healthy controls and AVS tissues. BMP4 and tensile stretch induce osteogenic marker expression and biomineralization of the fibrosa in non-calcified AVSc-derived tissue. VICs were isolated from patients and induced to transdifferentiate in either 2D cell culture or 3D Tissue Engineering valve model based on decellularized porcine AV scaffold repopulated with human-derived cells. Our results show that the synergistic combination of biological (BMP4) and mechanical (tensile stretch) forces is required to promote SMA, RUNX2, OPN, and ON expression of AVSc-derived cells. These results provide a novel study model of early asymptomatic AVSc that combine biological and mechanical stimulation to induce activation of AVSc-derived VICs and biomineralization of AVSc tissue. This model could be used to unveil the molecular mechanisms lading to VIC activation and to test possible future small drug to control cellular activation and tissue remodeling.