Negative In Vivo Results Despite Promising In Vitro Data With a Coated Compliant Electrospun Polyurethane Vascular Graft.

INTRODUCTION: There is a growing need for small-diameter (<6 mm) off-the-shelf synthetic vascular conduits for different surgical bypass procedures, with actual synthetic conduits showing unacceptable thrombosis rates. The goal of this study was to build vascular grafts with better compliance than standard synthetic conduits and with an inner layer stimulating endothelialization while remaining antithrombogenic. METHODS: Tubular vascular conduits made of a scaffold of polyurethane/polycaprolactone combined with a bioactive coating based on chondroitin sulfate (CS) were created using electrospinning and plasma polymerization. In vitro testing followed by a comparative in vivo trial in a sheep model as bilateral carotid bypasses was performed to assess the conduits' performance compared to the actual standard. RESULTS: In vitro, the novel small-diameter (5 mm) electrospun vascular grafts coated with chondroitin sulfate (CS) showed 10 times more compliance compared to commercial expanded polytetrafluoroethylene (ePTFE) conduits while maintaining adequate suturability, burst pressure profiles, and structural stability over time. The subsequent in vivo trial was terminated after electrospun vascular grafts coated with CS showed to be inferior compared to their expanded polytetrafluoroethylene counterparts. CONCLUSIONS: The inability of the experimental conduits to perform well in vivo despite promising in vitro results may be related to the low porosity of the grafts and the lack of rapid endothelialization despite the presence of the CS coating. Further research is warranted to explore ways to improve electrospun polyurethane/polycaprolactone scaffold in order to make it prone to transmural endothelialization while being resistant to strenuous conditions.

Combining Electrospun Fiber Mats and Bioactive Coatings for Vascular Graft Prostheses.

The patency of small-diameter (<6 mm) synthetic vascular grafts (VGs) is still limited by the absence of a confluent, blood flow-resistant monolayer of endothelial cells (ECs) on the lumen and of vascular smooth muscle cell (VSMC) growth into the media layer. In this research, electrospinning has been combined with bioactive coatings based on chondroitin sulfate (CS) to create scaffolds that possess optimal morphological and bioactive properties for subsequent cell seeding. We fabricated random and aligned electrospun poly(ethylene terephthalate), ePET, mats with small pores (3.2 ± 0.5 or 3.9 ± 0.3 µm) and then investigated the effects of topography and bioactive coatings on EC adhesion, growth, and resistance to shear stress. Bioactive coatings were found to dominate the cell behavior, which enabled creation of a near-confluent EC monolayer that resisted physiological shear-flow conditions. CS is particularly interesting since it prevents platelet adhesion, a key issue to avoid blood clot formation in case of an incomplete EC monolayer or partial cell detachment. Regarding the media layer, circumferentially oriented nanofibers with larger pores (6.3 ± 0.5 µm) allowed growth, survival, and inward penetration of VSMCs, especially when the CS was further coated with tethered, oriented epithelial growth factor (EGF). In summary, the techniques developed here can lead to adequate scaffolds for the luminal and media layers of small-diameter synthetic VGs.

Is there a relationship between cardiorespiratory fitness and executive performance as function of mental workload in young adults?

In the current study, we have decided to investigate the relationship between cardiorespiratory fitness and executive functions in young adults as a function of mental workload. To achieve our objectives, we have solicited 29 young adults (18-25 years; 12 women) who have first realized the Random Number Generation (RNG) task with two levels of complexity. After each level of complexity, the participants were asked to report on their perceived difficulty. Secondly, participants performed the RABIT® test, during which oxygen consumption was measured through the Metamax 3B-R2. The results showed that executive performance and perceived difficulty deteriorate with increasing task complexity. Additionally, oxygen consumption increased significantly to reach a peak during the hardest phase of the RABIT® test. Finally, as in previous studies, we could not observe a correlation between cardiorespiratory fitness and executive functions. Our findings support the lack of a direct relationship between cardiorespiratory fitness and executive functions. Future studies should explore the relationship between the composite measure of executive function, hemodynamic activity, and cardiorespiratory fitness in healthy youth and their peers with cardiovascular disease. This will examine an indirect effect of Cardiorespiratory fitness (CRF) on Executive functions (EFs) through brain activity.

Towards compliant small-diameter vascular grafts: Predictive analytical model and experiments.

The search for novel, more compliant vascular grafts for the replacement of blood vessels is ongoing, and predictive tools are needed to identify the most promising biomaterials. A simple analytical model was designed that enables the calculation of the ratio between the ultimate stress (σult) and the elastic modulus (E). To reach both the compliance of small-diameter coronary arteries (0.0725%/mmHg) and a burst pressure of 2031 mmHg, a material with a minimum σult/E ratio of 1.78 is required. Based on this result and on data from the literature, random electrospun Polyurethane/Polycaprolactone (PU/PCL) tubular scaffolds were fabricated and compared to commercial ePTFE prostheses. PU/PCL grafts showed mechanical properties close to those of native arteries, with a circumferential elastic modulus of 4.8 MPa and a compliance of 0.036%/mmHg at physiological pressure range (80-120 mmHg) for a 145 µm-thick prosthesis. In contrast, commercial expanded polytetrafluoroethylene (ePTFE) grafts presented a high Young's modulus (17.4 MPa) and poor compliance of 0.0034%/mmHg. The electrospun PU/PCL did not however reach the target values as its σult/E ratio was lower than expected, at 1.54, well below the calculated threshold (1.78). The model tended to overestimate both the compliance and burst pressure, with the differences between the analytical and experimental results ranging between 13 and 34%, depending on the pressure range tested. This can be explained by the anisotropy of random electrospun PU/PCL and its slightly non-linear elastic behavior, in contrast to the hypotheses of our model. Impermeability tests showed that the electrospun scaffolds were impermeable to blood for all thicknesses above 50 µm. In conclusion, this analytical model allows to select materials with suitable mechanical properties for the design of small-diameter vascular grafts. The novel electrospun PU/PCL tubular scaffolds showed strongly improved compliance as compared to commercial ePTFE prostheses.

Retention of transforming growth factor beta1 using functionalized dextran-based hydrogels.

Functionalized dextrans (FD) are anionic water-soluble polymers bearing carboxylate, benzylamide and sulfate groups, which exhibit binding capacity to transforming growth factor-beta1 (TGF-beta1). In this paper, we have investigated the ability of dextran-based hydrogels containing FD, to bind and release recombinant human TGF-beta1. Hydrogels were prepared by chemical crosslink native dextran and FD with sodium trimetaphosphate in 1m NaOH at 50 degrees C. A wide range of hydrogels were prepared as particles ranging of 1-1.6mm of diameter and characterized with various amounts of FD and with different crosslinker feeding ratios (CFR). Dried particles were soaked with recombinant human transforming growth factor-beta1 (rhTGF-beta1) to determine their capacity to deliver the growth factor. Results indicated that the in vitro kinetics releases of rhTGF-beta1 were related to FD and CFR. Retention capacity of rhTGF-beta1 increases with an increase of negative charges of the matrices brought by both phosphate linkages and FD as demonstrated by an additional release of growth factor in high ionic strength solution. Highly crosslinked hydrogels that contained the highest amount of FD (18% (w/w)) retained up to 88% of rhTGF-beta1. Bioactivity of released growth factor was confirmed in a cell assay. These functionalized hydrogels may have important uses for the stabilization and the protection of rhTGF-beta1 as entrapment systems and could be applied to other proteins of clinical interest.

Bovine BMP osteoinductive potential enhanced by functionalized dextran-derived hydrogels.

This study evaluated functionalized dextran-derived hydrogels as BMP carriers using both in vitro and in vivo models. In vitro release kinetics indicated that dextran-derived hydrogels could retain rhBMP-2 growth factor in a variable manner depending on their functionalization ratio. The potential of these hydrogels when combined with extracted bovine BMP to enhance the bone formation was evaluated in a rat ectopic model. The largest osteoinduction was found when using hydrogels exhibiting the highest growth factor retention capacity. In addition, some implanted hydrogels demonstrated a capacity to induce an in-vivo calcification certainly related to their chemical composition. These properties make these materials interesting osteoconductive BMP carriers, allowing to decrease the amount of implanted factor required for bone regeneration.

Injectable thermosensitive chitosan hydrogels with controlled gelation kinetics and enhanced mechanical resistance.

The use of injectable hydrogels is presently limited by the difficulty to achieve rapid gelation, high mechanical resistance and excellent cytocompatibility. In our study, high-strength injectable thermosensitive hydrogels of unmodified chitosan (CH) were obtained by combining sodium hydrogen carbonate (SHC) with phosphate buffer (PB) or beta-glycerophosphate (BGP) as gelling agents. A synergic effect led to the acceleration of gelation and a remarkable improvement of the storage modulus (G') of the hydrogels. Furthermore, the new hydrogels exhibited drastically enhanced Young moduli and resistance in compression as compared to conventional hydrogels prepared with BGP, PB or their combination. This was achieved while reducing the total salt concentration in the hydrogels. The gelation was rapid and the hydrogels presented porous structures and physiological pH, and did not show any cytotoxicity to L929 fibroblast cells in vitro. Overall, these new hydrogels provide interesting alternatives for use as blood vessel embolizing agents or as injectable scaffolds for drug delivery and/or cell seeding in tissue engineering strategies.

Electrospun nanofiber scaffolds and plasma polymerization: a promising combination towards complete, stable endothelial lining for vascular grafts.

In the quest to reduce risk of thrombosis in vascular grafts, it is essential to provide a surface with morphological and mechanical properties close to those of the extracellular matrix beneath the luminal endothelium, and to favor the growth of a confluent, stable monolayer of endothelial cells. This is accomplished here by combining electrospun poly(ethylene terephthalate) (PET) mats with an amine-rich thin plasma-polymerized coating, designated "L-PPE:N." Its deposition does not modify the open, highly porous mats and leads only to small changes in mechanical properties. L-PPE:N significantly improves the adhesion and growth of human umbilical vein endothelial cells (HUVEC) and their resistance to flow-induced shear stress. These properties favor the formation of desired confluent HUVEC monolayers on the topmost surface, unlike conventional vascular grafts (ePTFE or woven PET), where cells migrate inside the material. This combination is therefore highly advantageous for the pre-endothelialization of the luminal side of small-diameter vascular prostheses.

Low thrombogenicity coating of nonwoven PET fiber structures for vascular grafts.

Vascular PET grafts (Dacron) have shown good performance in large vessels (≥ 6 mm) applications. To address the urgent unmet need for small-diameter (2-6 mm) vascular grafts, proprietary high-compliance nonwoven PET fiber structures were modified with various PEG concentrations using PVA as a cross-linking agent, to fabricate non-thrombogenic mechanically compliant vascular grafts. The blood compatibility assays measured through platelet adhesion (SEM and mepacrine dye) and platelet activation (morphological changes, P-selectin secretion, and TXB2 production) demonstrate that functionalization using a 10% PEG solution was sufficient to significantly reduce platelet adhesion/activation close to optimal literature-reported levels observed on carbon-coated ePTFE.