Multi-layer approaches to scaffold-based small diameter vessel engineering: A review.

Cardiovascular disease is one of the leading causes of death in the world. A characteristic symptom of cardiovascular disease is occlusion of vessels. Once vascular occlusion occurs there is a critical need to re-establish flow to prevent ischemia in the downstream tissues. In the most advanced cases, flow is re-established by creating a secondary flow path around the blockage, bypass grafting. For large diameter applications, synthetic conduits are successfully implanted, however in small diameter applications re-occlusion occurs and there is a critical need for new vascular grafts. There are many strategies and approaches that are being employed to design an effective and successful vascular graft. However, to date, there are no clinically available small diameter vascular grafts that are consistently successful in vivo long term (>7 years). As an effort to develop a successful graft there are several tissue engineering approaches: cell sheets, synthetic and natural biomaterial platforms, and decellularized extracellular matrices that are being investigated. While each area has its advantages, scaffold-based approaches are among the most widely studied. Scaffold based approaches are extensively studied due to tailorability and the availability of synthetic and natural polymers. Within the area of scaffold-based approaches, biomimicry has become an increasingly studied area, and structural biomimicry is one of the many approaches. The focus of this review paper is to analyze scaffold-based approaches. Particularly the advantages and disadvantages of using multi-layer scaffold-based approaches to engineer conduits for small diameter applications.

Periadventitial atRA citrate-based polyester membranes reduce neointimal hyperplasia and restenosis after carotid injury in rats.

Oral all-trans retinoic acid (atRA) has been shown to reduce the formation of neointimal hyperplasia; however, the dose required was 30 times the chemotherapeutic dose, which already has reported side effects. As neointimal formation is a localized process, new approaches to localized delivery are required. This study assessed whether atRA within a citrate-based polyester, poly(1,8 octanediolcitrate) (POC), perivascular membrane would prevent neointimal hyperplasia following arterial injury. atRA-POC membranes were prepared and characterized for atRA release via high-performance liquid chromatography with mass spectrometry detection. Rat adventitial fibroblasts (AF) and vascular smooth muscle cells (VSMC) were exposed to various concentrations of atRA; proliferation, apoptosis, and necrosis were assessed in vitro. The rat carotid artery balloon injury model was used to evaluate the impact of the atRA-POC membranes on neointimal formation, cell proliferation, apoptosis, macrophage infiltration, and vascular cell adhesion molecule 1 (VCAM-1) expression in vivo. atRA-POC membranes released 12 µg of atRA over 2 wk, with 92% of the release occurring in the first week. At 24 h, atRA (200 µmol/l) inhibited [(3)H]-thymidine incorporation into AF and VSMC by 78% and 72%, respectively (*P = 0.001), with negligible apoptosis or necrosis. Histomorphometry analysis showed that atRA-POC membranes inhibited neointimal formation after balloon injury, with a 56%, 57%, and 50% decrease in the intimal area, intima-to-media area ratio, and percent stenosis, respectively (P = 0.001). atRA-POC membranes had no appreciable effect on apoptosis or proliferation at 2 wk. Regarding biocompatibility, we found a 76% decrease in macrophage infiltration in the intima layer (P < 0.003) in animals treated with atRA-POC membranes, with a coinciding 53% reduction in VCAM-1 staining (P < 0.001). In conclusion, perivascular delivery of atRA inhibited neointimal formation and restenosis. These data suggest that atRA-POC membranes may be suitable as localized therapy to inhibit neointimal hyperplasia following open cardiovascular procedures.

Effect of bilateral oophorectomy on wound healing of the rabbit vagina.

We aimed to assess the effect of bilateral oophorectomy on vaginal wound healing in three groups of New Zealand White rabbits (24 rabbits each). Group 1 underwent bilateral oophorectomy, group 2 underwent a sham surgery, and group 3 served as control. Standardized vaginal tissue specimens were harvested and assessed for wound and scar surface area and tensiometric analysis before wounding, and sequentially thereafter, showing that vaginal wound closure, scar contraction, and recovery of biomechanical properties are significantly slower in oophorectomized rabbits.

Citric acid-based elastomers provide a biocompatible interface for vascular grafts.

Prosthetic vascular bypass grafting is associated with poor long-term patency rates. Herein, we report on the mid-term performance of expanded polytetrafluoroethylene (ePTFE) vascular grafts modified with a citric acid-based biodegradable elastomer. Through a spin-shearing method, ePTFE grafts were modified by mechanically coating a layer of poly(1,8 octanediol citrate) (POC) onto the luminal nodes and fibrils of the ePTFE. Control and POC-ePTFE grafts were implanted into the porcine carotid artery circulation as end-to-side bypass grafts. Grafts were assessed by duplex ultrasonography, magnetic resonance angiography, and digital subtraction contrast angiography and were all found to be patent with no hemodynamically significant stenoses. At 4 weeks, POC-ePTFE grafts were found to be biocompatible and resulted in a similar extent of neointimal hyperplasia as well as leukocyte and monocyte/macrophage infiltration as control ePTFE grafts. Furthermore, POC supported endothelial cell growth. Lastly, scanning electron microscopy confirmed the presence of POC on the ePTFE grafts at 4 weeks. Thus, these data reveal that surface modification of blood-contacting surfaces with POC results in a biocompatible surface that does not induce any untoward effects or inflammation in the vasculature. These findings are important as they will serve as the foundation for the development of a drug-eluting vascular graft.

In vitro characterization of a compliant biodegradable scaffold with a novel bioreactor system.

The influence of scaffold compliance on blood vessel tissue engineering remains unclear and compliance mismatch issues are important to an in vivo tissue-engineering approach. We have designed and constructed a modular bioreactor system that is capable of imparting pulsatile fluid flow while simultaneously measuring vessel distension with fluid pressure changes in real time. The setup uses a pneumatic PID control system to generate variable fluid pressure profiles via LabVIEW and an LED micrometer to monitor vessel distension to an accuracy of +/-2 microm. The bioreactor was used to measure the compliance of elastomeric poly(1,8-octanediol citrate) (POC) scaffolds over physiologically relevant pressure ranges. The compliance of POC scaffolds could be adjusted by changing polymerization conditions resulting in scaffolds with compliance values that ranged from 3.8 +/- 0.2 to 15.6 +/- 4.6%/mmHg x 10(-2), depending on the distension pressures applied. Furthermore, scaffolds that were incubated in phosphate-buffered saline for 4 weeks exhibited a linear increase in compliance (2.6 +/- 0.9 to 7.7 +/- 1.2%/mmHg x 10(-2)) and were able to withstand normal physiological blood pressure without bursting. The ability to tailor scaffold compliance and easily measure vessel compliance in real time in vitro will improve our understanding of the role of scaffold compliance on vascular cell processes.

Hemocompatibility evaluation of poly(glycerol-sebacate) in vitro for vascular tissue engineering.

Poly(glycerol-sebacate) (PGS) is an elastomeric biodegradable polyester that could potentially be used to engineer blood vessels in vivo. However, its blood-material interactions are unknown. The objectives of this study were to: (a) fabricate PGS-based biphasic tubular scaffolds and (b) assess the blood compatibility of PGS in vitro in order to get some insight into its potential use in vivo. PGS was incorporated into biphasic scaffolds by dip-coating glass rods with PGS pre-polymer. The thrombogenicity (platelet adhesion and aggregation) and inflammatory potential (IL-1beta and TNFalpha expression) of PGS were evaluated using fresh human blood and a human monocyte cell line (THP-1). The activation of the clotting system was assessed via measurement of tissue factor expression on THP-1 cells, plasma recalcification times, and whole blood clotting times. Glass, tissue culture plastic (TCP), poly(l-lactide-co-glycolide) (PLGA), and expanded polytetrafluorethylene (ePTFE) were used as reference materials. Biphasic scaffolds with PGS as the blood-contacting surface were successfully fabricated. Relative to glass (100%), platelet attachment on ePTFE, PLGA and PGS was 61%, 100%, and 28%, respectively. PGS elicited a significantly lower release of IL-1beta and TNFalpha from THP-1 cells than ePTFE and PLGA. Similarly, relative to all reference materials, tissue factor expression by THP-1 cells was decreased when exposed to PGS. Plasma recalcification and whole blood clotting profiles of PGS were comparable to or better than those of the reference polymers tested.

Biomechanical characterization of vaginal versus abdominal surgical wound healing in the rabbit.

OBJECTIVE: The objective of the study was to compare biomechanical properties of vaginal versus abdominal surgical wound healing in the rabbit. STUDY DESIGN: Bilateral 6-mm full-thickness circular segments were excised from the vagina and abdominal skin in 38 New Zealand White female rabbits. Animals were killed 0, 4, 7, 10, 14, 21, 28, and 35 days after wounding, and the wounds were assessed for surface area and tensile properties. RESULTS: Wound closure and scar contraction were significantly faster in the vagina than the abdomen (P = .001). Baseline tensile strength (P = .05), modulus (P = .001), and tensile energy to break (P = .18) were higher in the abdomen, whereas maximal tissue elongation was higher in the vagina (P = .13). After wounding, a drop in tensile strength, modulus, and tensile energy to break was observed in both tissues through postwounding day 4, followed by a progressive recovery of these properties. A progressive loss of elasticity was noted in both tissues after wounding. CONCLUSION: The surgical wound closes and contracts faster in the rabbit vagina than in its abdomen. Although these tissues have different biomechanical properties at baseline, they regenerate their tensile strength and lose their elasticity at a similar rate.

Synthesis and evaluation of poly(diol citrate) biodegradable elastomers.

Herein, we report the synthesis and evaluation of a novel family of biodegradable and elastomeric polyesters, poly(diol citrates). Poly(diol citrates) were synthesized by reacting citric acid with various diols to form a covalent cross-linked network via a polycondensation reaction without using exogenous catalysts. The tensile strength of poly(diol citrates) were as high as 11.15+/-2.62 MPa and Young's modulus ranged from 1.60+/-0.05 to 13.98+/-3.05 MPa under the synthesis conditions that were investigated. Elongation was as high as 502+/-16%. No permanent deformation was found during mechanical tests. The equilibrium water-in-air contact angles of measured poly(diol citrates) films ranged from 15 degrees to 53 degrees . The mechanical properties, degradation and surface characteristics of poly(diol citrates) could be controlled by choosing different diols as well as by controlling the cross-link density of the polyester network. Various types of poly(diol citrate) scaffolds were fabricated to demonstrate their processing potential. These scaffolds were soft and could recover from deformation. In vitro and in vivo evaluation using cell culture and subcutaneous implantation, respectively, confirmed cell and tissue compatibility. The introduction of poly(diol citrates) will expand the repertoire of currently available biodegradable polymeric elastomers and should help meet the requirements of tissue engineering applications.

Novel biphasic elastomeric scaffold for small-diameter blood vessel tissue engineering.

Compliance mismatch, thrombosis, and long culture times in vitro remain important challenges to the clinical implementation of a tissue-engineered small-diameter blood vessel (SDBV). To address these issues, we are developing an implantable elastomeric and biodegradable biphasic tubular scaffold. The scaffold design uses connected nonporous and porous phases as a basis to mimic, respectively, the intimal and medial layers of a blood vessel. Biphasic scaffolds were fabricated from poly(diol citrate), a novel class of biodegradable polyester elastomer. Scaffolds were characterized for tensile and compressive properties, burst pressure, compliance, foreign body reaction (via subcutaneous implantation in rats), and cell distribution and differentiation (via histology, scanning electron microscopy, and immunohistochemistry). Tensile tests, burst pressure, and compliance measurements confirm that the incorporation of a nonporous phase to create a "skin" connected to the porous phase of a scaffold can provide bulk mechanical properties that are similar to those of a native vessel. Compression tests confirm that the scaffolds are soft and recover from deformation. Subcutaneously implanted poly(diol citrate) porous scaffolds produce a thin fibrous capsule and allow for tissue ingrowth. In vitro culture of tubular biphasic scaffolds seeded with human aortic smooth muscle cells (HASMCs) and endothelial cells (HAECs) demonstrates the ability of this design to support cell compartmentalization, coculture, and cell differentiation. The newly formed HAEC monolayer stained positive for von Willebrand factor whereas collagen- and calponin-positive HASMCs were present in the porous phase.

Biodegradable polyester elastomers in tissue engineering.

Tissue engineering often makes use of biodegradable scaffolds to guide and promote controlled cellular growth and differentiation in order to generate new tissue. There has been significant research regarding the effects of scaffold surface chemistry and degradation rate on tissue formation and the importance of these parameters is widely recognised. Nevertheless, studies describing the role of mechanical stimuli during tissue development and function suggest that the mechanical properties of the scaffold will also be important. In particular, scaffold mechanics should be taken into account if mechanical stimulation, such as cyclic strain, will be incorporated into strategies to grow improved tissues or the target tissue to be replaced has elastomeric properties. Biodegradable polyesters, such as polyglycolide, polylactide and poly(lactide-co-glycolide), although commonly used in tissue engineering, undergo plastic deformation and failure when exposed to long-term cyclic strain, limiting their use in engineering elastomeric tissues. This review will cover the latest advances in the development of biodegradable polyester elastomers for use as scaffolds to engineer tissues, such as heart valves and blood vessels.