Jeremy Goldman

Zinc Exhibits Ideal Physiological Corrosion Behavior for Bioabsorbable Stents

Zinc is proposed as an exciting new biomaterial for use in bioabsorbable cardiac stents. Not only is zinc a physiologically relevant metal with behavior that promotes healthy vessels, but it combines the best behaviors of both current bioabsorbable stent materials: iron and magnesium. Shown here is a composite image of zinc degradation in a murine (rat) artery.

Cooperative and redundant roles of VEGFR-2 and VEGFR-3 signaling in adult lymphangiogenesis

Activation of vascular endothelial growth factor (VEGF) receptor‐3 (VEGFR‐3) by VEGF‐C initiates lymphangiogenesis by promoting lymphatic proliferation and migration. However, it is unclear whether VEGFR‐3 signaling is required beyond these initial stages, namely during the organization of new lymphatic endothelial cells (LECs) into functional capillaries. Furthermore, the role of VEGFR‐2, which is also expressed on LECs and binds VEGF‐C, is unclear. We addressed these questions by selectively neutralizing VEGFR‐3 and/or VEGFR‐2 for various time periods in an adult model of lymphangiogen‐esis in regenerating skin. While blocking either VEGFR‐2 or VEGFR‐3 with specific antagonist mAbs (DC101 and mF4–31C1, respectively) prior to lymphatic migration prevented lymphangiogenesis, blocking VEGFR‐3 subsequent to migration did not affect organization into functional capillaries, and VEGFR‐2 blocking had only a small hindrance on organization. These findings were confirmed in vitro using human LECs and anti‐human antagonist mAbs (IMC‐1121a and hF4–3C5): both VEGFR‐2 and ‐3 signaling were required for migration and proliferation, but tubulogenesis in 3D cultures was unaffected by VEGFR‐3 blocking and partially hindered by VEGFR‐2 blocking. Furthermore, both in vitro and in vivo, while VEGFR‐3 blocking had no effect on LEC organization, coneutralization of VEGFR‐2, and VEGFR‐3 completely prevented lymphatic organization. Our findings demonstrate that cooperative signaling of VEGFR‐2 and ‐3 is necessary for lymphatic migration and proliferation, but VEGFR‐3 is redundant with VEGFR‐2 for LEC organization into functional capillaries.—Goldman, J., Rutkowski, J. M., Shields, J. D., Pasquier, M. C., Cui, Y., Schmökel, H. G., Willey, S., Hicklin, D. J., Pytowski, B., Swartz, M. A. Cooperative and redundant roles of VEGFR‐2 and VEGFR‐3 signaling in adult lymphangiogenesis. FASEB J. 21, 1003–1012 (2007)

Overexpression of VEGF-C Causes Transient Lymphatic Hyperplasia but Not Increased Lymphangiogenesis in Regenerating Skin

Vascular endothelial growth factor (VEGF)-C is necessary for lymphangiogenesis and holds potential for lymphangiogenic therapy in diseases lacking adequate lymphatic drainage. However, the ability of VEGF-C to enhance sustainable, functional lymphatic growth in adult tissues remains unclear. To address this, we evaluated VEGF-C overexpression in adult lymphangiogenesis in regenerating skin. We used a model of mouse tail skin regeneration incorporating a suspension of either VEGF-C overexpressing tumor cells, which provide a continuous supplement of excess VEGF-C to the natural regenerating environment for more than 25 days, or otherwise identical control-transfected tumor cells. We found that excess VEGF-C did not enhance the rate of lymphatic endothelial cell (LEC) migration, the density of lymphatic vessels, or the rate of functionality - even though lymphatic hyperplasia was present early on. Furthermore, the hyperplasia disappeared when VEGF-C levels diminished, which occurred after 25 days, rendering the lymphatics indistinguishable from those in control groups. In vitro, we showed that whereas cell-derived VEGF-C could induce chemoattraction of LECs across a membrane (which involves amoeboid-like transmigration), it did not increase LEC chemoinvasion within a 3-dimensional fibrin matrix (which requires proteolytic migration). These results suggest that whereas excess VEGF-C may enhance early LEC proliferation and cause lymphatic vessel hyperplasia, it does not augment the physiological rate of migration or functionality, and by itself cannot sustain any lasting effects on lymphatic size, density, or organization in regenerating adult skin.

A simplified in vivo approach for evaluating the bioabsorbable behavior of candidate stent materials

Metal stents are commonly used to revascularize occluded arteries. A bioabsorbable metal stent that harmlessly erodes away over time may minimize the normal chronic risks associated with permanent implants. However, there is no simple, low-cost method of introducing candidate materials into the arterial environment. Here, we developed a novel experimental model where a biomaterial wire is implanted into a rat artery lumen (simulating bioabsorbable stent blood contact) or artery wall (simulating bioabsorbable stent matrix contact). We use this model to clarify the corrosion mechanism of iron (≥99.5 wt %), which is a candidate bioabsorbable stent material due to its biocompatibility and mechanical strength. We found that iron wire encapsulation within the arterial wall extracellular matrix resulted in substantial biocorrosion by 22 days, with a voluminous corrosion product retained within the vessel wall at 9 months. In contrast, the blood-contacting luminal implant experienced minimal biocorrosion at 9 months. The importance of arterial blood versus arterial wall contact for regulating biocorrosion was confirmed with magnesium wires. We found that magnesium was highly corroded when placed in the arterial wall but was not corroded when exposed to blood in the arterial lumen for 3 weeks. The results demonstrate the capability of the vascular implantation model to conduct rapid in vivo assessments of vascular biomaterial corrosion behavior and to predict long-term biocorrosion behavior from material analyses. The results also highlight the critical role of the arterial environment (blood vs. matrix contact) in directing the corrosion behavior of biodegradable metals.

Metallic zinc exhibits optimal biocompatibility for bioabsorbable endovascular stents

Although corrosion resistant bare metal stents are considered generally effective, their permanent presence in a diseased artery is an increasingly recognized limitation due to the potential for long-term complications. We previously reported that metallic zinc exhibited an ideal biocorrosion rate within murine aortas, thus raising the possibility of zinc as a candidate base material for endovascular stenting applications. This study was undertaken to further assess the arterial biocompatibility of metallic zinc. Metallic zinc wires were punctured and advanced into the rat abdominal aorta lumen for up to 6.5months. This study demonstrated that metallic zinc did not provoke responses that often contribute to restenosis. Low cell densities and neointimal tissue thickness, along with tissue regeneration within the corroding implant, point to optimal biocompatibility of corroding zinc. Furthermore, the lack of progression in neointimal tissue thickness over 6.5months or the presence of smooth muscle cells near the zinc implant suggest that the products of zinc corrosion may suppress the activities of inflammatory and smooth muscle cells.

Role of blood shear stress in the regulation of vascular smooth muscle cell migration

Smooth muscle cell (SMC) migration from the media to the intima of blood vessels contributes to neointimal formation and atherogenesis. Here, the authors demonstrate how blood shear stress regulates vascular SMC migration in the encapsulating tissue of a micro-cylinder implanted in the center of the rat vena cava with the micro-cylinder perpendicular to blood flow, in this model, the micro-cylinder was exposed to a laminar flow with a known shear stress field in the leading region and a vortex flow in the trailing region. After surgery, the micro-cylinder was encapsulated by a thrombus-like tissue within one day, followed by SMC migration from the vena cava to the encapsulating tissue from day 3 to 20. SMC migration was time-dependent with a peak migration speed at day 5. At each given time (excluding day 1), blood shear stress exerts an inhibitory effect on SMC migration with significantly suppressed SMC migration in the laminar flow region than in the stagnation, separation, and vortex flow regions. SMCs were relatively parallel to the shear stress direction in high shear stress regions, whereas perpendicular to the shear stress direction in low shear stress regions. These results suggest that blood shear stress plays a role in regulating SMC migration and orientation in this model.

Magnesium in the murine artery: Probing the products of corrosion

Many publications are available on the physiological and pseudophysiological corrosion of magnesium and its alloys for bioabsorbable implant application, yet few focus on the characterization of explanted materials. In this work, commercially pure magnesium wires were corroded in the arteries of rats for up to 1 month, removed, and both bulk and surface products characterized. Surface characterization using infrared spectroscopy revealed a duplex structure comprising heavily magnesium-substituted hydroxyapatite that later transformed into an A-type (carbonate-substituted) hydroxyapatite. To explain this transformation, an ion-exchange mechanism is suggested. Elemental mapping of the bulk products of biocorrosion revealed the elemental distribution of Ca, P, Mg and O in the outer and Mg, O and P in the inner layers. Carbon was not observed in any significant quantity from the inner corrosion layer, suggesting that carbonates are not a prevalent product of corrosion. Backscatter electron imaging of cross-sections showed that thinning or absence of the hydroxyapatite in the later stages of degradation is related to local thickening of the inner corrosion layer. Based on these experimental observations, mechanisms describing corrosion in the quasi-steady state and during terminal breakdown of the magnesium specimens are proposed.

Recent Advances in Biodegradable Metals for Medical Sutures: A Critical Review

Sutures that biodegrade and dissolve over a period of several weeks are in great demand to stitch wounds and surgical incisions. These new materials are receiving increased acceptance across surgical procedures whenever permanent sutures and long-term care are not needed. Unfortunately, both inflammatory responses and adverse local tissue reactions in the close-to-stitching environment are often reported for biodegradable polymeric sutures currently used by the medical community. While bioabsorbable metals are predominantly investigated and tested for vascular stent or osteosynthesis applications, they also appear to possess adequate bio-compatibility, mechanical properties, and corrosion stability to replace biodegradable polymeric sutures. In this Review, biodegradable alloys made of iron, magnesium, and zinc are critically evaluated as potential materials for the manufacturing of soft and hard tissue sutures. In the case of soft tissue closing and stitching, these metals have to compete against currently available degradable polymers. In the case of hard tissue closing and stitching, biodegradable sternal wires could replace the permanent sutures made of stainless steel or titanium alloys. This Review discusses the specific materials and degradation properties required by all suture materials, summarizes current suture testing protocols and provides a well-grounded direction for the potential future development of biodegradable metal based sutures.

Role of tensile stress and strain in the induction of cell death in experimental vein grafts

Tensile stress and strain are known to induce vascular cell proliferation, a process that is physiologically counterbalanced by cell death. Here we investigate whether tensile stress and strain regulate vascular-cell death by using an end-to-end anastomosed rat vein graft model. In such a model, the circumferential tensile stress in the graft wall was increased by approximately 140 times immediately after surgery compared with that in the venous wall. This change was associated with an increase in the percentage of TUNEL-positive cells at 1, 6, 24, 120, 240, and 720h with two distinct peaks at 1 and 24h (10.1+/-3.5 and 14.4+/-3.2%, respectively) compared with that in control jugular veins (0.4+/-0.5 and 0.5+/-0.5% at 1 and 24h, respectively). When tensile stress and strain in the vein graft wall were reduced by using a biomechanical engineering approach, the rate of cell death was reduced significantly (3.6+/-1.1 and 1.6+/-0.5% at 1 and 24h, respectively). Furthermore, DEVD-CHO, a tetrapeptide aldehyde that inhibits the activity of caspase 3, significantly suppressed this event. These results suggest that a step increase in tensile stress and strain in experimental vein grafts induces rapid cell death, which is possibly mediated by cell death signaling mechanisms.

Rates of in vivo (arterial) and in vitro biocorrosion for pure magnesium.

The development of magnesium-based materials for bioabsorbable stents relies heavily on corrosion testing by immersion in pseudophysiological solutions, where magnesium degrades faster than it does in vivo. The quantitative difference in corrosion kinetics in vitro and in vivo is largely unknown, but, if determined, would help reduce dependence on animal models. In order to create a quantitative in vitro-in vivo correlation based on an accepted measure of corrosion (penetration rate), commercially pure magnesium wires were corroded in vivo in the abdominal aortas of rats for 5-32 days, and in vitro for up to 14 days using Dulbeccos modified eagle medium. Cross-sectioning, scanning electron microscopy, image analysis, a modified penetration rate tailored to degraded wires, and empirical modeling were used to analyze the corroded specimens. In vitro penetration rates were consistently higher than comparable in vivo rates by a factor of 1.2-1.9× (±0.2×). For a sample <20% corroded, an approximate in vitro-in vivo multiplier of 1.3 ± 0.2× was applied, whereas a multiplier of 1.8 ± 0.2× became appropriate when the magnesium specimen was 25-35% degraded.