T cell-loaded injectable chitosan scaffold shows short-term efficacy in localised cancer immunotherapy in mice.

Adoptive cell therapy (ACT) shows success against treatment-resistant cancers, but is limited by the large number of intravenously delivered T cells required and toxicity related to systemic administration. In this work, we hypothesized that localized T cell delivery in an in situ gelling chitosan hydrogel will allow similar treatment efficacy despite delivering fewer cells than systemic intravenous delivery. A rapidly gelling chitosan gel with good mechanical properties was used for this study. Gel biocompatibility and biodegradability were tested over 8 weeks in mice. No adverse effects were observed. The gel elicited a local granulomatous reaction (foreign body reaction), degrading by about 75% volume at 8 weeks. The survival, escape and bioactivity against the tumour cells of encapsulated murine lymphocytes (OT-I) and human Jurkat cells were confirmed in vitro by live/dead assay and flow cytometry. Efficacy was studied using a mouse tumour model where the injected OT-I can specifically recognize and attack ovalbumin (OVA) protein-expressing tumours. The OT-I cell delivery scaffold was compared to untreated controls, OT-I in saline and intravenous systemic treatment with 3-fold more OT-I, observing tumour growth and localization by intravital microscopy and histology. Gel-encapsulated OT-I limited tumour growth significantly up to 11 days after treatment compared to that of untreated mice and mice with longer PBS-suspended OT-I treatment (9 days), but slightly less than that of mice with IV-delivered OT-I treatment (14 days). No significant difference was observed when directly comparing the gel and IV treatments. Although further optimization of the treatment is required, this work shows the feasibility and potential of the chitosan gel for localised OT-I delivery in cancer immunotherapy.

Biomaterials for enhanced immunotherapy.

Cancer immunotherapies have revolutionized the treatment of numerous cancers, with exciting results often superior to conventional treatments, such as surgery and chemotherapy. Despite this success, limitations such as limited treatment persistence and toxic side effects remain to be addressed to further improve treatment efficacy. Biomaterials offer numerous advantages in the concentration, localization and controlled release of drugs, cancer antigens, and immune cells in order to improve the efficacy of these immunotherapies. This review summarizes and highlights the most recent advances in the use of biomaterials for immunotherapies including drug delivery and cancer vaccines, with a particular focus on biomaterials for immune cell delivery.

Chitosan Microbeads Produced by One-Step Scalable Stirred Emulsification: A Promising Process for Cell Therapy Applications.

Cell microencapsulation is a promising approach to improve cell therapy outcomes by protecting injected cells from rapid dispersion and allowing bidirectional diffusion of nutrients, oxygen, and waste that promote cell survival in the target tissues. Here, we describe a simple and scalable emulsification method to encapsulate animal cells in chitosan microbeads using thermosensitive gel formulations without any chemical modification and cross-linker. The process consists of a water-in-oil emulsion where the aqueous phase droplets contain cells (L929 fibroblasts or human mesenchymal stromal cells), chitosan acidic solution and gelling agents (sodium hydrogen carbonate and phosphate buffer or beta-glycerophosphate). The oil temperature is maintained at 37 °C, allowing rapid physical gelation of the microbeads. Alginate beads prepared with the same method were used as a control. Microbeads with a diameter of 300-450 µm were successfully produced. Chitosan and alginate (2% w/v) microbeads presented similar rigidity in compression, but chitosan microbeads endured >80% strain without rupture, while alginate microbeads presented fragile breakage at <50% strain. High cell viability and metabolic activity were observed after up to 7 days in culture for encapsulated cells. Mesenchymal stromal cells encapsulated in chitosan microbeads released higher amounts of the vascular endothelial growth factor after 24 h compared to the cells encapsulated in manually cast macrogels. Moreover, microbeads were injectable through 23G needles without significant deformation or rupture. The emulsion-generated chitosan microbeads are a promising delivery vehicle for therapeutic cells because of their cytocompatibility, biodegradation, mechanical strength, and injectability. Clinical-scale encapsulation of therapeutic cells such as mesenchymal stromal cells in chitosan microbeads can readily be achieved using this simple and scalable emulsion-based process.

Chitosan-doxycycline hydrogel: An MMP inhibitor/sclerosing embolizing agent as a new approach to endoleak prevention and treatment after endovascular aneurysm repair.

The success of endovascular repair of abdominal aortic aneurysms remains limited due to the development of endoleaks. Sac embolization has been proposed to manage endoleaks, but current embolizing materials are associated with frequent recurrence. An injectable agent that combines vascular occlusion and sclerosing properties has demonstrated promise for the treatment of endoleaks. Moreover, the inhibition of aneurysmal wall degradation via matrix metalloproteinases (MMPs) may further prevent aneurysm progression. Thus, an embolization agent that promotes occlusion, MMP inhibition and endothelial ablation was hypothesized to provide a multi-faceted approach for endoleak treatment. In this study, an injectable, occlusive chitosan (CH) hydrogel containing doxycycline (DOX)-a sclerosant and MMP inhibitor-was developed. Several CH-DOX hydrogel formulations were characterized for their mechanical and sclerosing properties, injectability, DOX release rate, and MMP inhibition. An optimized formulation was assessed for its short-term ability to occlude blood vessels in vivo. All formulations were injectable and gelled rapidly at body temperature. Only hydrogels prepared with 0.075M sodium bicarbonate and 0.08M phosphate buffer as the gelling agent presented sufficient mechanical properties to immediately impede physiological flow. DOX release from this gel was in a two-stage pattern: a burst release followed by a slow continuous release. Released DOX was bioactive and able to inhibit MMP-2 activity in human glioblastoma cells. Preliminary in vivo testing in pig renal arteries showed immediate and delayed embolization success of 96% and 86%, respectively. Altogether, CH-DOX hydrogels appear to be promising new multifunctional embolic agents for the treatment of endoleaks. STATEMENT OF SIGNIFICANCE: An injectable embolizing chitosan hydrogel releasing doxycycline (DOX) was developed as the first multi-faceted approach for the occlusion of blood vessels. It combines occlusive properties with DOX sclerosing and MMP inhibition properties, respectively known to prevent recanalization process and to counteract the underlying pathophysiology of vessel wall degradation and aneurysm progression. After drug release, the biocompatible scaffold can be invaded by cells and slowly degrade. Local DOX delivery requires lower drug amount and decreases risks of side effects compared to systemic administration. This new gel could be used for the prevention or treatment of endoleaks after endovascular aneurysm repair, but also for the embolization of other blood vessels such as venous or vascular malformations.

Bioavailability of immobilized epidermal growth factor: Covalent versus noncovalent grafting.

In an effort to rationalize and optimize an antiapoptotic coating combining chondroitin sulfate (CS) and epidermal growth factor (EGF) for vascular applications, the authors here report the comparison of two grafting strategies aiming to display EGF in an oriented fashion on CS. For that purpose, the authors produced, purified, and characterized a chimeric protein corresponding to EGF that was N-terminally fused to a cysteine and a coil peptide. The chimera was covalently immobilized via its free thiol group or captured via coiled-coil interactions at the surface of a biosensor or on a chondroitin sulfate coating in multiwell plates, mimicking the coating that was previously developed by them for stent-graft surfaces. The interactions of grafted EGF with the soluble domain of its receptor or the impact of grafted EGF upon vascular smooth muscle survival in proapoptotic conditions indicated that the coiled-coil based tethering was the best approach to display EGF. These results, combined to direct enzyme-linked immunosorbent assay measurements, indicated that the coiled-coil tethering approach allowed increasing the amount of bioavailable EGF when compared to covalent coupling, rather than the total amount of grafted EGF, while using much lower concentrations of tagged EGF during incubation.

Optimization of Injectable Thermosensitive Scaffolds with Enhanced Mechanical Properties for Cell Therapy.

Strong injectable chitosan thermosensitive hydrogels can be created, without chemical modification, by combining sodium hydrogen carbonate with another weak base, namely, beta-glycerophosphate (BGP) or phosphate buffer (PB). Here the influence of gelling agent concentration on the mechanical properties, gelation kinetics, osmolality, swelling, and compatibility for cell encapsulation, is studied in order to find the most optimal formulations and demonstrate their potential for cell therapy and tissue engineering. The new formulations present up to a 50-fold increase of the Young's modulus after gelation compared with conventional chitosan-BGP hydrogels, while reducing the ionic strength to the level of iso-osmolality. Increasing PB concentration accelerates gelation but reduces the mechanical properties. Increasing BGP also has this effect, but to a lesser extent. Cells can be easily encapsulated by mixing the cell suspension within the hydrogel solution at room temperature, prior to rapid gelation at body temperature. After encapsulation, L929 mouse fibroblasts are homogeneously distributed within scaffolds and present a strongly increased viability and growth, when compared with conventional chitosan-BGP hydrogels. Two particularly promising formulations are evaluated with human mesenchymal stem cells. Their viability and metabolic activity are maintained over 7 d in vitro.

Abdominal aortic aneurysm follow-up by shear wave elasticity imaging after endovascular repair in a canine model.

OBJECTIVES: To investigate if shear wave imaging (SWI) can detect endoleaks and characterize thrombus organization in abdominal aortic aneurysms (AAAs) after endovascular aneurysm repair. METHODS: Stent grafts (SGs) were implanted in 18 dogs after surgical creation of type I endoleaks (four AAAs), type II endoleaks (13 AAAs) and no endoleaks (one AAA). Color flow Doppler ultrasonography (DUS) and SWI were performed before SG implantation (baseline), on days 7, 30 and 90 after SG implantation, and on the day of the sacrifice (day 180). Angiography, CT scans and macroscopic tissue sections obtained on day 180 were evaluated for the presence, size and type of endoleaks, and thrombi were characterized as fresh or organized. Endoleak areas in aneurysm sacs were identified on SWI by two readers and compared with their appearance on DUS, CT scans and macroscopic examination. Elasticity moduli were calculated in different regions (endoleaks, and fresh and organized thrombi). RESULTS: All 17 endoleaks (100 %) were identified by reader 1, whereas 16 of 17 (94 %) were detected by reader 2. Elasticity moduli in endoleaks, and in areas of organized thrombi and fresh thrombi were 0.2 ± 0.4, 90.0 ± 48.2 and 13.6 ± 4.5 kPa, respectively (P < 0.001 between groups). SWI detected endoleaks while DUS (three endoleaks) and CT (one endoleak) did not. CONCLUSIONS: SWI has the potential to detect endoleaks and evaluate thrombus organization based on the measurement of elasticity. KEY POINTS: • SWI has the potential to detect endoleaks in post-EVAR follow-up. • SWI has the potential to characterize thrombus organization in post-EVAR follow-up. • SWI may be combined with DUS in post-EVAR surveillance of endoleak.

Mucoadhesive chitosan hydrogels as rectal drug delivery vessels to treat ulcerative colitis.

Mucoadhesive drug delivery systems stick to mucosal tissues and prolong the local retention time of drugs. Since the colon is covered by a mucosal layer, mucoadhesive rectal formulations may improve treatment of such diseases as hypertension or colon cancer. Ulcerative colitis (UC) is an inflammatory bowel disease characterized by chronic inflammation of the colonic mucosa. It is commonly treated with sulfasalazine (SSZ), which is metabolized by the intestinal flora into the therapeutic 5-aminosalicylic acid (5-ASA) and a toxic by-product sulfapyridine (SP). SSZ can be administered orally or rectally. The latter route avoids unintended absorption of the drug or its degradation products in the upper gastrointestinal tract, but often fails due to limited retention time. Here, we propose a mucoadhesive hydrogel to improve the efficacy of rectal SSZ administration. The gel is made of catechol modified-chitosan (Cat-CS) crosslinked by genipin. After loading the gel with SSZ, we evaluated its efficacy in a mouse model of UC. Compared to oral SSZ treatment, rectal SSZ/Cat-CS delivery was more therapeutic, showed equivalent histological scores, and induced a lower plasma concentration of the potentially toxic SP by-product. These results show SSZ/Cat-CS rectal hydrogels are more effective and safer formulations for UC treatment than oral SSZ. STATEMENT OF SIGNIFICANCE: Ulcerative colitis affects the colon by causing chronic inflammation on the mucosa. One of the most common drugs to treat mild to moderate UC is sulfasalazine, which can be administrated both orally and rectally. Rectal formulations are preferable, since their therapeutic effect happens topically, and they prevent side effects related to absorption of the drug in the small intestine. However, the efficacy of rectal sulfasalazine formulations is decreased by their limited colon residence time. Here we propose a chitosan-catechol mucoadhesive gel that allows delivering sulfasalazine more effectively and safely than oral administration. Our results bring new insights into the field of mussel-inspired catechol hydrogels, showing their potential as drug delivery systems to treat a widespread disease such as ulcerative colitis.

Experimental validation of more realistic computer models for stent-graft repair of abdominal aortic aneurysms, including pre-load assessment.

Although the endovascular repair of abdominal aortic aneurysms is a less invasive alternative than classic open surgery, complications such as endoleak and kinking still need to be addressed. Numerical simulation of endovascular repair is becoming a valuable tool in stent-graft (SG) optimization, patient selection and surgical planning. The experimental and numerical forces required to produce SG deformations were compared in a range of in vivo conditions in the present study. The deformation modes investigated were: bending as well as axial, transversal and radial compressions. In particular, an original method was developed to efficiently account for radial pre-load because of the pre-compression of stents to match the graft dimensions during manufacturing. This is important in order to compute the radial force exerted on the vessel after deployment more accurately. Variations of displacement between the experimental and numerical results ranged from 1.39% for simple leg bending to 5.93% for three-point body bending. Finally, radial pre-load was modeled by increasing Young's modulus of each stent. On average, it was found that Young's modulus had to be augmented by a factor of 2. Copyright © 2016 John Wiley & Sons, Ltd.

Controlled co-immobilization of EGF and VEGF to optimize vascular cell survival.

Growth factors (GFs) are potent signaling molecules that act in a coordinated manner in physiological processes such as tissue healing or angiogenesis. Co-immobilizing GFs on materials while preserving their bioactivity still represents a major challenge in the field of tissue regeneration and bioactive implants. In this study, we explore the potential of an oriented immobilization technique based on two high affinity peptides, namely the Ecoil and Kcoil, to allow for the simultaneous capture of the epidermal growth factor (EGF) and the vascular endothelial growth factor (VEGF) on a chondroitin sulfate coating. This glycosaminoglycan layer was selected as it promotes cell adhesion but reduces non-specific adsorption of plasma proteins. We demonstrate here that both Ecoil-tagged GFs can be successfully immobilized on chondroitin sulfate surfaces that had been pre-decorated with the Kcoil peptide. As shown by direct ELISA, changing the incubation concentration of the various GFs enabled to control their grafted amount. Moreover, cell survival studies with endothelial and smooth muscle cells confirmed that our oriented tethering strategy preserved GF bioactivity. Of salient interest, co-immobilizing EGF and VEGF led to better cell survival compared to each GF captured alone, suggesting a synergistic effect of these GFs. Altogether, these results demonstrate the potential of coiled-coil oriented GF tethering for the co-immobilization of macromolecules; it thus open the way to the generation of biomaterials surfaces with fine-tuned biological properties. STATEMENT OF SIGNIFICANCE: Growth factors are potent signaling molecules that act in a coordinated manner in physiological processes such as tissue healing or angiogenesis. Controlled coimmobilization of growth factors on biomaterials while preserving their bioactivity represents a major challenge in the field of tissue regeneration and bioactive implants. This study demonstrates the potential of an oriented immobilization technique based on two high affinity peptides to allow for the simultaneous capture of epidermal growth factor (EGF) and vascular endothelial growth factor (VEGF). Our system allowed an efficient control on growth factor immobilization by adjusting the incubation concentrations of EGF and VEGF. Of salient interest, co-immobilizing of specific ratios of EGF and VEGF demonstrated a synergistic effect on cell survival compared to each GF captured alone.

Chitosan thermogels for local expansion and delivery of tumor-specific T lymphocytes towards enhanced cancer immunotherapies.

The success of promising anti-cancer adoptive cell therapies relies on the abilities of the perfused CD8(+) T lymphocytes to gain access to and persist within the tumor microenvironment to carry out their cytotoxic functions. We propose a new method for their local delivery as a living concentrate, which may not only reduce the numbers of cells required for treatment but also enhance their site-specific mobilization. Using combinations of sodium hydrogen carbonate and phosphate buffer as gelling agents, novel injectable chitosan-based biocompatible thermogels (CTGels) having excellent mechanical properties and cytocompatibility have been developed. Three thermogel formulations with acceptable physicochemical properties, such as physiological pH and osmolality, macroporosity, and gelation rates were compared. The CTGel2 formulation outperformed the others by providing an environment suitable for the encapsulation of viable CD8(+) T lymphocytes, supporting their proliferation and gradual release. In addition, the encapsulated T cell phenotypes were influenced by surrounding conditions and by tumor cells, while maintaining their capacity to kill tumor cells. This strongly suggests that cells encapsulated in this formulation retain their anti-cancer functions, and that this locally injectable hydrogel may be further developed to complement a wide variety of existing immunotherapies.

Endovascular Repair of Abdominal Aortic Aneurysm: Follow-up with Noninvasive Vascular Elastography in a Canine Model.

PURPOSE: To assess the ability of noninvasive vascular elastography (NIVE) to help characterize endoleaks and thrombus organization in a canine model of abdominal aortic aneurysm after endovascular aneurysm repair with stent-grafts, in comparison with computed tomography (CT) and pathologic examination findings. MATERIALS AND METHODS: All protocols were approved by the Animal Care Committee in accordance with the guidelines of the Canadian Council of Animal Care. Stent-grafts were implanted in a group of 18 dogs with aneurysms created in the abdominal aorta. Type I endoleak was created in four aneurysms; type II endoleak, in 13 aneurysms; and no endoleak, in one aneurysm. Doppler ultrasonography and NIVE examinations were performed at baseline and at 1-week, 1-month, 3-month, and 6-month follow-up. Angiography, CT, and macroscopic tissue examination were performed at sacrifice. Strain values were computed by using the Lagrangian speckle model estimator. Areas of endoleak, solid organized thrombus, and fresh thrombus were identified and segmented by comparing the results of CT and macroscopic tissue examination. Strain values were compared by using the Wilcoxon rank-sum and Kruskal-Wallis tests. RESULTS: All stent-grafts were successfully deployed, and endoleaks were clearly depicted in the last follow-up elastography examinations. Maximal axial strains over consecutive heart cycles in endoleak, organized thrombus, and fresh thrombus areas were 0.78% ± 0.22, 0.23% ± 0.02, 0.10% ± 0.04, respectively. Strain values were significantly different between endoleak and organized or fresh thrombus areas (P < .000) and between organized and fresh thrombus areas (P < .0002). No correlation was found between strain values and type of endoleak, sac pressure, endoleak size, and aneurysm size. CONCLUSION: NIVE may be able to help characterize endoleak and thrombus organization, regardless of the size, pressure, and type of endoleak.

Energetics of Molecular Excitation, Fragmentation, and Polymerization in a Dielectric Barrier Discharge with Argon Carrier Gas.

We report experiments at atmospheric pressure (AP) using a dielectric barrier discharge (DBD) reactor designed for plasma polymerization (PP) with "monomers" at ‰ concentrations in ca.10 standard liters per minute of argon (Ar) carrier gas. We have perfected a method for measuring Eg, the energy dissipated per cycle of the applied a.c. high voltage, Va(f), but the focus here is on ΔEg, the energy difference with and without a flow, Fd, of monomer in the Ar flow, with the plasma being sustained at Va(f) = 2.8 kVrms, f = 20 kHz. From ΔEg and Fd, we derive a characteristic energy per molecule, Em (in eV), and investigate plots of Em versus Fd and 1/Fd for three model "monomers": formic, acetic, and acrylic acid. These data, along with those for lighter or heavier organic compounds, reveal novel information about energy absorption from the plasma and ensuing polymerization reactions.

A literature review of the numerical analysis of abdominal aortic aneurysms treated with endovascular stent grafts.

The purpose of this paper is to present the basic principles and relevant advances in the computational modeling of abdominal aortic aneurysms and endovascular aneurysm repair, providing the community with up-to-date state of the art in terms of numerical analysis and biomechanics. Frameworks describing the mechanical behavior of the aortic wall already exist. However, intraluminal thrombus nonhomogeneous structure and porosity still need to be well characterized. Also, although the morphology and mechanical properties of calcifications have been investigated, their effects on wall stresses remain controversial. Computational fluid dynamics usually assumes a rigid artery wall, whereas fluid-structure interaction accounts for artery compliance but is still challenging since arteries and blood have similar densities. We discuss alternatives to fluid-structure interaction based on dynamic medical images that address patient-specific hemodynamics and geometries. We describe initial stresses, elastic boundary conditions, and statistical strength for rupture risk assessment. Special emphasis is accorded to workflow development, from the conversion of medical images into finite element models, to the simulation of catheter-aorta interactions and stent-graft deployment. Our purpose is also to elaborate the key ingredients leading to virtual stenting and endovascular repair planning that could improve the procedure and stent-grafts.

Stem cells, nitrogen-rich plasma-polymerized culture surfaces, and type X collagen suppression.

Mesenchymal stem cells (MSCs) are multipotent cells that can differentiate into chondrocytes, osteoblasts, myocytes, adipocytes, and a variety of other cell types. Several studies have been directed toward using MSCs from patients with osteoarthritis (OA) for cartilage repair, not only because these are the ones that will require a source of autologous stem cells if biological repair of cartilage lesions is to be a therapeutic option, but also to further an understanding of stem cell differentiation. Previous studies have shown that a major drawback of current cartilage and intervertebral disc tissue repair is that human MSCs from OA patients express type X collagen (COL X). COL X, a marker of late-stage chondrocyte hypertrophy, is implicated in endochondral ossification. However, those studies also revealed that a novel plasma-polymerized thin film material, named nitrogen-rich plasma-polymerized ethylene (PPE:N), was able to inhibit COL X expression in committed MSCs. The specific aim of this present study was to determine if the suppression of COL X by PPE:N is maintained when MSCs are transferred to pellet cultures in serum-free media. Our results confirmed the potential of two different types of PPE:N surfaces (low-pressure-PPE:N [L-PPE:N] and high-pressure-PPE:N [H-PPE:N]) in suppressing COL X expression, more so on the latter. Interestingly, when MSCs were transferred to pellet cultures, the expression level of COL X was further decreased by preincubation on H-PPE:N, suggesting that these kinds of coatings show promise for tissue engineering of cartilage and disc tissues. Further studies are needed to assess the relative importance of surface-chemistry versus surface-morphology in the mechanism of COL X suppression.

Noninvasive vascular ultrasound elastography applied to the characterization of experimental aneurysms and follow-up after endovascular repair.

Experimental and simulation studies were conducted to noninvasively characterize abdominal aneurysms with ultrasound (US) elastography before and after endovascular treatment. Twenty three dogs having bilateral aneurysms surgically created on iliac arteries with venous patches were investigated. In a first set of experiments, the feasibility of elastography to differentiate vascular wall elastic properties between the aneurismal neck (healthy region) and the venous patch (pathological region) was evaluated on six dogs. Lower strain values were found in venous patches (p < 0.001). In a second set of experiments, 17 dogs having endovascular repair (EVAR) by stent graft (SG) insertion were examined three months after SG implantation. Angiography, color Doppler US, examination of macroscopic sections and US elastography were used. The value of elastography was validated with the following end points by considering a solid thrombus of a healed aneurysm as a structure with small deformations and a soft thrombus associated with endoleaks as a more deformable tissue: (1) the correlation between the size of healed organized thrombi estimated by elastography and by macroscopic examinations; (2) the correlation between the strain amplitude measured within vessel wall elastograms and the leak size; and (3) agreement on the presence and size of endoleaks as determined by elastography and by combined reference imaging modalities (angiography + Doppler US). Mean surfaces of solid thrombi estimated with elastography were found correlated with those measured on macroscopic sections (r = 0.88, p < 0.001). Quantitative strain values measured within the vessel wall were poorly linked with the leak size (r = 0.12, p = 0.5). However, the qualitative evaluation of leak size in the aneurismal sac was very good, with a Kappa agreement coefficient of 0.79 between elastography and combined reference imaging modalities. In summary, complementing B-scan and color Doppler, noninvasive US elastography was found to be potentially a relevant tool for aneurismal follow-up after EVAR, provided it allows geometrical and mechanical characterizations of the solid thrombus within the aneurismal sac. This elasticity imaging technique might help detecting potential complications during follows-up subsequent to EVAR.

Role of the endothelial lining in endoleak formation and persistence after endovascular repair of aneurysm.

PURPOSE: To evaluate whether ablation of the endothelial lining of an aneurysm can prevent endoleak persistence after endovascular aneurysm repair (EVAR). MATERIALS AND METHODS: Bilateral aneurysms were constructed in common iliac arteries in three different experimental groups. In group 1 (six dogs), 12 aneurysms without collateral outflow were created, the endothelial layer was removed on one side, and stent-grafts were implanted immediately after surgery with endovascular creation of a type I endoleak. In group 2 (six dogs), the same experiment was performed in aneurysms with collateral outflow. In group 3 (three dogs), six aneurysms with collateral outflow were denuded, but stent-grafts were implanted 3 months later. Follow-up imaging was performed with Doppler ultrasonography (US) and angiography until animal sacrifice 3 months after EVAR. Leak persistence and healing were also evaluated with macroscopic and histopathologic examination. RESULTS: Type I endoleaks persisted in all endothelialized group 1 aneurysms (six of six), but in none that were denuded before stent-graft implantation (P = .03). The ratio between the maximum diameter of the aneurysm measured before sacrifice and at baseline was significantly lower in denuded aneurysms than in aneurysms with an intact endothelial layer (74% vs 92%, P = .003). Endoleaks were observed in all aneurysms of groups 2 (denuded and intact endothelium) and 3. All endoleak areas were surrounded by endothelial lining. CONCLUSIONS: In this animal model of EVAR, ablation of the endothelial lining promotes long-term endoleak thrombosis after EVAR. The presence of collateral flow can promote re-endothelialization and endoleak persistence.

The effects of stenting and endothelial denudation on aneurysm and branch occlusion in experimental aneurysm models.

BACKGROUND: Stents are increasingly used in the endovascular treatment of intracranial aneurysms. We studied the effects of stenting and endothelial denudation on aneurysm and branch vessel occlusion. METHODS: Bilateral lingual bifurcation venous pouch aneurysms were created in eight dogs, surgically scraping the aneurysmal endothelial lining on one side. Both arteries were immediately stented using balloon-expandable stents. In four other dogs, a wide-neck carotid bifurcation aneurysm was created, with the vein pouch denuded or not (n=2 each), followed by immediate stenting. Results were compared using angiography and pathology at 10 days (n=2), 10 (n=8), and 20 weeks (n=2). Branch occlusion between initial and final angiograms was recorded. Pathological evaluation of aneurysms was studied, with attention to neointima formation at the aneurysm ostium and around branch vessel origins. RESULTS: All stented and denuded lingual aneurysms were obliterated compared with two of eight lingual aneurysms that were stented alone (P=.007). None of the carotid bifurcation aneurysms became obliterated (0/4), but denuded aneurysms showed partial thrombosis (2/2). Of 68 total stent-covered branches, 5 (7%) were occluded and 17 (27%) had altered angiographic flow. CONCLUSIONS: Stenting led to suboptimal results in the presence of an intact endothelial layer. Endothelial denudation can promote aneurysm occlusion when combined with stenting.

Suppression of genes related to hypertrophy and osteogenesis in committed human mesenchymal stem cells cultured on novel nitrogen-rich plasma polymer coatings.

Mesenchymal stem cells (MSCs) are pluripotent progenitor cells with the ability to generate cartilage, bone, muscle, tendon, ligament, and fat. However, recent evidence indicates that a major drawback of current cartilage- and intervertebral disc-tissue engineering is that human MSCs isolated from some arthritic patients (a clinically relevant source of stem cells) express type X collagen (a marker of chondrocyte hypertrophy associated with endochondral ossification) and osteogenic markers. Some studies have attempted to use growth factors to inhibit type X collagen expression, but none has addressed the possible effect of the chemical composition of the substratum on chondrocyte hypertrophy and osteogenesis. Here, we examine the growth and differentiation potential of human MSCs cultured on nitrogen (N)-rich plasma polymer layers (N-doped plasma-polymerized ethylene, containing up to 36% nitrogen; PPE:N). We show that PPE:N almost completely suppresses the expression not only of type X collagen, but also of osteogenic marker genes such as alkaline phosphatase, bone sialoprotein, and osteocalcin. In contrast, neither aggrecan nor type I collagen expression were significantly affected. These results indicate that PPE:N coatings may be suitable surfaces for inducing MSCs to a chondrocyte or disc-like phenotype for tissue engineering of cartilage or intervertebral discs, in which hypertrophy and osteogenesis are suppressed.

Effect of radioactivity on stent-graft incorporation after endovascular treatment of aneurysms: An animal study.

Poor stent-graft (SG) incorporation into the vessel wall, following endovascular repair of abdominal aortic aneurysms (EVAR), can lead to endoleaks and SG migration. Low-dose radiation can prevent aneurysm recurrence after coil embolization, and has been associated with a "paradoxical" increase in neointima formation after stenting in a few studies. It was hypothesized that in situ beta radiation emitted from SG could improve its incorporation by preventing the persistence of circulating channels between the implant and the vessel wall and increasing neointima formation around the SG. Phosphorus 32 ((32)P, 200 or 400 kBq per SG (n = 6 each)) was ion implanted on the external surface of balloon-expandable SGs. Twelve radioactive and six non-radioactive SGs were deployed in iliac arteries of nine Mongrel dogs. Neointima formation inside the graft and the persistence of circulating flow through an artificial groove created during the endovascular procedure were assessed by follow-up imaging and by blinded, computerized histomorphometric analysis after animal sacrifice at 3 months. Occlusion occurred in four radioactive SGs. A lesser number of patent grooves was observed along high-activity SGs than along control SGs (1/3 versus 4/4). No difference in neointima formation was observed in radioactive and non-radioactive SGs. Alteration of external graft surface was observed after ion implantation. Ion implantation of (32)P on SGs does not seem to be a viable strategy to improve incorporation and prevent type-I endoleak after EVAR.