Directed Assembly of Elastic Fibers via Coacervate Droplet Deposition on Electrospun Templates.

Elastic fibers provide critical elasticity to the arteries, lungs, and other organs. Elastic fiber assembly is a process where soluble tropoelastin is coacervated into liquid droplets, cross-linked, and deposited onto and into microfibrils. While much progress has been made in understanding the biology of this process, questions remain regarding the timing of interactions during assembly. Furthermore, it is unclear to what extent fibrous templates are needed to guide coacervate droplets into the correct architecture. The organization and shaping of coacervate droplets onto a fiber template have never been previously modeled or employed as a strategy for shaping elastin fiber materials. Using an in vitro system consisting of elastin-like polypeptides (ELPs), genipin cross-linker, electrospun polylactic-co-glycolic acid (PLGA) fibers, and tannic acid surface coatings for fibers, we explored ELP coacervation, cross-linking, and deposition onto fiber templates. We demonstrate that integration of coacervate droplets into a fibrous template is primarily influenced by two factors: (1) the balance of coacervation and cross-linking and (2) the surface energy of the fiber templates. The success of this integration affects the mechanical properties of the final fiber network. Our resulting membrane materials exhibit highly tunable morphologies and a range of elastic moduli (0.8-1.6 MPa) comparable to native elastic fibers.

Altered Foreign Body Response at the Posterior Surface Compared to the Anterior Surface of Human Silicone Breast Implants.

Soft implantable devices are crucial to optimizing form and function for many patients. However, periprosthetic capsule fibrosis is one of the major challenges limiting the use of implants. Currently, little is understood about how spatial and temporal factors influence capsule physiology and how the local capsule environment affects the implant structure. In this work, we analyzed breast implant capsule specimens with staining, immunohistochemistry, and real-time polymerase chain reaction to investigate spatiotemporal differences in inflammation and fibrosis. We demonstrated that in comparison to the anterior capsule against the convex surface of breast implants, the posterior capsule against the flat surface of the breast implant displays several features of a dysregulated foreign body reaction including increased capsule thickness, abnormal extracellular remodeling, and infiltration of macrophages. Furthermore, the expression of pro-inflammatory cytokines increased in the posterior capsule across the lifespan of the device, but not in the anterior capsule. We also analyzed the surface oxidation of breast explant samples with XPS analysis. No significant differences in surface oxidation were identified either spatially or temporally. Collectively, our results support spatiotemporal heterogeneity in inflammation and fibrosis within the breast implant capsule. These findings presented here provide a more detailed picture of the complexity of the foreign body reaction surrounding implants destined for human use and could lead to key research avenues and clinical applications to treat periprosthetic fibrosis and improve device longevity.

Structural Integrity of Absorbable Gelatin Sponges for Middle Ear Packing in Otologic Surgery.

OBJECTIVE: This study investigates the physical properties upon immersion of two gelatin sponges commonly used in otologic surgery. BACKGROUND: Absorbable gelatin sponges are often used in middle ear surgery to achieve hemostasis and, perhaps more importantly, to provide a "scaffolding" to support ossicular chain and/or tympanic membrane reconstructions. Their rate of dissolution may therefore affect the success of tympanic membrane closure. METHODS: An in vitro study was conducted to quantify the material changes of two absorbable gelatin sponges, a standard-density sponge and one with fewer collagen cross-linkages (low-density sponge). Volume loss (%) in 0.9% saline, 0.3% ciprofloxacin, and/or 0.1% dexamethasone as single-agent otic drops in a combination formulation was measured at 15-minute intervals for the first hour and at days 1, 3, and 5 postimmersion. Secondary end points included compressibility, porosity under microscopy, and infrared spectroscopy analysis. RESULTS: The low-density sponge immersed in any of the three otic solutions showed a statistically significant greater volume loss at all time points when compared with the standard-density sponge (27.2% ± 5.4% vs. 15.4% ± 6.0% at 15 minutes and 44.8% ± 5.1% vs. 34.6% ± 2.9% at 5 days, p < 0.001). Interestingly, both sponges immersed in normal saline had lost almost half of their original volume after 15 minutes when compared with samples immersed in an otic solution (48.3% ± 4.6% vs. 21.3% ± 8.3%, respectively, p < 0.001). CONCLUSION: The standard-density sponge immersed in an otic solution of ciprofloxacin, dexamethasone, or a combination formulation best maintained its structural integrity. Ancillary in vivo studies are required to assess the hemostatic properties, surgical outcomes, and middle ear synechiae of the above study conditions. LEVEL OF EVIDENCE: Foundational evidence.

Biomimetic Strategy to Enhance Epithelial Cell Viability and Spreading on PEEK Implants.

Polyetheretherketone (PEEK) is a biocompatible material widely used in spinal and craniofacial implants, with potential use in percutaneous implants. However, its inertness prevents it from forming a tight seal with the surrounding soft tissue, which can lead to infections and implant failure. Conversely, the surface chemistry of percutaneous organs (i.e., teeth) helps establish a strong interaction with the epithelial cells of the contacting soft tissues, and hence a tight seal, preventing infection. The seal is created by adsorption of basement membrane (BM) proteins, secreted by epithelial cells, onto the percutaneous organ surfaces. Here, we aim to create a tight seal between PEEK and epithelial tissues by mimicking the surface chemistry of teeth. Our hypothesis is that collagen I, the most abundant tooth protein, enables integration between the epithelial tissue and teeth by promoting adsorption of BM proteins. To test this, we immobilized collagen I via EDC/NHS coupling on a carboxylated PEEK surface modified using diazonium chemistry. We used titanium alloy (Ti-6Al-4V) for comparison, as titanium is the most widely used percutaneous biomaterial. Both collagen-modified PEEK and titanium showed a larger adsorption of key BM proteins (laminin, nidogen, and fibronectin) compared to controls. Keratinocyte epithelial cell viability on collagen-modified PEEK was twice that of control PEEK and ∼1.5 times that of control titanium after 3 days of cell seeding. Both keratinocytes and fibroblasts spread more on collagen-modified PEEK and titanium compared to controls. This work introduces a versatile and biomimetic surface modification technique that may enhance PEEK-epithelial tissue sealing with the potential of extending PEEK applications to percutaneous implants, making it competitive with titanium.

Globule and fiber formation with elastin-like polypeptides: a balance of coacervation and crosslinking.

Elastic fiber assembly is a complex process that requires the coacervation and cross-linking of the protein building block tropoelastin. To date, the order, timing, and interplay of coacervation and crosslinking is not completely understood, despite a great number of advances into understanding the molecular structure and functions of the many proteins involved in elastic fiber assembly. With a simple in vitro model using elastin-like polypeptides and the natural chemical crosslinker genipin, we demonstrate the strong influence of the timing and kinetics of crosslinking reaction on the coacervation, crosslinking extent, and resulting morphology of elastin. We also outline a method for analyzing elastin droplet network formation as a heuristic for measuring the propensity for elastic fiber formation. From this we show that adding crosslinker during peak coacervation dramatically increases the propensity for droplet network formation.

Protein Adsorption on Surfaces Functionalized with COOH Groups Promotes Anti-inflammatory Macrophage Responses.

Implants can induce a foreign body reaction that leads to chronic inflammation and fibrosis in the surrounding tissue. Macrophages help detect the foreign material, play a role in the inflammatory response, and may promote fibrosis instead of the desired tissue regeneration around implants. Implant surface properties impact macrophage responses by changing the nature of the adsorbed protein layer, but conflicting studies highlight the complexity of this relationship. In this study, the effect of surface chemistry on macrophage behavior was investigated with poly(styrene) surfaces containing common functional groups at similar surface densities. The protein layer was characterized to identify the proteins that adsorbed on the surfaces from the medium and the proteins secreted onto the surfaces by adherent macrophages. Of the surface chemistries studied, carboxylic acid (COOH) groups promoted anti-inflammatory responses from unstimulated macrophages and did not exacerbate inflammation upon stimulation. These surfaces also enhanced the adsorption of proteins involved in integrin signaling and promoted the secretion of proteins related to angiogenesis, integrin signaling, and cytokine signaling, which have been previously associated with improved biomaterial integration. Therefore, this study suggests that surface modification with COOH groups may help improve the integration of implants in the body by enhancing anti-inflammatory macrophage responses through altered protein adsorption.

Elastin calcification in in vitro models and its prevention by MGP's N-terminal peptide.

Medial calcification has been associated with diabetes, chronic kidney disease, and genetic disorders like pseudoxanthoma elasticum. Recently, we showed that genetic reduction of arterial elastin content reduces the severity of medial calcification in matrix Gla protein (MGP)-deficient and Eln haploinsufficient Mgp-/-;Eln+/- mice. This study suggests that there might be a direct effect of elastin amount on medial calcification. We studied this using novel in vitro systems, which are based on elastin or elastin-like polypeptides. We first examined the mineral deposition properties of a transfected pigmented epithelial cell line that expresses elastin and other elastic lamina proteins. When grown in inorganic phosphate-supplemented medium, these cells deposited calcium phosphate minerals, which could be prevented by an N'-terminal peptide of MGP (m3pS) carrying phosphorylated serine residues. We next confirmed these findings using a cell-free elastin-like polypeptide (ELP3) scaffold, where the peptide prevented mineral maturation. Overall, this work describes a novel cell culture model for elastocalcinosis and examines the inhibition of mineral deposition by the m3pS peptide in this and a cell-free elastin-based scaffold. Our study provides strong evidence suggesting the critical functional roles of MGP's phosphorylated serine residues in the prevention of elastin calcification and proposes a possible mechanism of their action.

Sobrevivendo no inferno: narrar a vida para fazer algo / Sobrevivir en el inferno: narrar la vida para hacer algo / Surviving in hell: narrating life to get something done

Este trabalho se desenvolve a partir de três eixos. O primeiro é uma breve retomada do que Foucault define como a biopolítica. Em seguida apresentamos e contextualizamos uma ferramenta de análise criada pelo filósofo e cientista social camaronês Achille Mbembe, a necropolítica, que constitui uma importante atualização da crítica social foucaultiana para fenômenos das periferias do capitalismo. De forma mais específica, tomaremos como eixo argumentativo aquilo que alude à articulação entre a biopolítica, a produção da vida e de subjetividades adequadas à forma social capitalista, e a necropolítica, uma política centrada na produção da morte em larga escala. Por fim, a partir dessa análise, tomando como eixo a produção do grupo de rap Racionais MC's, destacaremos duas figuras que emergem das margens das cidades e que são capazes de operar a transmissão: o sobrevivente e a memória. Estas figuras, além de testemunharem a política de morte, podem indicar alguma forma de tratamento possível.

Este trabajo se desarrolla a partir de tres ejes. El primero es una breve revisión de lo que Foucault define como biopolítica. Luego presentamos y contextualizamos una herramienta de análisis creada por el filósofo y científico social camerunés Achille Mbembe, la necro-política, que constituye una actualización importante de la crítica social de Foucault para analizar fenómenos de las periferias del capitalismo. Más específicamente, tomaremos como eje argumentativo lo que alude a la articulación entre la biopolítica, la producción de vida y subjetividades adecuadas a la forma social capitalista, y la necro-política, una política centrada en la producción de muerte a gran escala. Finalmente, con base en este análisis, y tomando como eje la producción del grupo de rap Racionais MC's, destacaremos dos figuras que emergen de los márgenes de las ciudades y que son capaces de operar la transmisión: el sobreviviente y la memoria. Estas figuras, además de dar testimonio de la política de muerte, pueden indicar alguna posible forma de tratamiento.

This work develops from three axes. The first is a brief review of what Foucault defines as biopolitics. Then we present and contextualize an analysis tool created by the Cameroonian philosopher and social scientist Achille Mbembe, necropolitics, which constitutes an important update of Foucault's social criticism to deal with phenomena in the peripheries of capitalism. More specifically, we will take as an argumentative axis what alludes to the articulation between biopolitics, the production of life and subjectivities adequate to the capitalist social form, and necropolitics, a policy centered on the production of death on a large scale. Finally, based on this analysis, and taking the production of the rap group Racionais MC's as an axis, we will highlight two figures that emerge from the margins of cities and that are capable of operating the transmission: the survivor and the memory. These figures, in addition to witnessing the death policy, may indicate some possible form of treatment.

Differences in mineral composition and morphology between men and women in aortic valve calcification.

Aortic valve calcification leads to the deposition of calcium phosphate minerals in the extracellular matrix of the aortic valve leaflets. The mineral deposits can severely narrow the opening of the aortic valve, leading to aortic stenosis. There are no therapies to halt or slow down disease progression and the mechanisms governing aortic valve calcification are still poorly understood. Recently, several studies have shown that for the same aortic stenosis severity, women present significantly lower calcification loads than men. The cause of this sex-related difference is unknown. To understand this difference, we analyzed mineral deposits from surgically excised calcified human aortic valves with different material characterization techniques. We find profound differences in mineral composition and morphology between sexes, which strongly suggest that minerals form slower in women than in men and follow a different mineralization pathway. This finding paves the way for new approaches specifically geared towards men or women in the diagnosis and treatment of aortic valve calcification. STATEMENT OF SIGNIFICANCE: Aortic valve calcification is a health disorder with increasing prevalence and high morbidity and mortality. Currently there is no approved effective treatment; the only available therapeutic option is invasive valve replacement, to which not all patients are suited. The main reason for such lack of treatment options is our lack of understanding of the calcification mechanism. In this study, we show profound differences in mineral composition and morphology between sexes, suggesting that aortic valve calcification follows different mineralization pathways in men and women. These findings pave the way for new approaches specifically geared towards men or women in the diagnosis and treatment of aortic valve calcification.

Effect of the Ionic Concentration of Simulated Body Fluid on the Minerals Formed on Cross-Linked Elastin-Like Polypeptide Membranes.

Deposition of calcium phosphate minerals on the elastin-rich medial layers of arteries can cause severe cardiovascular complications. There are no available treatments for medial calcification, and the mechanism of mineral formation on elastin layers is still unknown. We recently developed an in vitro model of medial calcification using cross-linked elastin-like polypeptide (ELP) membranes immersed in simulated body fluid (SBF). While mineral phase evolution matched that observed in a mouse model of medial calcification, the long incubation required was a practical limitation of this model. Using higher SBF ion concentrations could be a solution to speed up mineral deposition, but its effect on the mineralization process is still not well understood. Here we analyze mineral formation and phase transformation on ELP membranes immersed in high concentration SBF. We show that while mineral deposition is significantly accelerated in these conditions, the chemistry and morphology of the minerals deposited on the ELP membranes and the overall mineralization process are strongly affected. Overall, this work suggests that while the use of low concentration SBF in this in vitro model is more appropriate to study medial calcification associated with the loss of calcification inhibitors, higher SBF ion concentration may be more relevant to study medial calcification in patients with life-threatening diseases such as chronic kidney disease.

Cross-Linked Elastin-like Polypeptide Membranes as a Model for Medial Arterial Calcification.

Calcium phosphate minerals deposit on the elastin-rich medial layers of arteries in the majority of seniors, diabetic, and chronic kidney disease patients, causing severe cardiovascular complications. There is no cure for medial calcification, and the mechanism of mineral formation on elastin layers is unknown. Here we propose cross-linked elastin-like polypeptide membranes as models to study medial calcification. Calcium phosphates deposit first on fibers and filaments and then spread to globular structures present in the membranes. Mineral phase evolution analyzed by near-edge X-ray spectroscopy matches that previously observed in a mouse model of medial calcification, showing that this simple system captures some of the key in vivo findings. This work shows how minerals form and evolve upon nucleation on elastin and provides an in vitro model that can be tuned to study hypotheses related to arterial calcification mechanisms and test drugs to stop or revert mineralization.

Seeing, Targeting and Delivering with Upconverting Nanoparticles.

Efficient control over drug release is critical to increasing drug efficacy and avoiding side effects. An ideal drug delivery system would deliver drugs in the right amount, at the right location and at the right time noninvasively. This can be achieved using light-triggered delivery: light is noninvasive, spatially precise and safe if appropriate wavelengths are chosen. However, the use of light-controlled delivery systems has been limited to areas that are not too deep inside the body because ultraviolet (UV) or visible (Vis) light, the typical wavelengths used for photoreactions, have limited penetration and are toxic to biological tissues. The advent of upconverting nanoparticles (UCNPs) has made it possible to overcome this crucial challenge. UCNPs can convert near-infrared (NIR) radiation, which can penetrate deeper inside the body, to shorter wavelength NIR, Vis and UV radiation. UCNPs have been used as bright, in situ sources of light for on-demand drug release and bioimaging applications. These remote-controlled, NIR-triggered drug delivery systems are especially attractive in applications where a drug is required at a specific location and time such as in anesthetics, postwound healing, cardiothoracic surgery and cancer treatment. In this Perspective, we discuss recent progress and challenges as well as propose potential solutions and future directions, especially with regard to their translation to the clinic.

Raman Spectroscopy for Inverted Papilloma: A Proof-of-Concept Study.

Inverted papillomas are tumors of the sinonasal tract with a propensity to recur. Raman spectroscopy can potentially identify inverted papillomas from other tissue based on biochemical signatures. A pilot study comparing Raman spectroscopy to histopathology for 3 types of sinonasal tissue was performed. Spectral data of biopsies from patients with normal sinonasal mucosa, chronic rhinosinusitis, and inverted papillomas are compared to histopathology using principal component analysis and linear discriminant analysis after data preprocessing. A total of 18 normal, 15 chronic rhinosinusitis, and 18 inverted papilloma specimens were evaluated. The model distinguished normal sinonasal mucosa, chronic rhinosinusitis, and inverted papilloma tissue with an overall accuracy of 90.2% (95% confidence interval, 0.86-0.94). In conclusion, Raman spectroscopy can distinguish inverted papilloma, normal sinonasal mucosa, and chronically rhinosinusitis tissue with acceptable accuracy.

Comparative adsorption profiles of basal lamina proteome and gingival cells onto dental and titanium surfaces.

Titanium (Ti) dental implants are susceptible to bacterial infections and failure due to lack of proper epithelial seal. Epithelial cells establish a strong epithelial seal around natural teeth by the deposition of basal lamina (BL) proteins that adsorb on the tooth surface. This seal can even be re-established onto cementum or dentin following injury or periodontal therapy. However, it is unclear how tooth surfaces promote this cell attachment and protein adsorption. Understanding the interactions between BL proteins and epithelial cells with dentin and Ti will facilitate the development of implant surfaces that promote the formation of an epithelial seal and improve the success of periodontal therapy and wound healing on natural teeth. To study these interactions, we used a surface proteomic approach to decipher the adsorption profile of BL proteins onto Ti and dentin, and correlated these adsorption profiles with in vitro interactions of human gingival fibroblasts and epithelial cells. Results showed that dentin adsorbed higher amounts of key BL proteins, particularly laminin and nidogen-1, and promoted more favorable interactions with epithelial cells than Ti. Next, dentin specimens were deproteinized or partially demineralized to determine if its mineral or protein component was responsible for BL adsorption and cell attachment. Deproteinized (mineral-rich) and partially demineralized (protein-rich) dentin specimens revealed BL proteins (i.e. laminin and nidogen-1) and epithelial cells interact preferentially with dentinal proteins rather than dentin mineral. These findings suggest that, unlike Ti, dentin and, in particular, dentinal proteins have a selective affinity to BL proteins that enhance epithelial cell attachment. STATEMENT OF SIGNIFICANCE: It is remains unclear why natural teeth, unlike titanium dental implants, promote the formation of an epithelial seal that protects them against the external environment. This study used a surface screening approach to analyze the adsorption of proteins produced by epithelial tissues onto tooth-dentin and titanium surfaces, and correlate it with the behaviour of cells. This study shows that tooth-dentin, in particular its proteins, has a higher selective affinity to certain adhesion proteins, and subsequently allows more favourable interactions with epithelial cells than titanium. This knowledge could help in developing new approaches for re-establishing and maintaining the epithelial seal around teeth, and could pave the way for developing implants with surfaces that allow the formation of a true epithelial seal.

Scaffolds for epithelial tissue engineering customized in elastomeric molds.

Restoration of soft tissue defects remains a challenge for surgical reconstruction. In this study, we introduce a new approach to fabricate poly(d,l-lactic acid) (PDLLA) scaffolds with anatomical shapes customized to regenerate three-dimensional soft tissue defects. Highly concentrated polymer/salt mixtures were molded in flexible polyether molds. Microcomputed tomography showed that with this approach it was possible to produce scaffolds with clinically acceptable volume ratio maintenance (>90%). Moreover, this technique allowed us to customize the average pore size and pore interconnectivity of the scaffolds by using variations of salt particle size. In addition, this study demonstrated that with the increasing porosity and/or the decreasing of the average pore size of the PDLLA scaffolds, their mechanical properties decrease and they degrade more slowly. Cell culture results showed that PDLLA scaffolds with an average pore size of 100 µm enhance the viability and proliferation rates of human gingival epithelial cells up to 21 days. The simple method proposed in this article can be extended to fabricate porous scaffolds with customizable anatomical shapes and optimal pore structure for epithelial tissue engineering. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 106B: 880-890, 2018.

Biomaterial surface proteomic signature determines interaction with epithelial cells.

Cells interact with biomaterials indirectly through extracellular matrix (ECM) proteins adsorbed onto their surface. Accordingly, it could be hypothesized that the surface proteomic signature of a biomaterial might determine its interaction with cells. Here, we present a surface proteomic approach to test this hypothesis in the specific case of biomaterial-epithelial cell interactions. In particular, we determined the surface proteomic signature of different biomaterials exposed to the ECM of epithelial cells (basal lamina). We revealed that the biomaterial surface chemistry determines the surface proteomic profile, and subsequently the interaction with epithelial cells. In addition, we found that biomaterials with surface chemistries closer to that of percutaneous tissues, such as aminated PMMA and aminated PDLLA, promoted higher selective adsorption of key basal lamina proteins (laminins, nidogen-1) and subsequently improved their interactions with epithelial cells. These findings suggest that mimicking the surface chemistry of natural percutaneous tissues can improve biomaterial-epithelial integration, and thus provide a rationale for the design of improved biomaterial surfaces for skin regeneration and percutaneous medical devices. STATEMENT OF SIGNIFICANCE: Failure of most biomaterials originates from the inability to predict and control the influence of their surface properties on biological phenomena, particularly protein adsorption, and cellular behaviour, which subsequently results in unfavourable host response. Here, we introduce a surface-proteomic screening approach using a label-free mass spectrometry technique to decipher the adsorption profile of extracellular matrix (ECM) proteins on different biomaterials, and correlate it with cellular behaviour. We demonstrated that the way a biomaterial selectively interacts with specific ECM proteins of a given tissue seems to determine the interactions between the cells of that tissue and biomaterials. Accordingly, this approach can potentially revolutionize the screening methods for investigating the protein-cell-biomaterial interactions and pave the way for deeper understanding of these interactions.

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.

Role of Noncanonical Wnt Signaling Pathway in Human Aortic Valve Calcification.

OBJECTIVE: The mechanisms underlying the pathogenesis of aortic valve calcification remain unclear. With accumulating evidence demonstrating that valve calcification recapitulates bone development, the crucial roles of noncanonical Wnt ligands WNT5a, WNT5b, and WNT11 in osteogenesis make them critical targets in the study of aortic valve calcification. APPROACH AND RESULTS: Using immunohistochemistry, real-time qPCR, Western blotting, and tissue culture, we examined the tissue distribution of WNT5a, WNT5b, and WNT11 in noncalcified and calcified aortic valves and their effects on human aortic valve interstitial cells (HAVICs). Only focal strong immunostaining for WNT5a was seen in and around areas of calcification. Abundant immunostaining for WNT5b and WNT11 was seen in inflammatory cells, fibrosis, and activated myofibroblasts in areas of calcified foci. There was significant correlation between WNT5b and WNT11 overall staining and presence of calcification, lipid score, fibrosis, and microvessels (P<0.05). Real-time qPCR and Western blotting revealed abundant expression of both Wnts in stenotic aortic valves, particularly in bicuspid valves. Incubation of HAVICs from noncalcified valves with the 3 noncanonical Wnts significantly increased cell apoptosis and calcification (P<0.05). Treatment of HAVICs with the mitogen-activated protein kinase-38ß and GSK3ß inhibitors significantly reduced their mineralization (P<0.01). Raman spectroscopy identified the inorganic phosphate deposits as hydroxyapatite and showed a significant increase in hydroxyapatite deposition in HAVICs in response to WNT5a and WNT11 (P<0.05). Similar crystallinity was seen in the deposits found in HAVICs treated with Wnts and in calcified human aortic valves. CONCLUSIONS: These findings suggest a potential role for noncanonical Wnt signaling in the pathogenesis of aortic valve calcification.

Nano graphene oxide-wrapped gold nanostars as ultrasensitive and stable SERS nanoprobes.

We report a facile method to synthesize highly branched gold nanostars wrapped with nano graphene oxide (nGO) sheets with or without the addition of Raman dyes, as nanoprobes with high SERS activity. Both cysteamine and nGO are added to gold nanostars; the positively charged amino groups help self-assembly of nGO flakes around the nanostars. This increases the Raman signal of nGO by 5.3 folds compared to samples in which nGO is in contact with the nanostars but does not wrap them. We also prepare dye-based SERS nanoprobes by sandwiching a typical Raman reporter such as Rhodamine B (RhB), Crystal Violet (CV) and Rhodamine 6G (R6G) between the nanostars and the nGO coating. The Raman signals of RhB are 5.2 times larger when sandwiched between nGO and nanostars than if the molecules are just adsorbed on the nanostar surface, and similar enhancements are observed for the other dyes. In addition to improving SERS efficiency, the wrapping greatly improves the stability of the dye-based nanoprobes, showing a reproducible Raman signal of RhB for over a week in simulated body fluids at 37 °C. High SERS signal, facile fabrication method and excellent stability make these nanoprobes highly promising for SERS-based biosensing and bioimaging applications.

Da rivalidade à responsabilidade: reflexões sobre a justiça restaurativa a partir da psicanálise / From rivalry to accountability: psychoanalytic reflections on restorative justice / De la rivalité à la responsabilité: réflexions sur la justice réparatrice de la psychanalyse / De la rivalidad a la responsabilidad: reflexiones sobre la justicia restaurativa a partir del psicoanálisis

Várias modalidades de intervenção têm incluído as relações horizontais, que apostam na circulação da palavra como condição de inscrição de uma distância capaz de aproximar e produzir um laço com o outro. Neste trabalho, iremos contemplar as funções do outro, abordar a relação com o semelhante, destacando os elementos levantados por Lacan em Hamlet, demonstrando que é para além da rivalidade com o semelhante que entra em jogo a realização do desejo de Hamlet (Lacan, 1959/86). Nossa estratégia será pela apresentação de duas cenas de intervenção junto a adolescentes em conflito com a lei, uma delas em um processo de Justiça Restaurativa. Vamos tirar consequências desta análise no que se refere à possibilidade de condições da horizontalidade nas relações, particularmente nas intervenções junto a adolescentes em conflito com a lei, tendo em vista a responsabilidade como uma construção que se opera no encontro com o outro...

A number of interventions have included horizontal relationships betting on the circulation of the word as a condition for inscribing a distance able to produce a closer bond with the other. This article considers the functions of the other, addresses the relationship with similar others by highlighting the elements raised by Lacan in Hamlet and demonstrating that it is beyond the rivalry with similar others that the fulfilment of Hamlet's desire (Lacan, 1959/86) enters the game. Our strategy will be the presentation of two scenes of intervention with adolescents in conflict with the law, one of whom in a process of restorative justice. We shall take the consequences of this analysis regarding the possibility and conditions of horizontality in the relations, particularly in interventions with adolescents in conflict with the law, aiming at responsibility as a construction which operates in the encounter with the other...

Plusieurs modalités d'intervention incluent les relations horizontales qui de son côté investissent à la circulation de la parole comme condition d'inscription d'une distance capable d'approcher et de produire des liens avec l'autre. Dans ce travail nous allons aborder les fonctions de l'autre, la relation avec le semblable, soulevant les éléments travaillés par Lacan dans Hamlet, pour démontrer que c'est au-delà de la rivalité au semblable qui se joue la réalisation du désir dans Hamlet (Lacan, 1959/86). Dans ce sens, la stratégie se déroulera par la présentation de deux scènes d'intervention auprès des adolescents en conflit avec la loi, dont une est en processus de Justice Réparatrice. Les conséquences de cette analyse seront basées par les possibilités et les conditions d'horizontalité dans les relations, particulièrement dans les interventions auprès des adolescents en conflit avec la loi, étant donné la responsabilité comme une construction qui opère dans la rencontre avec l'autre...

Varios tipos de intervención han incluido las relaciones horizontales que apostan en la circulación de la palabra como condición para la inscripción de uma distancia capaz de producir una unión más estrecha com el otro. En este trabajo, consideramos las funciones del otro, abordamos la relación con otros semejantes destacando los elementos planteados por Lacan en Hamlet, y demonstrando que más allá de la rivalidad con otros semejante entra en el juego el cumplimento del deseo de Hamlet (Lacan, 1959/86 ). Nuestra estrategia será la presentación de la intervención de dos escenas con adolescentes en conflicto con la ley, uno de los quales en un proceso de justicia restaurativa. Tomaremos las consecuencias de este análisis considerando la posibilidad y las condiciones de las relaciones horizontales, sobre todo en las intervenciones con adolescentes en conflicto con la ley, en vista de la responsabilidad como una construcción que opera en el encuentro con el otro...