RESUMEN
Desmoplasia in breast cancer leads to heterogeneity in physical properties of the tissue, resulting in disparities in drug delivery and treatment efficacy among patients, thus contributing to high disease mortality. Personalized in vitro breast cancer models hold great promise for high-throughput testing of therapeutic strategies to normalize the aberrant microenvironment in a patient-specific manner. Here, tumoroids assembled from breast cancer cell lines (MCF7, SKBR3, and MDA-MB-468) and patient-derived breast tumor cells (TCs) cultured in microphysiological systems including perfusable microvasculature reproduce key aspects of stromal and vascular dysfunction causing impaired drug delivery. Models containing SKBR3 and MDA-MB-468 tumoroids show higher stromal hyaluronic acid (HA) deposition, vascular permeability, interstitial fluid pressure (IFP), and degradation of vascular HA relative to models containing MCF7 tumoroids or models without tumoroids. Interleukin 8 (IL8) secretion is found responsible for vascular dysfunction and loss of vascular HA. Interventions targeting IL8 or stromal HA normalize vascular permeability, perfusion, and IFP, and ultimately enhance drug delivery and TC death in response to perfusion with trastuzumab and cetuximab. Similar responses are observed in patient-derived models. These microphysiological systems can thus be personalized by using patient-derived cells and can be applied to discover new molecular therapies for the normalization of the tumor microenvironment.
RESUMEN
Endothelial programmed death-ligand 1 (PD-L1) expression is higher in tumors than in normal tissues. Also, tumoral vasculatures tend to be leakier than normal vessels leading to a higher trans-endothelial or transmural fluid flow. However, it is not clear whether such elevated transmural flow can control endothelial PD-L1 expression. Here, a new microfluidic device is developed to investigate the relationship between transmural flow and PD-L1 expression in microvascular networks (MVNs). After treating the MVNs with transmural flow for 24 h, the expression of PD-L1 in endothelial cells is upregulated. Additionally, CD8 T cell activation by phytohemagglutinin (PHA) is suppressed when cultured in the MVNs pre-conditioned with transmural flow. Moreover, transmural flow is able to further increase PD-L1 expression in the vessels formed in the tumor microenvironment. Finally, by utilizing blocking antibodies and knock-out assays, it is found that transmural flow-driven PD-L1 upregulation is controlled by integrin αVß3. Overall, this study provides a new biophysical explanation for high PD-L1 expression in tumoral vasculatures.
Asunto(s)
Antígeno B7-H1 , Microvasos , Regulación hacia Arriba , Antígeno B7-H1/metabolismo , Antígeno B7-H1/genética , Humanos , Microvasos/metabolismo , Microambiente Tumoral , Ratones , Animales , Células Endoteliales/metabolismoRESUMEN
Amphotericin B (AmB) is the gold standard for antifungal drugs. However, AmB systemic administration is restricted because of its side effects. Here, we report AmB loaded in natural rubber latex (NRL), a sustained delivery system with low toxicity, which stimulates angiogenesis, cell adhesion and accelerates wound healing. Physicochemical characterizations showed that AmB did not bind chemically to the polymeric matrix. Electronic and topographical images showed small crystalline aggregates from AmB crystals on the polymer surface. About 56.6% of AmB was released by the NRL in 120 h. However, 33.6% of this antifungal was delivered in the first 24 h due to the presence of AmB on the polymer surface. The biomaterial's excellent hemo- and cytocompatibility with erythrocytes and human dermal fibroblasts (HDF) confirmed its safety for dermal wound application. Antifungal assay against Candida albicans showed that AmB-NRL presented a dose-dependent behavior with an inhibition halo of 30.0 ± 1.0 mm. Galleria mellonella was employed as an in vivo model for C. albicans infection. Survival rates of 60% were observed following the injection of AmB (0.5 mg.mL-1) in G. mellonella larvae infected by C. albicans. Likewise, AmB-NRL (0.5 mg.mL-1) presented survival rates of 40%, inferring antifungal activity against fungus. Thus, NRL adequately acts as an AmB-sustained release matrix, which is an exciting approach, since this antifungal is toxic at high concentrations. Our findings suggest that AmB-NRL is an efficient, safe, and reasonably priced ($0.15) dressing for the treatment of cutaneous fungal infections.
Asunto(s)
Candidiasis , Infección de Heridas , Humanos , Anfotericina B , Antifúngicos/química , Vendajes , Candida albicans , Candidiasis/tratamiento farmacológico , Látex , Pruebas de Sensibilidad Microbiana , Infección de Heridas/tratamiento farmacológicoRESUMEN
Tumor-associated inflammation drives cancer progression and therapy resistance, with the infiltration of monocyte-derived tumor-associated macrophages (TAMs) associated with poor prognosis in diverse cancers. Targeting TAMs holds potential against solid tumors, but effective immunotherapies require testing on immunocompetent human models prior to clinical trials. Here, we develop an in vitro model of microvascular networks that incorporates tumor spheroids or patient tissues. By perfusing the vasculature with human monocytes, we investigate monocyte trafficking into the tumor and evaluate immunotherapies targeting the human tumor microenvironment. Our findings demonstrate that macrophages in vascularized breast and lung tumor models can enhance monocyte recruitment via TAM-produced CCL7 and CCL2, mediated by CSF-1R. Additionally, we assess a novel multispecific antibody targeting CCR2, CSF-1R, and neutralizing TGF-ß, referred to as CSF1R/CCR2/TGF-ß Ab, on monocytes and macrophages using our 3D models. This antibody repolarizes TAMs towards an anti-tumoral M1-like phenotype, reduces monocyte chemoattractant protein secretion, and effectively blocks monocyte migration. Finally, we show that the CSF1R/CCR2/TGF-ß Ab inhibits monocyte recruitment in patient-specific vascularized tumor models. Overall, this vascularized tumor model offers valuable insights into monocyte recruitment and enables functional testing of innovative therapeutic antibodies targeting TAMs in the tumor microenvironment (TME).
RESUMEN
Trans-endothelial electrical resistance (TEER) is one of the most widely used indicators to quantify the barrier integrity of endothelial layers. Over the last decade, the integration of TEER sensors into organ-on-a-chip (OOC) platforms has gained increasing interest for its efficient and effective measurement of TEER in OOCs. To date, microfabricated electrodes or direct insertion of wires has been used to integrate TEER sensors into OOCs, with each method having advantages and disadvantages. In this study, we developed a TEER-SPE chip consisting of carbon-based screen-printed electrodes (SPEs) embedded in a poly(methyl methacrylate) (PMMA)-based multi-layered microfluidic device with a porous poly(ethylene terephthalate) membrane in-between. As proof of concept, we demonstrated the successful cultures of hCMEC/D3 cells and the formation of confluent monolayers in the TEER-SPE chip and obtained TEER measurements for 4 days. Additionally, the TEER-SPE chip could detect changes in the barrier integrity due to shear stress or an inflammatory cytokine (i.e., tumor necrosis factor-α). The novel approach enables a low-cost and facile fabrication of carbon-based SPEs on PMMA substrates and the subsequent assembly of PMMA layers for rapid prototyping. Being cost-effective and cleanroom-free, our method lowers the existing logistical and technical barriers presenting itself as another step forward to the broader adoption of OOCs with TEER measurement capability.
Asunto(s)
Sistemas Microfisiológicos , Polimetil Metacrilato , Impedancia Eléctrica , Carbono , ElectrodosRESUMEN
Films and coatings manufactured with bio-based renewable materials, such as biopolymers and essential oils, could be a sustainable and eco-friendly alternative for protecting and preserving agricultural products. In this work, we developed films and coatings from pectin and chitosan to protect strawberries (Fragaria x ananassa Duch.) from spoilage and microbial contamination. We developed three coatings containing equal amounts of glycerol and Sicilian lemon essential oil (LEO) nanoemulsion. We identified seventeen chemicals from LEO by GC-MS chromatogram, including d-limonene, α-Pinene, ß-Pinene, and γ-Terpinene. The pectin and chitosan coatings were further characterized using different physicochemical, mechanical, and biological methods. The films demonstrated satisfactory results in strength and elongation at the perforation as fruit packaging. In addition, the coatings did not influence the weight and firmness of the strawberry pulps. We observed that 100 % essential oil was released in 1440 min resulting from the erosion process. Also, the oil preserved the chemical stability of the films. Antioxidant activity (AA), measured by Electron Paramagnetic Resonance (EPR), showed that the coatings loaded with 2 % LEO nanoemulsion (PC + oil) showed that almost 50 % of AA from LEO nanoemulsion was preserved. The chitosan and the pectin-chitosan coatings (PC + oil) inhibited filamentous fungi and yeast contaminations in strawberries for at least 14 days, showing a relationship between the AA and antimicrobial results.
Asunto(s)
Quitosano , Fragaria , Aceites Volátiles , Aceites Volátiles/farmacología , Aceites Volátiles/química , Fragaria/microbiología , Quitosano/química , Pectinas/farmacología , Pectinas/química , Antioxidantes/farmacología , Antioxidantes/química , Conservación de Alimentos/métodosRESUMEN
Engineered human tissues created by three-dimensional cell culture of human cells in a hydrogel are becoming emerging model systems for cancer drug discovery and regenerative medicine. Complex functional engineered tissues can also assist in the regeneration, repair, or replacement of human tissues. However, one of the main hurdles for tissue engineering, three-dimensional cell culture, and regenerative medicine is the capability of delivering nutrients and oxygen to cells through the vasculatures. Several studies have investigated different strategies to create a functional vascular system in engineered tissues and organ-on-a-chips. Engineered vasculatures have been used for the studies of angiogenesis, vasculogenesis, as well as drug and cell transports across the endothelium. Moreover, vascular engineering allows the creation of large functional vascular conduits for regenerative medicine purposes. However, there are still many challenges in the creation of vascularized tissue constructs and their biological applications. This review will summarize the latest efforts to create vasculatures and vascularized tissues for cancer research and regenerative medicine.
RESUMEN
Psoriasis is a disease that causes keratinocytes to proliferate ten times faster than normal, resulting in chronic inflammation and immune cell infiltration in the skin. Aloe vera (A. vera) creams have been used topically for treating psoriasis because they contain several antioxidant species; however, they have several limitations. Natural rubber latex (NRL) has been used as occlusive dressings to promote wound healing by stimulating cell proliferation, neoangiogenesis, and extracellular matrix formation. In this work, we developed a new A. vera-releasing NRL dressing by a solvent casting method to load A. vera into NRL. FTIR and rheological analyzes revealed no covalent interactions between A. vera and NRL in the dressing. We observed that 58.8 % of the loaded A. vera, present on the surface and inside the dressing, was released after 4 days. Biocompatibility and hemocompatibility were validated in vitro using human dermal fibroblasts and sheep blood, respectively. We observed that ~70 % of the free antioxidant properties of A. vera were preserved, and the total phenolic content was 2.31-fold higher than NRL alone. In summary, we combined the antipsoriatic properties of A. vera with the healing activity of NRL to generate a novel occlusive dressing that may be indicated for the management and/or treatment of psoriasis symptoms simply and economically.
Asunto(s)
Aloe , Psoriasis , Humanos , Animales , Ovinos , Goma , Látex , Antioxidantes/farmacología , Psoriasis/tratamiento farmacológico , VendajesRESUMEN
Soft tissue defects are a common clinical challenge mostly caused by trauma, congenital anomalies and oncological surgery. Current soft tissue reconstruction options include synthetic materials (fillers and implants) and autologous adipose tissue transplantation through flap surgery and/or lipotransfer. Both reconstructive options hold important disadvantages to which vascularized adipose tissue engineering (VATE) strategies could offer solutions. In this review, we first summarized pivotal characteristics of functional adipose tissue such as the structure, function, cell types, development and extracellular matrix (ECM). Next, we discussed relevant cell sources and how they are applied in different state-of-the-art VATE techniques. Herein, biomaterial scaffolds and hydrogels, ECMs, spheroids, organoids, cell sheets, three dimensional printing and microfluidics are overviewed. Also, we included extracellular vesicles and emphasized their potential role in VATE. Lastly, current challenges and future perspectives in VATE are pointed out to help to pave the road towards clinical applications.
Asunto(s)
Ingeniería de Tejidos , Andamios del Tejido , Andamios del Tejido/química , Ingeniería de Tejidos/métodos , Tejido Adiposo , Materiales Biocompatibles , HidrogelesRESUMEN
Copper chalcogenide Cu2(Se,Te) compounds are well known as typical p-type thermoelectric materials with a figure of merit (ZT) that can be optimized by the ratio of Se : Te. Here, by using the mechanical alloying and solid-state reaction methods, Te was substituted into Se sites within Cu2Se as the formula Cu2Se1-x Te x (x = 0.1, 0.2, 0.25, and 0.3). The observed changes in structural phase, grain morphologies, and grain size were recorded by XRD and FE-SEM imaging with the appearance of the secondary phase of Cu2Te, with a Te content of x = 0.25. The layered structure morphology was observed more clearly at the high Te content. The electrical conductivity was greatly increased with enriched Te content while the maximum Seebeck coefficient was obtained in the Cu2Se0.75Te0.25 sample. Accordingly, a power factor value of up to 9.84 µW cm-1 K-2 at 773 K was achieved. The appearance of a Cu2Te phase with a Te content of 0.25 created a structural phase transition which results in a ZT value of 1.35 at 773 K in the Cu2Se0.75Te0.25 sample.
RESUMEN
The human brain and central nervous system (CNS) present unique challenges in drug development for neurological diseases. One major obstacle is the blood-brain barrier (BBB), which hampers the effective delivery of therapeutic molecules into the brain while protecting it from blood-born neurotoxic substances and maintaining CNS homeostasis. For BBB research, traditional in vitro models rely upon Petri dishes or Transwell systems. However, these static models lack essential microenvironmental factors such as shear stress and proper cell-cell interactions. To this end, organ-on-a-chip (OoC) technology has emerged as a new in vitro modeling approach to better recapitulate the highly dynamic in vivo human brain microenvironment so-called the neural vascular unit (NVU). Such BBB-on-a-chip models have made substantial progress over the last decade, and concurrently there has been increasing interest in modeling various neurological diseases such as Alzheimer's disease and Parkinson's disease using OoC technology. In addition, with recent advances in other scientific technologies, several new opportunities to improve the BBB-on-a-chip platform via multidisciplinary approaches are available. In this review, an overview of the NVU and OoC technology is provided, recent progress and applications of BBB-on-a-chip for personalized medicine and drug discovery are discussed, and current challenges and future directions are delineated.
Asunto(s)
Enfermedad de Alzheimer , Barrera Hematoencefálica , Transporte Biológico , Encéfalo , Humanos , Dispositivos Laboratorio en un ChipRESUMEN
The main purpose of this paper is the study of the correlations between Image Aesthetic (IA) and Image Naturalness (IN) and the analysis of the influence of IA and IN on Image Quality (IQ) in different contexts. The first contribution is a study about the potential relationships between IA and IN. For that study, two sub-questions are considered. The first one is to validate the idea that IA and IN are not correlated to each other. The second one is about the influence of IA and IN features on Image Naturalness Assessment (INA) and Image Aesthetic Assessment (IAA), respectively. Secondly, it is obvious that IQ is related to IA and IN, but the exact influence of IA and IN on IQ has not been evaluated. Besides that, the context impact on those influences has not been clarified, so the second contribution is to investigate the influence of IA and IN on IQ in different contexts. The results obtained from rigorous experiments prove that although there are moderate and weak correlations between IA and IN, they are still two different components of IQ. It also appears that viewers' IQ perception is affected by some contextual factors, and the influence of IA and IN on IQ depends on the considered context.
RESUMEN
Micropost-based microfluidic devices are widely used for microvascular network (MVN) formation in diverse research fields. However, consistently generating perfusable MVNs of physiological morphology and dimension has proven to be challenging. Here, how initial seeding parameters determine key characteristics of MVN formation is investigated and a robust two-step seeding strategy to generate perfusable physiological MVNs in microfluidic devices is established.
Asunto(s)
Microvasos , Neovascularización Fisiológica , Dispositivos Laboratorio en un Chip , Neovascularización Fisiológica/fisiologíaRESUMEN
We report a five-fold luminance increase of green-light-emitting CdSe@ZnS quantum-dot LEDs (QLEDs) in response to treatment with a 2-ethoxyethanol solution of cesium carbonate (Cs2CO3). The maximum luminous yield of Cs2CO3-treated QLED is as high as 3.41 cd A-1 at 6.4 V. To elucidate device-performance improvement, we model measured currents as the sum of radiative and non-radiative recombination components, which are respectively represented by modified Shockley equations. Variations in model parameters show that a shift in Fermi level, reduction of barrier heights, and passivation of mid-gap defect states are the main results of Cs2CO3 treatment. In spite of a large luminance difference, light-extraction efficiency remains the same at 9% regardless of Cs2CO3 treatment because of the similarity in optical structures.
RESUMEN
This review describes recent research that has advanced our understanding of the role of immune cells in the tumor microenvironment (TME) using advanced 3D in vitro models and engineering approaches. The TME can hinder effective eradication of tumor cells by the immune system, but immunotherapy has been able to reverse this effect in some cases. However, patient-to-patient variability in response suggests that we require deeper understanding of the mechanistic interactions between immune and tumor cells to improve response and develop novel therapeutics. Reconstruction of the TME using engineered 3D models allows high-resolution observation of cell interactions while allowing control of conditions such as hypoxia, matrix stiffness, and flow. Moreover, patient-derived organotypic models are an emerging tool for prediction of drug efficacy. This review highlights the importance of modeling and understanding the immune TME and describes new tools for identifying new biological targets, drug testing, and strategies for personalized medicine.
RESUMEN
Microfluidic technology has been extensively applied to model the functional units of human organs and tissues. Since vasculature is a key component of any functional tissue, a variety of techniques to mimic vasculature in vitro have been developed to address complex physiological and pathological processes in 3D tissues. Herein, we developed a novel, in vitro, microfluidic-based model to probe microvasculature growth into and across implanted porous membranes. Using ePTFE and polycarbonate as examples, we characterize the vascularization potential of these thin porous membranes using this device. This tool will allow for the assessment of porous materials early in their development, prior to their use for encapsulating implants or drugs, while minimizing the need for animal studies. Employing quantitative morphometric analysis and measurements of vascular permeability, we demonstrate our model to be an effective platform for evaluation of angiogenic potential of an implanted membrane biomaterial. Results show that endothelial cells can either migrate as single cells or form continuous sprouts across porous membranes, which is a material structure-dependent behavior. Our model is advantageous over conventional Transwell assays as it is amenable to quantitative assessment of vascular sprouting in 3D, and in contrast to animal models it can be employed more efficiently and with real-time assessment capabilities. This new tool could be applied either to test the suitability of a wide range of biomaterials for implantation or to screen different pro-angiogenic factors for therapeutic applications, and will advance the design of new biomaterials.
Asunto(s)
Células Endoteliales , Neovascularización Patológica , Animales , Materiales Biocompatibles , Humanos , Microvasos , Neovascularización Fisiológica , PorosidadRESUMEN
The main goal of this paper is to study Image Aesthetic Assessment (IAA) indicating images as high or low aesthetic. The main contributions concern three points. Firstly, following the idea that photos in different categories (human, flower, animal, landscape, ) are taken with different photographic rules, image aesthetic should be evaluated in a different way for each image category. Large field images and close-up images are two typical categories of images with opposite photographic rules so we want to investigate the intuition that prior Large field/Close-up Image Classification (LCIC) might improve the performance of IAA. Secondly, when a viewer looks at a photo, some regions receive more attention than other regions. Those regions are defined as Regions Of Interest (ROI) and it might be worthy to identify those regions before IAA. The question "Is it worthy to extract some ROIs before IAA?" is considered by studying Region Of Interest Extraction (ROIE) before investigating IAA based on each feature set (global image features, ROI features and background features). Based on the answers, a new IAA model is proposed. The last point is about a comparison between the efficiency of handcrafted and learned features for the purpose of IAA.
RESUMEN
We report on a substantial increase in luminance and luminous efficiency of green-light emitting devices (LEDs) that use colloidal CdSe@ZnS quantum dots (QDs) as a light-emitting material in response to treatment with 1,2-ethanedithiol (EDT). The maximum luminance increased from 1146 to 8075 cd m-2, and luminous yield from 0.15 to 1.41 cd A-1 as a result of treating an incomplete device with drops of EDT right after spin-coating QDs onto a ZnO-nanoparticle layer. Based on systematic studies on substrate-dependent change in photoluminescence, and current-voltage and luminance-voltage characteristics, we propose that passivation of intra-gap defect states and relative shifts of energy levels relevant to the operation of QD LEDs are two main results of EDT treatment. In particular, we argue that energy-level shift without emission-color change can be attributed to surface-dipole effects.
RESUMEN
Immunohistochemistry and fluorescence in situ hybridization are the two standard methods for human epidermal growth factor receptor 2 (HER2) assessment. However, they have severe limitations to assess quantitatively intratumoral heterogeneity (ITH) when multiple subclones of tumor cells co-exist. We develop here a high-content, quantitative analysis of breast cancer tissues based on microfluidic experimentation and image processing, to characterize both HER2 protein overexpression and HER2 gene amplification at the cellular level. The technique consists of performing sequential steps on the same tissue slide: an immunofluorescence (IF) assay using a microfluidic protocol, an elution step for removing the IF staining agents, a standard FISH staining protocol, followed by automated quantitative cell-by-cell image processing. Moreover, ITH is accurately detected in both cluster and mosaic form using an analysis of spatial association and a mathematical model that allows discriminating true heterogeneity from artifacts due to the use of thin tissue sections. This study paves the way to evaluate ITH with high accuracy and content while requiring standard staining methods.
Asunto(s)
Neoplasias de la Mama/genética , Amplificación de Genes , Regulación Neoplásica de la Expresión Génica , Heterogeneidad Genética , Receptor ErbB-2/genética , Neoplasias de la Mama/metabolismo , Neoplasias de la Mama/patología , Línea Celular Tumoral , Femenino , Técnica del Anticuerpo Fluorescente/métodos , Humanos , Hibridación Fluorescente in Situ/métodos , Técnicas Analíticas Microfluídicas/métodos , Receptor ErbB-2/metabolismo , Reproducibilidad de los ResultadosRESUMEN
We report systematic efficiency variations of green-emitting CdSe@ZnS quantum-dot (QD) LEDs (QLEDs) in response to in situ treatments with 1,2-ethanedithiol (EDT) solutions at various concentrations. The main effect of in situ EDT treatment on a QD layer spin-coated onto a ZnO layer was vacuum-level shift due to dipole moments on the surface of the QD layer and at the interface between QD and ZnO layers. Competing contributions of these dipole moments were responsible for changes in energy level configurations and, accordingly, electron and hole barriers that resulted in discrepancies in electron- and hole-current variations. QLED efficiency was best when treated with an EDT solution of 4 mM, attributable to the largest increase in the hole- to electron current ratio. The maximum luminous yield of the 4 mM EDT-treated QLED was 5.43 cd A-1, which is 10 times higher than that of an untreated device. Furthermore, the luminous yield of this treated device remained as high as 2.56 cd A-1 at a luminance of 500 cd m-2.