31 results on '"Carolina Lucchesi"'
Search Results
2. A Novel Microphysiological Colon Platform to Decipher Mechanisms Driving Human Intestinal PermeabilitySummary
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Athanasia Apostolou, Rohit A. Panchakshari, Antara Banerjee, Dimitris V. Manatakis, Maria D. Paraskevopoulou, Raymond Luc, Galeb Abu-Ali, Alexandra Dimitriou, Carolina Lucchesi, Gauri Kulkarni, Tengku Ibrahim Maulana, Magdalena Kasendra, Jordan S. Kerns, Bertram Bleck, Lorna Ewart, Elias S. Manolakos, Geraldine A. Hamilton, Cosmas Giallourakis, and Katia Karalis more...
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Organoids ,Organ-on-Chip ,Leaky Gut ,Interleukin 22 ,Diseases of the digestive system. Gastroenterology ,RC799-869 - Abstract
Background & Aims: The limited availability of organoid systems that mimic the molecular signatures and architecture of human intestinal epithelium has been an impediment to allowing them to be harnessed for the development of therapeutics as well as physiological insights. We developed a microphysiological Organ-on-Chip (Emulate, Inc, Boston, MA) platform designed to mimic properties of human intestinal epithelium leading to insights into barrier integrity. Methods: We combined the human biopsy-derived leucine-rich repeat-containing G-protein–coupled receptor 5–positive organoids and Organ-on-Chip technologies to establish a micro-engineered human Colon Intestine-Chip (Emulate, Inc, Boston, MA). We characterized the proximity of the model to human tissue and organoids maintained in suspension by RNA sequencing analysis, and their differentiation to intestinal epithelial cells on the Colon Intestine-Chip under variable conditions. Furthermore, organoids from different donors were evaluated to understand variability in the system. Our system was applied to understanding the epithelial barrier and characterizing mechanisms driving the cytokine-induced barrier disruption. Results: Our data highlight the importance of the endothelium and the in vivo tissue-relevant dynamic microenvironment in the Colon Intestine-Chip in the establishment of a tight monolayer of differentiated, polarized, organoid-derived intestinal epithelial cells. We confirmed the effect of interferon-γ on the colonic barrier and identified reorganization of apical junctional complexes, and induction of apoptosis in the intestinal epithelial cells as mediating mechanisms. We show that in the human Colon Intestine-Chip exposure to interleukin 22 induces disruption of the barrier, unlike its described protective role in experimental colitis in mice. Conclusions: We developed a human Colon Intestine-Chip platform and showed its value in the characterization of the mechanism of action of interleukin 22 in the human epithelial barrier. This system can be used to elucidate, in a time- and challenge-dependent manner, the mechanism driving the development of leaky gut in human beings and to identify associated biomarkers. more...
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- 2021
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Catalog
3. Author Correction: Performance assessment and economic analysis of a human Liver-Chip for predictive toxicology
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Lorna Ewart, Athanasia Apostolou, Skyler A. Briggs, Christopher V. Carman, Jake T. Chaff, Anthony R. Heng, Sushma Jadalannagari, Jeshina Janardhanan, Kyung-Jin Jang, Sannidhi R. Joshipura, Mahika M. Kadam, Marianne Kanellias, Ville J. Kujala, Gauri Kulkarni, Christopher Y. Le, Carolina Lucchesi, Dimitris V. Manatakis, Kairav K. Maniar, Meaghan E. Quinn, Joseph S. Ravan, Ann Catherine Rizos, John F. K. Sauld, Josiah D. Sliz, William Tien-Street, Dennis Ramos Trinidad, James Velez, Max Wendell, Onyi Irrechukwu, Prathap Kumar Mahalingaiah, Donald E. Ingber, Jack W. Scannell, and Daniel Levner more...
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Medicine - Published
- 2023
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4. Duodenum Intestine-Chip for preclinical drug assessment in a human relevant model
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Magdalena Kasendra, Raymond Luc, Jianyi Yin, Dimitris V Manatakis, Gauri Kulkarni, Carolina Lucchesi, Josiah Sliz, Athanasia Apostolou, Laxmi Sunuwar, Jenifer Obrigewitch, Kyung-Jin Jang, Geraldine A Hamilton, Mark Donowitz, and Katia Karalis more...
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Organs-on-Chips ,organoids ,duodenum ,CYP3A4 ,drug transport ,drug metabolism ,Medicine ,Science ,Biology (General) ,QH301-705.5 - Abstract
Induction of intestinal drug metabolizing enzymes can complicate the development of new drugs, owing to the potential to cause drug-drug interactions (DDIs) leading to changes in pharmacokinetics, safety and efficacy. The development of a human-relevant model of the adult intestine that accurately predicts CYP450 induction could help address this challenge as species differences preclude extrapolation from animals. Here, we combined organoids and Organs-on-Chips technology to create a human Duodenum Intestine-Chip that emulates intestinal tissue architecture and functions, that are relevant for the study of drug transport, metabolism, and DDI. Duodenum Intestine-Chip demonstrates the polarized cell architecture, intestinal barrier function, presence of specialized cell subpopulations, and in vivo relevant expression, localization, and function of major intestinal drug transporters. Notably, in comparison to Caco-2, it displays improved CYP3A4 expression and induction capability. This model could enable improved in vitro to in vivo extrapolation for better predictions of human pharmacokinetics and risk of DDIs. more...
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- 2020
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5. Lithograph-moulded poly-L-co-D,L lactide porous membranes for osteoblastic culture
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Andre D. Messias, Carolina Lucchesi, Débora C. Coraça-Huber, Aristides Pavani Filho, and Eliana A. R. Duek
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bone tissue engineering ,cell adhesion ,lithography ,osteoblastic cells ,poly-L-co-D,L lactide ,porosity ,viability ,Materials of engineering and construction. Mechanics of materials ,TA401-492 - Abstract
Pore size, shape, wall morphology, porosity, and interconnectivity are important characteristics of the scaffolds. Lithography is a manufacturing technique that allows the production of tridimensional scaffolds with a controllable and reproducible inner architecture. The aim of this study was to use lithography to create a poly-L-co-D,L lactide (PLDLA) scaffold with symmetrical pore size and distribution, and to evaluate its biocompatibility with osteoblasts in vitro. Lithographic moulds were used to produce porous PLDLA membranes by a casting procedure. Osteoblasts were removed from calvarial bones and seeded onto porous and smooth PLDLA membranes after which cell viability and adhesion assays, cytochemical analysis and scanning electron microscopy were used to characterize the cells. Cell viability and adhesion assays, cytochemical analysis, and scanning electron microscopy were carried out. Cell viability was similar on porous and smooth PLDLA membranes but higher than on a polystyrene substrate (positive control). Although osteoblasts adhered to the surface of all the materials tested, cell adhesion to lithographed PLDLA was greater than to smooth PLDLA membranes. In conclusion, osteoblasts interacted well with PLDLA membranes, as shown by the viability and adhesion assays and by the enhanced collagen production. more...
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- 2013
6. Benefits of oxygen and nitrogen plasma treatment in Vero cell affinity to poly(lactide-co-glycolide acid)
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Andrea Rodrigues Esposito, Camila Mika Kamikawa, Carolina Lucchesi, Betina Mara Pereira Ferreira, and Eliana Aparecida de Rezende Duek
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biomaterials ,plasma treatment ,PLGA ,cell culture ,Materials of engineering and construction. Mechanics of materials ,TA401-492 - Abstract
Cell adhesion on materials surface is critical because this phenomenon occurs before other events, as cell spreading, cell migration and cell differentiation. it is commonly accepted that the adhesion of cells on solid substrate is influenced by several substratum surface properties, such as wettability, surface charge, roughness and topography. plasma technique is a convenient method for modifying surface properties of materials without affecting physical properties. in this study, poly(lactide-co-glycolide), plga, membranes were modified by oxygen and nitrogen plasma to improve polymer hydrophilicity and verify their effect on vero cells culture. the plga membranes, which were characterized by sem and contact angle, showed increased surface rugosity and narrower contact angles. cell adhesion, cytotoxicity assay, sem and cytochemistry analysis showed that plasma treatment was beneficial to cell growth by improving cell-polymer interaction. Cell adhesion on materials surface is critical because this phenomenon occurs before other events, as cell spreading, cell migration and cell differentiation. It is commonly accepted that the adhesion of cells on solid substrate is influenced by several substratum surface properties, such as wettability, surface charge, roughness and topography. Plasma technique is a convenient method for modifying surface properties of materials without affecting physical properties. In this study, poly(lactide-co-glycolide), PLGA, membranes were modified by oxygen and nitrogen plasma to improve polymer hydrophilicity and verify their effect on Vero cells culture. The PLGA membranes, which were characterized by SEM and contact angle, showed increased surface rugosity and narrower contact angles. Cell adhesion, cytotoxicity assay, SEM and cytochemistry analysis showed that plasma treatment was beneficial to cell growth by improving cell-polymer interaction. more...
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- 2013
7. Citocompatibilidade de blendas de poli(p-dioxanona)/ poli(hidroxi butirato) (PPD/PHB) para aplicações em engenharia de tecido cartilaginoso Cytocompatibility of poly(p-dioxanone)/ poly(hydroxybutic) (PPD/PHB) blends to cartilage tissue engineering
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Andrea R. Esposito, Eliana A. R. Duek, Carolina Lucchesi, Luciana Prazeres, and Ana P. T. Pezzin
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Blenda ,poli(p-dioxanona) ,poli(hidroxi butirato) ,fibrocondrócitos ,Blend ,poly(p-dioxanone) ,poly(hydroxybutic) ,fibrochondrocytes ,Chemical technology ,TP1-1185 - Abstract
Buscando estratégias que repercutam na melhoria da interação entre materiais poliméricos biorreabsorvíveis e o crescimento celular, o presente estudo in vitro teve como objetivo estudar a influência de blendas de PPD/PHB na adesão celular e crescimento de fibrocondrócitos obtidos a partir de cultura primária. As blendas de PPD/PHB foram preparadas pelo método de evaporação de solvente nas composições 100/0, 60/40 e 50/50 e caracterizadas por microscopia eletrônica de varredura (MEV). Observações ultra-estruturais mostraram alterações na morfologia celular, sugerindo que os fibrocondrócitos podem responder a alterações no substrato alterando seu perfil fenotípico. As análises com MTT demonstraram que as blendas não apresentaram citotoxicidade e permitiram a adesão e proliferação dos fibrocondrócitos sobre os substratos em todas as suas composições. O ensaio colorimétrico com Sirius Red evidenciou a capacidade de manutenção da síntese de matriz extracelular colágena sobre as amostras, concluindo-se que as blendas de PPD/PHB podem ser indicadas para o cultivo celular.In order of seek strategies to improve the interaction between bioreabsorbable polymer materials and cellular growth, this work aimed at evaluating in vitro the influence of PPD/PHB blends on cell adhesion and fibrochondrocytes growth. Fibrochondrocytes cells were obtained by primary extraction from enzymatic digestion methods. The PPD/PHB blends were prepared by casting with 100/0, 60/40 and 50/50 compositions, and were characterized by scanning electron microscopy (SEM). After 6, 48, 120 and 168 hours in culture, ultrastructural observations showed changes in cell morphology, suggesting that the fibrochondrocytes can respond to substrate modifications, changing their phenotypic profile. The MTT analyses showed that the blends did not present cytotoxicity and allowed fibrochondrocytes adhesion and proliferation on the membranes in all compositions. The colorimetric Sirius Red test revealed the capability of extracellular matrix synthesis on the blends, from which one can conclude that the PPD/PHB blends are not cytotoxic and can be indicated for cell culture. more...
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- 2010
8. Cell Adhesion to Plasma-Coated PVC
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Elidiane C. Rangel, Eduardo S. de Souza, Francine S. de Moraes, Eliana A. R. Duek, Carolina Lucchesi, Wido H. Schreiner, Steven F. Durrant, and Nilson C. Cruz
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Technology ,Medicine ,Science - Abstract
To produce environments suitable for cell culture, thin polymer films were deposited onto commercial PVC plates from radiofrequency acetylene-argon plasmas. The proportion of argon in the plasmas, PAr, was varied from 5.3 to 65.8%. The adhesion and growth of Vero cells on the coated surfaces were examined for different incubation times. Cytotoxicity tests were performed using spectroscopic methods. Carbon, O, and N were detected in all the samples using XPS. Roughness remained almost unchanged in the samples prepared with 5.3 and 28.9% but tended to increase for the films deposited with PAr between 28.9 and 55.3%. Surface free energy increased with increasing PAr, except for the sample prepared at 28.9% of Ar, which presented the least reactive surface. Cells proliferated on all the samples, including the bare PVC. Independently of the deposition condition there was no evidence of cytotoxicity, indicating the viability of such coatings for designing biocompatible devices. more...
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- 2014
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9. A Novel Microphysiological Colon Platform to Decipher Mechanisms Driving Human Intestinal Permeability
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Rohit A. Panchakshari, Athanasia Apostolou, Cosmas Giallourakis, Galeb Abu-Ali, Antara Banerjee, Tengku Ibrahim Maulana, Raymond Luc, Gauri Kulkarni, Carolina Lucchesi, Jordan Kerns, Katia Karalis, Alexandra Dimitriou, Maria D. Paraskevopoulou, Magdalena Kasendra, Elias S. Manolakos, Lorna Ewart, Geraldine A. Hamilton, Bertram Bleck, and Dimitris V. Manatakis more...
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EdU, 5-ethynyl-2’-deoxyuridine ,Cell Culture Techniques ,RC799-869 ,Wnt, wingless-related integration site ,IEC, intestinal epithelial cell ,Interleukin 22 ,ZO-1, zonula occludens-1 ,IL, interleukin 6 ,Organ-on-Chip ,Leaky Gut ,Lab-On-A-Chip Devices ,Intestinal Mucosa ,Receptor ,Original Research ,DKK1, Dickkopf-related protein 1 ,IBD, inflammatory bowel disease ,cHIMEC, colonic human intestinal microvascular endothelial cell ,Gastroenterology ,Diseases of the digestive system. Gastroenterology ,Intestinal epithelium ,NHS, N-hydroxysuccinimide ,Cell biology ,ECM, extracellular matrix ,Organoids ,STAT, signal transducer and activator of transcription ,TNFα, tumor necrosis factor α ,medicine.anatomical_structure ,TJ, tight junction ,Cellular Microenvironment ,PDMS, polydimethylsiloxane ,qPCR, quantitative polymerase chain reaction ,Endothelium ,Colon ,PBS, phosphate-buffered saline ,Biology ,Permeability ,In vivo ,medicine ,Organoid ,Humans ,DGE, differential gene expression ,IFNγ, interferon-γ ,GO, gene ontology ,Intestinal permeability ,Hepatology ,Mechanism (biology) ,Gene Expression Profiling ,Interleukins ,RNA-seq, RNA sequencing ,Computational Biology ,medicine.disease ,IL22BP, interleukin 22 binding protein ,Papp, apparent permeability ,Gene Expression Regulation ,Transcriptome ,Biomarkers - Abstract
Background & Aims The limited availability of organoid systems that mimic the molecular signatures and architecture of human intestinal epithelium has been an impediment to allowing them to be harnessed for the development of therapeutics as well as physiological insights. We developed a microphysiological Organ-on-Chip (Emulate, Inc, Boston, MA) platform designed to mimic properties of human intestinal epithelium leading to insights into barrier integrity. Methods We combined the human biopsy-derived leucine-rich repeat-containing G-protein–coupled receptor 5–positive organoids and Organ-on-Chip technologies to establish a micro-engineered human Colon Intestine-Chip (Emulate, Inc, Boston, MA). We characterized the proximity of the model to human tissue and organoids maintained in suspension by RNA sequencing analysis, and their differentiation to intestinal epithelial cells on the Colon Intestine-Chip under variable conditions. Furthermore, organoids from different donors were evaluated to understand variability in the system. Our system was applied to understanding the epithelial barrier and characterizing mechanisms driving the cytokine-induced barrier disruption. Results Our data highlight the importance of the endothelium and the in vivo tissue-relevant dynamic microenvironment in the Colon Intestine-Chip in the establishment of a tight monolayer of differentiated, polarized, organoid-derived intestinal epithelial cells. We confirmed the effect of interferon-γ on the colonic barrier and identified reorganization of apical junctional complexes, and induction of apoptosis in the intestinal epithelial cells as mediating mechanisms. We show that in the human Colon Intestine-Chip exposure to interleukin 22 induces disruption of the barrier, unlike its described protective role in experimental colitis in mice. Conclusions We developed a human Colon Intestine-Chip platform and showed its value in the characterization of the mechanism of action of interleukin 22 in the human epithelial barrier. This system can be used to elucidate, in a time- and challenge-dependent manner, the mechanism driving the development of leaky gut in human beings and to identify associated biomarkers., Graphical abstract more...
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- 2021
10. Combining Human Organoids and Organ-on-a-Chip Technology to Model Intestinal Region-Specific Functionality
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Magdalena Kasendra, Carolina Lucchesi, Lorna Ewart, Athanasia Apostolou, and Gauri Kulkarni
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Organoids ,Technology ,General Immunology and Microbiology ,General Chemical Engineering ,General Neuroscience ,Lab-On-A-Chip Devices ,Endothelial Cells ,Humans ,Intestinal Mucosa ,General Biochemistry, Genetics and Molecular Biology - Abstract
The intestinal mucosa is a complex physical and biochemical barrier that fulfills a myriad of important functions. It enables the transport, absorption, and metabolism of nutrients and xenobiotics while facilitating a symbiotic relationship with microbiota and restricting the invasion of microorganisms. Functional interaction between various cell types and their physical and biochemical environment is vital to establish and maintain intestinal tissue homeostasis. Modeling these complex interactions and integrated intestinal physiology in vitro is a formidable goal with the potential to transform the way new therapeutic targets and drug candidates are discovered and developed. Organoids and Organ-on-a-Chip technologies have recently been combined to generate human-relevant intestine chips suitable for studying the functional aspects of intestinal physiology and pathophysiology in vitro. Organoids derived from the biopsies of the small (duodenum) and large intestine are seeded into the top compartment of an organ chip and then successfully expand as monolayers while preserving the distinct cellular, molecular, and functional features of each intestinal region. Human intestine tissue-specific microvascular endothelial cells are incorporated in the bottom compartment of the organ chip to recreate the epithelial-endothelial interface. This novel platform facilitates luminal exposure to nutrients, drugs, and microorganisms, enabling studies of intestinal transport, permeability, and host-microbe interactions. Here, a detailed protocol is provided for the establishment of intestine chips representing the human duodenum (duodenum chip) and colon (colon chip), and their subsequent culture under continuous flow and peristalsis-like deformations. We demonstrate methods for assessing drug metabolism and CYP3A4 induction in duodenum chip using prototypical inducers and substrates. Lastly, we provide a step-by-step procedure for the in vitro modeling of interferon gamma (IFNγ)-mediated barrier disruption (leaky gut syndrome) in a colon chip, including methods for evaluating the alteration of paracellular permeability, changes in cytokine secretion, and transcriptomic profiling of the cells within the chip. more...
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- 2022
11. Qualifying a human Liver-Chip for predictive toxicology: Performance assessment and economic implications
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Lorna Ewart, Athanasia Apostolou, Skyler A. Briggs, Christopher V. Carman, Jake T. Chaff, Anthony R. Heng, Sushma Jadalannagari, Jeshina Janardhanan, Kyung-Jin Jang, Sannidhi R. Joshipura, Mahika M. Kadam, Marianne Kanellias, Ville J. Kujala, Gauri Kulkarni, Christopher Y. Le, Carolina Lucchesi, Dimitris V. Manatakis, Kairav K. Maniar, Meaghan E. Quinn, Joseph S. Ravan, Ann Catherine Rizos, John F.K. Sauld, Josiah D. Sliz, William Tien-Street, Dennis Ramos Trinidad, James Velez, Max Wendell, Onyi Irrechukwu, Prathap Kumar Mahalingaiah, Donald E. Ingber, Jack W. Scannell, and Daniel Levner more...
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Human organ-on-a-chip (Organ-Chip) technology has the potential to disrupt preclinical drug discovery and improve success in drug development pipelines as it can recapitulate organ-level pathophysiology and clinical responses. The Innovation and Quality (IQ) consortium formed by multiple pharmaceutical and biotechnology companies to confront this challenge has published guidelines that define criteria for qualifying preclinical models, however, systematic and quantitative evaluation of the predictive value of Organ-Chips has not yet been reported. Here, 870 Liver-Chips were analyzed to determine their ability to predict drug-induced liver injury (DILI) caused by small molecules identified as benchmarks by the IQ consortium. The Liver-Chip met the qualification guidelines across a blinded set of 27 known hepatotoxic and non-toxic drugs with a sensitivity of 87% and a specificity of 100%. A computational economic value analysis suggests that with this performance the Liver-Chip could generate $3 billion annually for the pharmaceutical industry due to increased R&D productivity. more...
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- 2021
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12. Optimization of Human Lung-Chip cultures to investigate effects of biomechanics on primary airway epithelial cell biology
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Geraldine A. Hamilton, Sander van Riet, Annemarie van Schadewijk, Janna Nawroth, Carolina Lucchesi, Doris Roth, Pieter S. Hiemstra, and Anne M. van der Does
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Leukocyte migration ,medicine.anatomical_structure ,Cell ,Biomechanics ,medicine ,Respiratory epithelium ,Biology ,Airway ,Epithelium ,Bronchial Epithelial Cell ,Human lung ,Cell biology - Abstract
With advances brought on by Organ-on-Chip technology, a new level of complexity was introduced into cell systems that allows research into human cellular cross-talk combined with mechanical cues. This added complexity is especially relevant in the lungs where biomechanics play a prominent role. Here we leveraged the commercial (EmulateTM) Lung-Chip and optimized its use for primary bronchial epithelial cell (PBEC) culturing to investigate the impact of airflow and mild stretch on epithelial biology. For this, PBEC culturing had to be optimized on a flexible PDMS, 7 µm pore-size membrane that allows application of stretch and leukocyte migration. To obtain a fully differentiated airway epithelium, cell media selection, coating strategy, and prevention of PBEC migration to the bottom channel was successfully achieved. Inter-chip variability (baseline IL-8 levels (ELISA)) was low (coefficient of variation more...
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- 2020
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13. A Micro-engineered Human Colon Intestine-Chip Platform to Study Leaky Barrier
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Geraldine A. Hamilton, Bertram Bleck, Athanasia Apostolou, Carolina Lucchesi, Katia Karalis, Galeb Abu-Ali, Antara Banerjee, Alexandra Dimitriou, Elias S. Manolakos, Tengku Ibrahim Maulana, Gauri Kulkarni, Cosmas Giallourakis, Maria D. Paraskevopoulou, Raymond Luc, Dimitris V. Manatakis, and Rohit A. Panchakshari more...
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Endothelium ,medicine.medical_treatment ,Biology ,Intestinal epithelium ,Cell biology ,Interleukin 22 ,Immune system ,Cytokine ,medicine.anatomical_structure ,Downregulation and upregulation ,Organoid ,medicine ,Interferon gamma ,medicine.drug - Abstract
The intestinal epithelial barrier supports the symbiotic relationship between the microbiota colonizing the intestinal epithelium and the host immune system to maintain homeostasis. Leaky barrier is increasingly recognized as part of the pathogenesis of a number of chronic conditions in addition to inflammatory and infectious diseases. As our understanding on the regulation of the barrier remains limited, effective therapeutic targeting for the compromised barrier is still an unmet need. Here we combined advancements on the organoids and Organ-on-Chip technologies to establish a micro-engineered Colon Intestine-Chip for studying development and regulation of the human intestinal barrier. Our data demonstrate the significance of the endothelium in co-culture with the epithelial cells within a tissue-relevant microenvironment for the establishment of a tight epithelial barrier of polarized cells. Pathway analysis of the RNA sequencing (RNA-Seq), revealed significant upregulation of mechanisms relevant to the maturation of the intestinal epithelium in organoid-derived epithelial cells in co-culture with endothelium as compared to organoids maintained in suspension. We provide evidence that the Colon Intestine-Chip platform responds to interferon gamma (IFNγ), a prototype cytokine utilized to model inflammation-induced barrier disruption, by induction of apoptosis and reorganization of the apical junctional complexes as shown with other systems. We also describe the mechanism of action of interleukin 22 (IL-22) on mature, organoid-derived intestinal epithelial cells that is consistent with barrier disruption. Overall we propose the Colon Intestine-Chip as a promising human organoid-derived platform to decipher mechanisms driving the development of leaky gut in patients and enable their translation for this unmet medical need. more...
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- 2020
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14. 'A novel Organ-Chip system emulates three-dimensional architecture of the human epithelia and allows fine control of mechanical forces acting on it.'
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K. Karalis, Achille Gravanis, Lian Leng, Riccardo Barrile, Geraldine A. Hamilton, Christopher David Hinojosa, Carolina Lucchesi, Antonio Varone, Norman Wen, Josiah Sliz, and Justin Nguyen
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Microchannel ,Materials science ,Three dimensional architecture ,Interface (computing) ,Fluidics ,Chip ,Microfabrication ,Biomedical engineering ,Communication channel ,Micropatterning - Abstract
Successful translation ofin vivoexperimental data to human patients is an unmet need and a bottleneck in the development of effective therapeutics. micro technology aims to address this need with significant advancements reported recently that enable modeling of organ level function. These microengineered chips enable researcher to recreate critical elements such asin vivorelevant tissue-tissue interface, air-liquid interface, and mechanical forces, such as mechanical stretch and fluidic shear stress, are crucial in emulating tissue level functions. Here, we present the development of a new, comprehensive 3D cell-culture system, where we combined our proprietary Organ-Chip technology with recent advantages in three-dimensional organotypic culture. Leveraging microfabrication techniques, we engineered a flexible chip that consists of a channel containing an organotypic epithelium surrounded by two vacuum channels that can be actuated to stretch the hydrogel throughout its thickness. Furthermore, the ceiling of this channel is a removable lid with a built-in microchannel that can be perfused with liquid or air and removed as needed for direct access to the tissue. The floor of this channel is a porous flexible membrane in contact with a microfluidic channel that provides diffusive mass transport to and from the channel. This additional microfluidic channel can be coated with endothelial cells to emulate a blood vessel and capture endothelial interactions. Our results show that the Open-Top Chip design successfully addresses common challenges associated with the Organs-on-Chips technology, including the capability to incorporate a tissue-specific extracellular matrix gel seeded with primary stromal cells, to reproduce the architectural complexity of tissues by micropatterning the gel, that can be extracted for H&E staining. We provide proof-of-concept data on the feasibility of the system using skin and alveolar epithelial primary cells and by simulating alveolar inflammation. more...
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- 2020
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15. A Microengineered Airway Lung Chip Models Key Features of Viral-induced Exacerbation of Asthma
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Abhishek Shukla, Geraldine A. Hamilton, Hyun-Hee Lee, Carolina Lucchesi, Katia Karalis, Michael Salmon, Shroff Tanvi, Justin Ngyuen, Janna Nawroth, Antonio Varone, Remi Villenave, Stephen E. Alves, and Deion Cheng more...
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Pulmonary and Respiratory Medicine ,Exacerbation ,Rhinovirus ,Clinical Biochemistry ,Bioengineering ,medicine.disease_cause ,Transepithelial Migration ,Models, Biological ,Receptors, Interleukin-8B ,Immune system ,Cytopathogenic Effect, Viral ,Cell Movement ,Lab-On-A-Chip Devices ,medicine ,Humans ,Molecular Biology ,Lung ,Cells, Cultured ,Asthma ,business.industry ,Cell Biology ,respiratory system ,medicine.disease ,respiratory tract diseases ,medicine.anatomical_structure ,Neutrophil Infiltration ,Immunology ,Disease Progression ,Respiratory epithelium ,Microtechnology ,Airway ,business - Abstract
Viral-induced exacerbation of asthma remains a major cause of hospitalization and mortality. New human-relevant models of the airways are urgently needed to understand how respiratory infections may trigger asthma attacks and to advance treatment development. Here, we describe a new human-relevant model of rhinovirus-induced asthma exacerbation that recapitulates viral infection of asthmatic airway epithelium and neutrophil transepithelial migration, and enables evaluation of immunomodulatory therapy. Specifically, a microengineered model of fully differentiated human mucociliary airway epithelium was stimulated with IL-13 to induce a T-helper cell type 2 asthmatic phenotype and infected with live human rhinovirus 16 (HRV16) to reproduce key features of viral-induced asthma exacerbation. We observed that the infection with HRV16 replicated key hallmarks of the cytopathology and inflammatory responses observed in human airways. Generation of a T-helper cell type 2 microenvironment through exogenous IL-13 stimulation induced features of asthmatic airways, including goblet cell hyperplasia, reduction of cilia beating frequency, and endothelial activation, but did not alter rhinovirus infectivity or replication. High-resolution kinetic analysis of secreted inflammatory markers revealed that IL-13 treatment altered IL-6, IFN-λ1, and CXCL10 secretion in response to HRV16. Neutrophil transepithelial migration was greatest when viral infection was combined with IL-13 treatment, whereas treatment with MK-7123, a CXCR2 antagonist, reduced neutrophil diapedesis in all conditions. In conclusion, our microengineered Airway Lung-Chip provides a novel human-relevant platform for exploring the complex mechanisms underlying viral-induced asthma exacerbation. Our data suggest that IL-13 may impair the hosts' ability to mount an appropriate and coordinated immune response to rhinovirus infection. We also show that the Airway Lung-Chip can be used to assess the efficacy of modulators of the immune response. more...
- Published
- 2020
16. Modeling Inflammatory Immune Cell Recruitment and Response on Human Colon Intestine-Chip
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Christopher Carman, Marianne Kanellias, Dennis Ramos, Kairav Maniar, John Saud, Carolina Lucchesi, Gauri Kulkarni, Athanasia Apostulou, Ville J Kujala, and Lorna Ewart
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Immunology ,Immunology and Allergy - Abstract
Objective Immune cell recruitment into tissues is an essential step in inflammatory responses. This occurs in a highly tissue- and stimulus-specific manner, which presents a significant challenge to modeling disease and testing therapeutics ex vivo. We previously developed an advanced primary human vascularized Colon Intestine-Chip model and showed that it recapitulates physiologic cell composition, morphology and barrier function. The goal of this study was to test the ability of this system to model inflammatory bowel disease (IBD)-like immune cell reactions ex vivo. Methods We perfused primary human peripheral blood mononuclear cells (PBMC) across the vascular channel in untreated ‘resting’ or TNFα/chemokine-treated ‘inflamed’ Colon Intestine-Chips. We analyzed total cell recruitment, inflammatory cytokine secretion and barrier function in the following 24–72 hours. Results We show that the perfused PBMC efficiently adhered and transmigrated to the epithelial channel in an inflammation-specific manner. This was followed by an accumulation of proinflammatory cytokines characteristic of IBD (e.g., INFγ, IL1β, IL18), as well as loss of barrier function, the hallmark feature of IBD. We further showed that 1) the recruited cells were strongly enriched in the ‘gut trophic’ α4β7+/CCR9+ PBMC subsets and 2) this recruitment could be blocked with the IBD therapeutic Entyvio, which targets the α4β7-MAdCAM-1 interaction. Conclusion Our findings indicate that our Colon Intestine-Chip can model inflammatory immune cell recruitment and in situ immune reactions that reflect key clinical correlates of IBD. This model may prove effective for development of new anti-inflammatory therapeutics for human intestinal diseases. Supported by Emulate Bio more...
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- 2022
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17. A Micro-Engineered Airway Lung-Chip Models Key Features of Viral-Induced Exacerbation of Asthma
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Janna Nawroth, Shroff Tanvi, Remi Villenave, H-H. Lee, Carolina Lucchesi, Stephen E. Alves, Geraldine A. Hamilton, J. Ngyuen, K. Karalis, Abhishek Shukla, D. Cheng, and Michael Salmon
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Lung ,Exacerbation ,business.industry ,medicine.disease ,medicine.disease_cause ,Endothelial activation ,medicine.anatomical_structure ,Immune system ,Immunology ,Medicine ,Respiratory epithelium ,CXCL10 ,Rhinovirus ,business ,Asthma - Abstract
RationaleViral-induced exacerbation of asthma remain a major cause of hospitalization and mortality. New human relevant models of the airways are urgently needed to understand how respiratory infections may trigger asthma attacks, and to advance treatment development.ObjectivesTo develop a new human relevant model of rhinovirus-induced asthma exacerbation that recapitulates viral infection of asthmatic airway epithelium, neutrophil transepithelial migration, and enables evaluation of immunomodulatory therapy.MethodsA micro-engineered model of fully differentiated human mucociliary airway epithelium was stimulated with IL-13 to induce a Th2-type asthmatic phenotype and infected with live human rhinovirus 16 (HRV16) to reproduce key features of viral-induced asthma exacerbation.Measurements and Main ResultsInfection with HRV16 replicated key hallmarks of the cytopathology and inflammatory responses observed in human airways. Generation of a Th2 microenvironment through exogenous IL-13 stimulation induced features of asthmatics airways, including goblet cell hyperplasia, reduction of cilia beating frequency, and endothelial activation, but did not alter rhinovirus infectivity or replication. High resolution kinetic analysis of secreted inflammatory markers revealed that IL-13 treatment altered the IL-6, IFN-λ1, and CXCL10 secretion in response to HRV16. Neutrophil transepithelial migration was greatest when viral infection was combined with IL-13 treatment, while treatment with MK-7123, a CXCR2 antagonist, reduced neutrophil diapedesis in all conditions.ConclusionsThis micro-engineered Airway Lung-Chip provides a novel human-relevant platform for exploring the complex mechanisms underlying viral-induced asthma exacerbation. Our data suggest that IL-13 may impair the hosts’ ability to mount an appropriate and coordinated immune response to rhinovirus infection. We also show that the Airway Lung-Chip can be used to assess the efficacy of modulators of the immune response.NoteEmulate®, Human Emulation System®, S-1™, ER-1™, and ER-2™ are trademarks of Emulate, Inc., and any other trademarks used herein remain with their respective holders. The technology disclosed in this document may be covered by one or more patents or patent applications, and no license to these is granted herein. You are solely responsible for determining whether you have all intellectual property rights that are necessary for your intended use of any of the disclosed materials, and whether you are required to obtain any additional intellectual property rights from a third party. Further information is available by contacting the authors.At a Glance CommentaryScientific Knowledge on the SubjectNew therapies for asthma exacerbations remain a significant unmet medical need. Development of human relevant preclinical models are needed to further elucidate the complex mechanisms underlying asthma exacerbation and investigate new therapeutic strategies.What This Study Adds to the FieldUsing a human Airway Lung-Chip model, we show here for the first time a live human rhinovirus (HRV) infection of the asthmatic epithelium that recapitulates complex features of viral-induced asthma exacerbation. The dynamic microenvironment of the chip enables the real-time study of virus infection, epithelial response, and immune cell recruitment under healthy and asthmatic conditions. The model reproduces key endpoints that have been observed in asthmatics and individuals infected with rhinovirus including the ciliated cell sloughing, altered cilia beating frequency, goblet cell hyperplasia, increased expression of adhesion molecules in microvascular endothelial cells, and inflammatory mediator release. High-resolution temporal analysis of secreted inflammatory markers enabled by dynamic sampling revealed alteration of IL-6, IFN-λ1 and CXCL10 secretory phases after rhinovirus infection in an IL-13 high environment. Leveraging high-content imaging and analysis of circulating inflammatory cells, we demonstrated the efficacy of a CXCR2 antagonist to reduce adhesion, motility, and transmigration of perfused human neutrophils. Thus, this micro-engineered chip may offer a powerful addition to preclinical models for understanding mechanisms underlying asthma exacerbation pathology and developing new therapeutic strategies. more...
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- 2020
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18. Duodenum Intestine-Chip for preclinical drug assessment in a human relevant model
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Athanasia Apostolou, Geraldine A. Hamilton, Laxmi Sunuwar, Carolina Lucchesi, Katia Karalis, Josiah Sliz, Dimitris V. Manatakis, Mark Donowitz, Jenifer Obrigewitch, Gauri Kulkarni, Raymond Luc, Kyung-Jin Jang, Jianyi Yin, and Magdalena Kasendra more...
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0301 basic medicine ,CYP3A4 ,Drug Evaluation, Preclinical ,duodenum ,02 engineering and technology ,Pharmacology ,Organs-on-Chips ,Cytochrome P-450 CYP3A ,Drug Interactions ,Biology (General) ,organoids ,Barrier function ,media_common ,Microvilli ,General Neuroscience ,General Medicine ,021001 nanoscience & nanotechnology ,medicine.anatomical_structure ,Medicine ,0210 nano-technology ,Research Article ,Human ,Drug ,drug transport ,ATP Binding Cassette Transporter, Subfamily B ,QH301-705.5 ,Science ,media_common.quotation_subject ,Biology ,Permeability ,General Biochemistry, Genetics and Molecular Biology ,03 medical and health sciences ,Organ Culture Techniques ,Pharmacokinetics ,In vivo ,medicine ,Animals ,Humans ,General Immunology and Microbiology ,Gene Expression Profiling ,Computational Biology ,Transporter ,Cell Biology ,drug metabolism ,030104 developmental biology ,Gene Expression Regulation ,Duodenum ,Caco-2 Cells ,Transcriptome ,Drug metabolism - Abstract
Induction of intestinal drug metabolizing enzymes can complicate the development of new drugs, owing to the potential to cause drug-drug interactions (DDIs) leading to changes in pharmacokinetics, safety and efficacy. The development of a human-relevant model of the adult intestine that accurately predicts CYP450 induction could help address this challenge as species differences preclude extrapolation from animals. Here, we combined organoids and Organs-on-Chips technology to create a human Duodenum Intestine-Chip that emulates intestinal tissue architecture and functions, that are relevant for the study of drug transport, metabolism, and DDI. Duodenum Intestine-Chip demonstrates the polarized cell architecture, intestinal barrier function, presence of specialized cell subpopulations, and in vivo relevant expression, localization, and function of major intestinal drug transporters. Notably, in comparison to Caco-2, it displays improved CYP3A4 expression and induction capability. This model could enable improved in vitro to in vivo extrapolation for better predictions of human pharmacokinetics and risk of DDIs. more...
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- 2020
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19. Author response: Duodenum Intestine-Chip for preclinical drug assessment in a human relevant model
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Geraldine A. Hamilton, Laxmi Sunuwar, Carolina Lucchesi, Katia Karalis, Raymond Luc, Mark Donowitz, Josiah Sliz, Magdalena Kasendra, Jenifer Obrigewitch, Dimitris V. Manatakis, Jianyi Yin, Kyung-Jin Jang, Gauri Kulkarni, and Athanasia Apostolou more...
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Drug ,medicine.anatomical_structure ,business.industry ,media_common.quotation_subject ,Duodenum ,Medicine ,Pharmacology ,business ,media_common - Published
- 2019
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20. A novel organ-chip system emulates three-dimensional architecture of the human epithelia and the mechanical forces acting on it
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Antonio Varone, Carolina Lucchesi, Katia Karalis, Christopher David Hinojosa, Riccardo Barrile, Norman Wen, Josiah Sliz, Achille Gravanis, Lian Leng, Justin Nguyen, and Geraldine A. Hamilton
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Materials science ,Interface (computing) ,Microfluidics ,Biophysics ,Bioengineering ,02 engineering and technology ,Biomaterials ,03 medical and health sciences ,Three dimensional architecture ,Lab-On-A-Chip Devices ,Humans ,Fluidics ,Endothelium ,030304 developmental biology ,0303 health sciences ,Microchannel ,Endothelial Cells ,021001 nanoscience & nanotechnology ,Chip ,Mechanics of Materials ,Ceramics and Composites ,Microtechnology ,0210 nano-technology ,Microfabrication ,Biomedical engineering ,Micropatterning - Abstract
Successful translation of in vivo experimental data to human patients is an unmet need and a bottleneck in the development of effective therapeutics. Organ-on-Chip technology aims to address this need by leveraging recent significant advancements in microfabrication and biomaterials, which enable modeling of organs and their functionality. These microengineered chips offer researchers the possibility to recreate critical elements of native tissue architecture such as in vivo relevant tissue-tissue interface, air-liquid interface, and mechanical forces, including mechanical stretch and fluidic shear stress, which are crucial to recapitulate tissue level functions. Here, we present the development of a new, comprehensive 3D cell-culture system, where we combined our proprietary Organ-Chip technology with the advantages offered by three-dimensional organotypic culture. Leveraging microfabrication techniques, we engineered a flexible chip that consists of a chamber containing an organotypic epithelium, surrounded by two vacuum channels that can be actuated to stretch the hydrogel throughout its thickness. Furthermore, the ceiling of this chamber is a removable lid with a built-in microchannel that can be perfused with liquid or air and removed as needed for direct access to the tissue. The bottom part of this chamber is made from a porous flexible membrane which allows diffusive mass transport to and from the microfluidic channel positioned below the membrane. This additional microfluidic channel can be coated with endothelial cells to emulate a blood vessel and recapitulate endothelial interactions. Our results show that the Open-Top Chip design successfully addresses common challenges associated with the Organs-on-Chip technology, including the capability to incorporate a tissue-specific extracellular matrix gel seeded with primary stromal cells, to reproduce the architectural complexity of tissues by micropatterning the gel, and to extract the gel for H&E staining. We also provide proof-of-concept data on the feasibility of using the system with primary human skin and alveolar epithelial cells. more...
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- 2021
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21. Human iPSC-Derived Blood-Brain Barrier Chips Enable Disease Modeling and Personalized Medicine Applications
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Jennifer E. Van Eyk, Magdalena Kasendra, Michael J. Workman, Matthew Rahnama, Zhaohui Chen, Weston R. Spivia, Jordan Kerns, Carolina Lucchesi, Sonalee Barthakur, Gad D. Vatine, Clive N. Svendsen, Samuel Sances, Bianca K. Barriga, Norman Wen, and Riccardo Barrile more...
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Brain vasculature ,Induced Pluripotent Stem Cells ,Microfluidics ,Drug Evaluation, Preclinical ,Bioengineering ,Disease ,Biology ,Blood–brain barrier ,Vascular lumen ,Capillary Permeability ,03 medical and health sciences ,Organ Culture Techniques ,0302 clinical medicine ,Genetics ,medicine ,Humans ,Precision Medicine ,Induced pluripotent stem cell ,Cells, Cultured ,030304 developmental biology ,Whole blood ,Neurons ,0303 health sciences ,business.industry ,Brain ,Cell Differentiation ,Transporter ,Cell Biology ,medicine.anatomical_structure ,Blood-Brain Barrier ,Astrocytes ,Molecular Medicine ,Endothelium, Vascular ,Personalized medicine ,business ,Neuroscience ,030217 neurology & neurosurgery - Abstract
Summary The blood-brain barrier (BBB) tightly regulates the entry of solutes from blood into the brain and is disrupted in several neurological diseases. Using Organ-Chip technology, we created an entirely human BBB-Chip with induced pluripotent stem cell (iPSC)-derived brain microvascular endothelial-like cells (iBMECs), astrocytes, and neurons. The iBMECs formed a tight monolayer that expressed markers specific to brain vasculature. The BBB-Chip exhibited physiologically relevant transendothelial electrical resistance and accurately predicted blood-to-brain permeability of pharmacologics. Upon perfusing the vascular lumen with whole blood, the microengineered capillary wall protected neural cells from plasma-induced toxicity. Patient-derived iPSCs from individuals with neurological diseases predicted disease-specific lack of transporters and disruption of barrier integrity. By combining Organ-Chip technology and human iPSC-derived tissue, we have created a neurovascular unit that recapitulates complex BBB functions, provides a platform for modeling inheritable neurological disorders, and advances drug screening, as well as personalized medicine. more...
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- 2019
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22. Benefits of oxygen and nitrogen plasma treatment in Vero cell affinity to poly(lactide-co-glycolide acid)
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Eliana Aparecida de Rezende Duek, Carolina Lucchesi, Betina M. P. Ferreira, Andrea Rodrigues Esposito, and Camila Mika Kamikawa
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Materials science ,Cell growth ,Mechanical Engineering ,Nanotechnology ,Adhesion ,Condensed Matter Physics ,Contact angle ,PLGA ,chemistry.chemical_compound ,Membrane ,chemistry ,Mechanics of Materials ,Biophysics ,General Materials Science ,Wetting ,Surface charge ,Cell adhesion - Abstract
Cell adhesion on materials surface is critical because this phenomenon occurs before other events, as cell spreading, cell migration and cell differentiation. it is commonly accepted that the adhesion of cells on solid substrate is influenced by several substratum surface properties, such as wettability, surface charge, roughness and topography. plasma technique is a convenient method for modifying surface properties of materials without affecting physical properties. in this study, poly(lactide-co-glycolide), plga, membranes were modified by oxygen and nitrogen plasma to improve polymer hydrophilicity and verify their effect on vero cells culture. the plga membranes, which were characterized by sem and contact angle, showed increased surface rugosity and narrower contact angles. cell adhesion, cytotoxicity assay, sem and cytochemistry analysis showed that plasma treatment was beneficial to cell growth by improving cell-polymer interaction. Cell adhesion on materials surface is critical because this phenomenon occurs before other events, as cell spreading, cell migration and cell differentiation. It is commonly accepted that the adhesion of cells on solid substrate is influenced by several substratum surface properties, such as wettability, surface charge, roughness and topography. Plasma technique is a convenient method for modifying surface properties of materials without affecting physical properties. In this study, poly(lactide-co-glycolide), PLGA, membranes were modified by oxygen and nitrogen plasma to improve polymer hydrophilicity and verify their effect on Vero cells culture. The PLGA membranes, which were characterized by SEM and contact angle, showed increased surface rugosity and narrower contact angles. Cell adhesion, cytotoxicity assay, SEM and cytochemistry analysis showed that plasma treatment was beneficial to cell growth by improving cell-polymer interaction. more...
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- 2013
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23. Human small airway-on-a-chip: A novel microphysiological system to model lung inflammation, accelerate drug development and enable inhalational toxico-analysis
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Rachelle Prantil-Baun, Mariko Hirano-Kobayashi, Carolina Lucchesi, Geraldine A. Hamilton, Remi Villenave, Youngjae Choe, Janna Nawroth, Donald E. Ingber, Kambez H. Benam, Richard M. Novak, James C. Weaver, Anthony Bahinski, Cedric Hubeau, and Thomas C. Ferrante more...
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COPD ,Lung ,business.industry ,medicine.medical_treatment ,Inflammation ,02 engineering and technology ,Disease ,021001 nanoscience & nanotechnology ,medicine.disease ,medicine.disease_cause ,030226 pharmacology & pharmacy ,respiratory tract diseases ,03 medical and health sciences ,0302 clinical medicine ,Cytokine ,medicine.anatomical_structure ,Drug development ,Immunology ,medicine ,medicine.symptom ,0210 nano-technology ,Airway ,business ,Oxidative stress - Abstract
Development of new therapeutics for lung inflammatory and infectious diseases, and advancement in our understanding of inhalational toxico-pathology have been hindered by challenges to study organ-level complexities of human lung in vitro . Here, we applied a microengineering technological approach known as 9organ-on-chip9 to create a human lung small airway-on-a-chip that supports full differentiation of a pseudostratified mucociliary bronchiolar epithelium from normal or diseased donors underlined by a functional microvascular endothelium. Small airway chips lined with chronic obstructive pulmonary disease (COPD) epithelia recapitulated features of the disease including selective cytokine hypersecretion, increased neutrophil recruitment, and clinical exacerbations by exposure to pathogens. Using this robust in vitro method, it was possible to detect synergistic tissue-tissue communication, identify new biomarkers of disease exacerbation, and measure responses to anti-inflammatory compounds that inhibit cytokine-induced recruitment of circulating neutrophils. Importantly, by connecting the small airway chip to a custom-designed electromechanical instrument that 9breathes9 whole cigarette smoke in and out of the chip microchannels, we successfully recreated smoke-induced oxidative stress, identified new ciliary micropathologies, and discovered unique COPD-specific molecular signatures. Moreover, this platform revealed a subtle ciliary damage triggered by acute exposure to electronic cigarette. Thus, the human small airway-on-a-chip offers a powerful complement to animal models for studying human lung pathophysiology. more...
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- 2016
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24. The Influence of the PCL-T Concentration in PLDLA Membrane
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Marcia Adriana Tomaz Duarte, Carolina Lucchesi, E. A. R. Duek, Larissa Coppini, and Marcelo Macedo
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Materials science ,Mechanical Engineering ,technology, industry, and agriculture ,Plasticizer ,Biomaterial ,macromolecular substances ,Condensed Matter Physics ,Lactic acid ,Solvent ,chemistry.chemical_compound ,Membrane ,chemistry ,Chemical engineering ,Mechanics of Materials ,Copolymer ,General Materials Science ,Composite material ,Cell adhesion ,Caprolactone - Abstract
The use of bioabsorbable polymers in biomedical application has increased greatly in recent years due to their good compatibility and bio-reabsorption. To obtain a polymeric material suitable for medical applications, the obtention and characterization of copolymer poly (L-co-DL lactic acid) (PLDLA) with the addition of poly (caprolactone triol) (PCL-T) was studied. PLDLA/PCL-T membranes were prepared by solvent casting in the 100/0, 90/10 and 70/30 (w/w) compositions. The membranes were characterized by Atomic Force Microscopy (AFM), Cellular Adhesion and Energy Dispersed Spectroscopy (EDS). Using MFA it was observed that an addition of PCL-T contributes to rough morphology and greater porosity. Increased cell adhesion on 90/10 and 70/30 membranes compared to 100/0 composition and controls was observed. From these results, it was observed that PCL-T improved cellular adhesion of the PLDLA membrane when compared to membranes without PCL-T. The PLDLA/PCL-T membrane is indicated for use in medical devices which do not require long implantation time, such as support for cell culture, dressings for skin ulceration and guided regeneration in periodontics. more...
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- 2012
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25. Cytocompatibility of poly(p-dioxanone)/ poly(hydroxybutic) (PPD/PHB) blends to cartilage tissue engineering
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Ana Paula Testa Pezzin, Andrea Rodrigues Esposito, Carolina Lucchesi, Luciana Prazeres, and E. A. R. Duek
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Blenda ,fibrochondrocytes ,Chemistry ,Organic Chemistry ,poli(p-dioxanona) ,fibrocondrócitos ,poly(hydroxybutic) ,Chemical Engineering (miscellaneous) ,Poly-p-dioxanone ,Blend ,poli(hidroxi butirato) ,Molecular biology ,poly(p-dioxanone) - Abstract
Buscando estratégias que repercutam na melhoria da interação entre materiais poliméricos biorreabsorvíveis e o crescimento celular, o presente estudo in vitro teve como objetivo estudar a influência de blendas de PPD/PHB na adesão celular e crescimento de fibrocondrócitos obtidos a partir de cultura primária. As blendas de PPD/PHB foram preparadas pelo método de evaporação de solvente nas composições 100/0, 60/40 e 50/50 e caracterizadas por microscopia eletrônica de varredura (MEV). Observações ultra-estruturais mostraram alterações na morfologia celular, sugerindo que os fibrocondrócitos podem responder a alterações no substrato alterando seu perfil fenotípico. As análises com MTT demonstraram que as blendas não apresentaram citotoxicidade e permitiram a adesão e proliferação dos fibrocondrócitos sobre os substratos em todas as suas composições. O ensaio colorimétrico com Sirius Red evidenciou a capacidade de manutenção da síntese de matriz extracelular colágena sobre as amostras, concluindo-se que as blendas de PPD/PHB podem ser indicadas para o cultivo celular. In order of seek strategies to improve the interaction between bioreabsorbable polymer materials and cellular growth, this work aimed at evaluating in vitro the influence of PPD/PHB blends on cell adhesion and fibrochondrocytes growth. Fibrochondrocytes cells were obtained by primary extraction from enzymatic digestion methods. The PPD/PHB blends were prepared by casting with 100/0, 60/40 and 50/50 compositions, and were characterized by scanning electron microscopy (SEM). After 6, 48, 120 and 168 hours in culture, ultrastructural observations showed changes in cell morphology, suggesting that the fibrochondrocytes can respond to substrate modifications, changing their phenotypic profile. The MTT analyses showed that the blends did not present cytotoxicity and allowed fibrochondrocytes adhesion and proliferation on the membranes in all compositions. The colorimetric Sirius Red test revealed the capability of extracellular matrix synthesis on the blends, from which one can conclude that the PPD/PHB blends are not cytotoxic and can be indicated for cell culture. more...
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- 2010
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26. Cell culture on PCL/PLGA blends
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Carolina Lucchesi, Samuel H. Barbanti, Eliana Aparecida de Rezende Duek, and Paulo Pinto Joazeiro
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Materials science ,Polymers and Plastics ,Biocompatibility ,Biomaterial ,General Chemistry ,Bone tissue ,Surfaces, Coatings and Films ,PLGA ,chemistry.chemical_compound ,medicine.anatomical_structure ,Chemical engineering ,Tissue engineering ,chemistry ,Cell culture ,Polycaprolactone ,Polymer chemistry ,Materials Chemistry ,medicine ,Cell adhesion - Abstract
Bioresorbable polymers have been studied as support for cell culture in the tissue engineering area. Osteoblastic cells were cultivated on poly(e-caprolactone), poly(lactic acid-co-glycolic acid), and (70/30), (50/50), and (30/70) blends. Cytotoxicity and cell adhesion assays and scanning electronic microscopy studies were described. The cells presented significant growth on the blends, showing no cytotoxic response. Results indicated that these blends are promising as devices for bone tissue applications. © 2009 Wiley Periodicals, Inc. J Appl Polym Sci, 2010 more...
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- 2010
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27. Increased response of Vero cells to PHBV matrices treated by plasma
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Carolina Lucchesi, Arnaldo R. Santos, Betina M. P. Ferreira, Paulo Pinto Joazeiro, and E. A. R. Duek
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Materials science ,Nitrogen ,Surface Properties ,Polyesters ,Biomedical Engineering ,Biophysics ,Biocompatible Materials ,Bioengineering ,Biomaterials ,Contact angle ,Tissue engineering ,Chlorocebus aethiops ,Polymer chemistry ,Cell Adhesion ,Animals ,Vero Cells ,chemistry.chemical_classification ,Tissue Engineering ,Tissue Scaffolds ,Substrate (chemistry) ,Polymer ,Adhesion ,Oxygen ,Polyester ,Membrane ,Chemical engineering ,chemistry ,Vero cell - Abstract
The copolymers poly(3-hydroxybutyric acid-co-3-hydroxyvaleric acid) (PHBV) are being intensely studied as a tissue engineering substrate. It is known that poly 3-hydroxybutyric acids (PHBs) and their copolymers are quite hydrophobic polyesters. Plasma-surface modification is an effective and economical surface treatment technique for many materials and of growing interest in biomedical engineering. In this study we investigate the advantages of oxygen and nitrogen plasma treatment to modify the PHBV surface to enable the acceleration of Vero cell adhesion and proliferation. PHBV was dissolved in methylene chloride at room temperature. The PHBV membranes were modified by oxygen or nitrogen-plasma treatments using a plasma generator. The membranes were sterilized by UV irradiation for 30 min and placed in 96-well plates. Vero cells were seeded onto the membranes and their proliferation onto the matrices was also determined by cytotoxicity and cell adhesion assay. After 2, 24, 48 and 120 h of incubation, growth of fibroblasts on matrices was observed by scanning electron microscopy (SEM). The analyses of the membranes indicated that the plasma treatment decreased the contact angle and increased the surface roughness; it also changed surface morphology, and consequently, enhanced the hydrophilic behavior of PHBV polymers. SEM analysis of Vero cells adhered to PHBV treated by plasma showed that the modified surface had allowed better cell attachment, spreading and growth than the untreated membrane. This combination of surface treatment and polymer chemistry is a valuable guide to prepare an appropriate surface for tissue engineering application. more...
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- 2007
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28. Study of vero cells/PLGA interaction after surface modification by oxygen plasma
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Andrea Rodrigues Esposito, E. A. R. Duek, Carolina Lucchesi, and Betina M. P. Ferreira
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Alternative methods ,Materials science ,Cell growth ,células vero ,PLGA ,General Physics and Astronomy ,Plasma treatment ,vero cells ,General Chemistry ,Adhesion ,Molecular biology ,Contact angle ,chemistry.chemical_compound ,Membrane ,chemistry ,Vero cell ,General Materials Science ,plasma ,Biomedical engineering - Abstract
A aplicação de polímeros bioreabsorvíveis como suporte para cultura de células é um método alternativo para o tratamento de lesões e perdas teciduais. A modificação da superfície desses polímeros por plasma é uma técnica efetiva e econômica para torná-los mais hidrofílicos e melhorar a adesão celular. O objetivo deste trabalho foi estudar as interações entre células Vero e suportes de PLGA previamente tratados por plasma de oxigênio, com o objetivo de aumentar a hidrofilicidade da superfície desses materiais. As amostras foram caracterizadas através das análises de ângulo de contato, MEV e citoquímica. O tratamento por plasma melhorou a adesão e a proliferação celular, em relação às membranas sem tratamento. The application of bioreabsorbable polymers as support for cells culture is an alternative method to treat lesions and loss of tissues. The surface modifications of these polymers by plasma is an effective and economical technique to make it more hydrophilic. The aim of this work was to study the interactions between Vero cells and PLGA supports previously treated by oxygen plasma, to increase the surface hydrofilicity. The samples were characterized by contact angle, MEV and citochemical analysis. The plasma treatment increased the hydrofilicity, improving the adhesion and cell proliferation, in comparison with the membranes without treatment. more...
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- 2007
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29. Cell adhesion to plasma-coated PVC
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Steven F. Durrant, Francine S. de Moraes, Eliana Aparecida de Rezende Duek, Eduardo S. de Souza, Wido H. Schreiner, Elidiane Cipriano Rangel, Nilson Cristino da Cruz, Carolina Lucchesi, Universidade Estadual Paulista (Unesp), Pontificia Univ Catolica, and Univ Fed Parana more...
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Materials science ,Article Subject ,Surface Properties ,lcsh:Medicine ,chemistry.chemical_element ,lcsh:Technology ,General Biochemistry, Genetics and Molecular Biology ,chemistry.chemical_compound ,X-ray photoelectron spectroscopy ,Coated Materials, Biocompatible ,Polymer chemistry ,Chlorocebus aethiops ,Cell Adhesion ,Animals ,lcsh:Science ,Cell adhesion ,Polyvinyl Chloride ,Vero Cells ,Deposition (law) ,General Environmental Science ,chemistry.chemical_classification ,lcsh:T ,lcsh:R ,General Medicine ,Adhesion ,Polymer ,Surface energy ,Polyvinyl chloride ,chemistry ,lcsh:Q ,Carbon ,Nuclear chemistry ,Research Article - Abstract
Made available in DSpace on 2015-03-18T15:55:35Z (GMT). No. of bitstreams: 0 Previous issue date: 2014-01-01Bitstream added on 2015-03-18T16:28:41Z : No. of bitstreams: 1 WOS000343426600001.epub: 1842935 bytes, checksum: bd7fc1f06020d4e1bc2b6d57366c4faf (MD5)Bitstream added on 2015-03-18T16:28:41Z : No. of bitstreams: 2 WOS000343426600001.epub: 1842935 bytes, checksum: bd7fc1f06020d4e1bc2b6d57366c4faf (MD5) WOS000343426600001.pdf: 7848804 bytes, checksum: 2d00acfe1706b61b21fc0f6c6cb6a001 (MD5) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) To produce environments suitable for cell culture, thin polymer films were deposited onto commercial PVC plates from radiofrequency acetylene-argon plasmas. The proportion of argon in the plasmas, P-Ar, was varied from 5.3 to 65.8%. The adhesion and growth of Vero cells on the coated surfaces were examined for different incubation times. Cytotoxicity tests were performed using spectroscopic methods. Carbon, O, and N were detected in all the samples using XPS. Roughness remained almost unchanged in the samples prepared with 5.3 and 28.9% but tended to increase for the films deposited with P-Ar between 28.9 and 55.3%. Surface free energy increased with increasing P-Ar, except for the sample prepared at 28.9% of Ar, which presented the least reactive surface. Cells proliferated on all the samples, including the bare PVC. Independently of the deposition condition there was no evidence of cytotoxicity, indicating the viability of such coatings for designing biocompatible devices. Univ Estadual Paulista UNESP, Lab Plasmas Tecnol, BR-18087180 Sorocaba, SP, Brazil Pontificia Univ Catolica, Dept Ciencias Fisiol, Lab Biomat, BR-18030095 Sorocaba, SP, Brazil Univ Fed Parana, Dept Fis, Lab Superficies & Interfaces, BR-81531990 Curitiba, Parana, Brazil Univ Estadual Paulista UNESP, Lab Plasmas Tecnol, BR-18087180 Sorocaba, SP, Brazil FAPESP: 05/03420-4 FAPESP: 11/21345-0 more...
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- 2014
30. Vero cells culture on poly poli(hydroxybutyrate-co-hydroxyvalerate) (phbv) membranes treated by gaseous plasma
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Carolina Lucchesi, Joazeiro, Paulo Pinto, 1953, Moreira, Patricia da Luz, Santos Junior, Arnaldo Rodrigues dos, Universidade Estadual de Campinas. Instituto de Biologia, Programa de Pós-Graduação em Biologia Celular e Estrutural, and UNIVERSIDADE ESTADUAL DE CAMPINAS more...
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Biomaterials ,Celulas vero ,Blood plasma ,poli(hidroxibutirato-co-hidroxivalerato) ,Biomateriais ,Plasma sanguíneo ,poly (hydroxybutyrate-co-hydroxyvalerate) (PHBV) - Abstract
Orientador: Paulo Pinto Joazeiro Dissertação (mestrado) - Universidade Estadual de Campinas, Instituto de Biologia Resumo: O copolímero Poli(hidroxibutirato-co-hidroxivalerato) (PHBV) tem sido intensamente estudado como substrato para a engenharia de tecidos, sendo conhecido como um poliéster hidrofóbico. A modificação da superfície por plasma é uma técnica efetiva e econômica para os materiais e tem ganhado crescente interesse da engenharia biomédica, por melhorar a biocompatibilidade da superfície. Neste estudo, avaliou-se as vantagens da modificação da superfície de membranas de PHBV tratadas por plasma de Oxigênio e Nitrogênio a fim de acelerar o processo de adesão e proliferação celular. O PHBV foi dissolvido em c1oreto de metileno à temperatura ambiente. Membranas de PHBV foram submetidas ao tratamento de plasma de Oxigênio e Nitrogênio, através de um gerador de plasma. As membranas foram esterilizadas por radiação UV por 30 min e colocadas em placas de 96 poços. Células Vero foram semeadas sobre as membranas, sendo determinada a proliferação celular sobre as matrizes, a citotoxicidade e adesão celular. Após 2, 24,48 e 120h de incubação, o crescimento e proliferação dos fibroblastos foram observados por microscopia eletrônica de varredura (MEV). As análises das membranas indicaram que o tratamento por plasma aumentou o ângulo de contato e a rugosidade, alterando a morfologia da superfície, e conseqüentemente, melhorou ocomportamento hidrofílico do polímero. A microscopia eletrônica de varredura das células Vero mostrou que as modificações da superfície proporcionaram melhor adesão, espalhamento e proliferação celular. O tratamento da superfície do polímero somado às suas propriedades químicas é um caminho para obtenção de estruturas aplicáveis a engenharia de tecido Abstract: The copolymers poly(3-hydroxybutyric acid-co-3-hydroxyvaleric acid) (PHBV) are being intensely studied as a tissue engineering substrate. It is know that Poly(hydroxybutic) (PHB) and their copolymers are quite hydrophobic polyesters. Plasma-surface modification is an effective and economical surface treatment technique for many materiaIs and of growing interest in biomedical engineering. In this study we investigate the advantages of oxygen and nitrogen plasma treatment to modify the PHBV surface to enable the acceleration of Vero cell adhesion and proliferation. PHBV was dissolved in methylene chloride at room temperature. The PHBV membranes were modified by oxygen or nitrogen-plasma treatments using a plasma generator. The membranes were sterilized by UV irradiation for 30 min and placed in 96-well plates. Vero cells were seeded onto the membranes and their proliferation onto the matrices was also determined by cytotoxicity and cell adhesion assay. After 2, 24, 48 and 120h of incubation, growth of fibroblasts on matrices was observed by scanning electron microscopy (SEM). The analyses of the membranes indicated that the plasma treatment increased the contact angle and their roughness, it also changed the surface morphology, and consequently, enhanced the hydrophilic behavior of PHBV polymers. Scanning electron microscopy analysis of Vero cell adhered to plasma treated PHBV showed that the modified surface had allowed better cell attachment, spreading and growth than the untreated membrane. This combination of surface treatment and polymer chemistry is a valuable guide to prepare appropriated surface for tissue engineering application Mestrado Histologia Mestre em Biologia Celular e Estrutural more...
- Published
- 2006
31. On the design of clone-based haplotyping
- Author
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Susan M. Byrne, Christine Lo, Vineet Bafna, Carolina Lucchesi, Kimberly Robasky, George M. Church, John Aach, Je-Hyuk Lee, Kun Zhang, and Rui Liu
- Subjects
Chromosomes, Artificial, Bacterial ,Computational biology ,Biology ,Polymorphism, Single Nucleotide ,Genome ,Contig Mapping ,03 medical and health sciences ,Disease susceptibility ,0302 clinical medicine ,HLA Antigens ,Humans ,Cloning, Molecular ,030304 developmental biology ,Genetics ,0303 health sciences ,Genome, Human ,Research ,Haplotype ,Chromosome ,Sequence Analysis, DNA ,Human genetics ,Molecular Typing ,Haplotypes ,Human genome ,Algorithms ,030217 neurology & neurosurgery ,Personal genomics - Abstract
Background Haplotypes are important for assessing genealogy and disease susceptibility of individual genomes, but are difficult to obtain with routine sequencing approaches. Experimental haplotype reconstruction based on assembling fragments of individual chromosomes is promising, but with variable yields due to incompletely understood parameter choices. Results We parameterize the clone-based haplotyping problem in order to provide theoretical and empirical assessments of the impact of different parameters on haplotype assembly. We confirm the intuition that long clones help link together heterozygous variants and thus improve haplotype length. Furthermore, given the length of the clones, we address how to choose the other parameters, including number of pools, clone coverage and sequencing coverage, so as to maximize haplotype length. We model the problem theoretically and show empirically the benefits of using larger clones with moderate number of pools and sequencing coverage. In particular, using 140 kb BAC clones, we construct haplotypes for a personal genome and assemble haplotypes with N50 values greater than 2.6 Mb. These assembled haplotypes are longer and at least as accurate as haplotypes of existing clone-based strategies, whether in vivo or in vitro. Conclusions Our results provide practical guidelines for the development and design of clone-based methods to achieve long range, high-resolution and accurate haplotypes. more...
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