Dissertations / Theses on the topic 'In vitro platform'
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Harrison, Olivia Jane. "Integrated platform to assay melanoblast development in vitro." Thesis, University of Edinburgh, 2018. http://hdl.handle.net/1842/31164.
Jeon, Jessie Sungyun. "In vitro study of cancer cell extravasation in microfluidic platform." Thesis, Massachusetts Institute of Technology, 2014. http://hdl.handle.net/1721.1/87976.
Cataloged from PDF version of thesis.
Includes bibliographical references.
Cancer metastases arise from the cancer cells that disseminate from the primary tumor, intravasate into the vascular system and eventually transmigrate across the endothelium into to a secondary site through a process of extravasation. Microfluidic systems have a major advantage in studying cancer extravasation since they can mimic aspects of the 3D in vivo situation in a controlled environment while simultaneously providing in situ imaging capabilities for visualization, thereby enabling quantification of cell-cell and cell-matrix interactions. Moreover, microfluidics enable parametric study of multiple factors in controlled and repeatable conditions. This thesis describes novel 3D microfluidic models to mimic the tumor microenvironment and vasculature during cancer cell extravasation in order to investigate the critical steps of extravasation. First, a general non-organ-specific cancer cell extravasation model is developed in which the endothelial cells that cover the walls of the microfluidic channel represent the vessel endothelium, and the entire extravasation process including tumor cell adhesion to the endothelium and subsequent transmigration can be observed. A second model is then introduced to mimic organ-specific extravasation and investigate the preference of certain types of cancer to target specific organs for metastass. The improved model was used to study the specificity of human breast cancer metastases to bone, by recreating a vascularized bone-mimicking microenvironment. The tri-culture system allowed us to study the transendothelial migration of highly metastatic breast cancer cells and to monitor their behavior within the bone-like matrix. Next, functional microvascular networks were generated in the microfluidic system through vasculogenesis with addition of mural cells and pro-angiogenic factors to better replicate the normal physiological vasculature of the remote site for metastasis. Lastly, the vasculogenesis approach was combined with the bone-mimicking model to develop a functional osteo-cell conditioned vasculature model to study physiologically relevant extravasation in a bone-like microenvironment. In addition to the quantification of extravasation rates and subsequent tumor cell migration into the model tissue, the vascular networks were characterized by measuring permeability, and immunostaining of proteins secreted by osteo-cell and mural cell markers confirmed the creation of microenvironments and the presence of multiple cell types within the matrix. This study provides novel 3D in vitro quantitative data on cancer cell extravasation and micrometastasis of breast cancer cells within a bone-mimicking microenvironment. The developed microfluidic system represents an advanced in vitro model to study complex biological phenomena such as extravasation involving functional microvascular networks under organ-specific conditions and demonstrates the potential value of microfluidic technologies to better understand cancer biology and screen for new therapeutics.
by Jessie Sungyun Jeon.
Ph. D.
Chen, Michelle B. (Michelle Berkeley). "Tumor cell extravasation in an in vitro microvascular network platform." Thesis, Massachusetts Institute of Technology, 2014. http://hdl.handle.net/1721.1/93857.
Cataloged from PDF version of thesis.
Includes bibliographical references at the end of each chapter.
A deeper understanding of the mechanisms of tumor cell extravasation is essential in creating therapies that target this crucial step in cancer metastasis. Extravasation assays exist, but with limitations; data from in vivo models are frequently inferred from low-resolution end-point assays while most in vitro platforms are limited in their physiological relevance of the tumor microenvironment. To address this need, we developed a microfluidic platform to study tumor cell extravasation from in vitro microvascular networks formed via vasculogenesis. Various techniques to yield optimal networks were assessed in order to achieve an appropriate balance between vascular growth, remodeling and stabilization. These include the application of various soluble biochemical factors and both paracrine and juxtacrine co-culture with stromal cells. We demonstrate that out of all methods attempted, paracrine non-contact co-culture with human lung fibroblasts yield the most interconnected and stable networks. Vasculatures developed exhibit tight endothelial cell-cell junctions, basement membrane deposition and physiological values of vessel permeability. Employing our assay, we demonstrate impaired endothelial barrier function and increased extravasation efficiency with inflammatory cytokine stimulation, as well as positive correlations between the metastatic potentials of tumor cells lines and their extravasation capabilities. High-resolution time-lapse microscopy reveals the highly dynamic nature of extravasation events, beginning with thin tumor cell protrusions across the endothelium followed by extrusion of the remainder of the cell body through the formation of sub nuclear sized openings in the endothelial barrier. No disruption to endothelial cell-cell junctions is discernible at 60X, or by changes in local barrier function after completion of transmigration. Using our platform, we also elucidate the extravasation patterns of different tumor cell subpopulations, including mechanically lodged cells, single arrested non-trapped cells, and tumor cell clusters. Our platform offers key advantages over existing in vitro extravasation models by enabling all of the following: (1) high temporal and spatial resolution of extravasation events, (2) the ability to perform parametric studies in a tightly controlled and high throughput microenvironment and (3) increased physiological relevance compared to 2D and 3D planar monolayer models. Findings from our platform result in a deeper understanding of tumor cell extravasation mechanisms and demonstrate our assay's potential to be employed for the discovery of factors that could inhibit this crucial step in metastasis.
by Michelle B. Chen.
S.M.
Kim, Jae Eung. "In Vitro Synthetic Biology Platform and Protein Engineering for Biorefinery." Diss., Virginia Tech, 2017. http://hdl.handle.net/10919/86645.
Ph. D.
MELLE, GIOVANNI. "Development of a Novel Platform for in vitro Electrophysiological Recording." Doctoral thesis, Università degli studi di Genova, 2020. http://hdl.handle.net/11567/1000590.
Rotolo, Jimmy A. "Ceramide-mediated platform generation regulates apoptosis in vitro and in vivo /." Access full-text from WCMC:, 2007. http://proquest.umi.com/pqdweb?did=1428842781&sid=10&Fmt=2&clientId=8424&RQT=309&VName=PQD.
Petrucci, Teresa. "Building a platform for flexible and scalable testing of genetic editors." Doctoral thesis, Università di Siena, 2021. http://hdl.handle.net/11365/1143160.
Nordh, Nicki. "Development of a cell cultureplatform in PDMS : Microfluidic systems for in vitro productionof platelets." Thesis, Uppsala universitet, Mikrosystemteknik, 2015. http://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-261711.
Kim, Kihwan. "MULTICEULLULAR TUMOR HEMI-SPHEROID: A NOVEL IN VITRO 3D MODEL PLATFORM FOR ACCELERATED DRUG DEVELOPMENT." Case Western Reserve University School of Graduate Studies / OhioLINK, 2017. http://rave.ohiolink.edu/etdc/view?acc_num=case1481900120946458.
FICULLE, ELENA. "DEVELOPMENT OF AN IN-VITRO HUMANIZED MICROFLUIDIC PLATFORM TO STUDY NEURONAL TAU AGGREGATION AND PROPAGATION." Doctoral thesis, Università degli Studi di Milano, 2021. http://hdl.handle.net/2434/807615.
Background: Alzheimer’s disease (AD) is the most common cause of dementia, characterized by the presence of extracellular -amyloid plaques and intracellular neurofibrillary tangles (NFTs) composed of aggregated and hyperphosphorylated tau. Since it has been shown that tau aggregates correlate with cognitive decline much better than -amyloid formations, it is important to understand how tau can spread in the brain. Moreover, the spatiotemporal spread of tau observed during clinical manifestation suggests that it propagates along the axonal network between synaptically connected neurons. On these bases, some early-phase Clinical Trials are aiming to target tau during transcellular spreading in order to prevent its internalization by recipient neurons, using both compounds and antibodies. For the experimental evaluation of candidates before in vivo studies, there have been many attempts to replicate this network in vitro, most of which used microfluidic approaches; however, these experiments have often utilized parameters that may reduce the physiological relevance of the assay, such as by overexpressing tau, using fluorescent tags, or by introducing MAPT mutations. New methods that can improve these assays are required to help the screening of efficient and effective treatments. Aim of the work: The main purpose of this research project has been to establish a humanized, in vitro neuronal microfluidic platform to recapitulate and study tau aggregation and propagation in a qualitative and quantitative way. Microfluidic devices represent a miniaturized alternative tool to recapitulate tau spreading conditions, by enabling the culture of synaptically connected, but environmentally isolated, neuronal populations that can be seeded, thereby inducing endogenous tau aggregation and subsequent propagation. This model system could be ideal for testing the effect of potential tau therapeutics that modulate transneuronal tau propagation. Material & Methods: Before developing the microfluidic propagation assay, a rat cortical neuron (RCN) aggregation assay that uses seeding-competent material from human AD brains (hAD seed) to induce endogenous aggregation was validated. Subsequently, the same conditions were used to develop a RCN microfluidic assay that can show endogenous tau aggregation, and consequent propagation, using High Content Imaging (HCI) and a proprietary interactive computer program for image quantification. Finally, using a transgenic mouse line that expresses human MAPT, a humanized and miniaturized version of the assay has been developed in order to have a physiologically relevant, medium-throughput platform to test tau therapies. Results: After a phase of optimization, it has been shown that hAD seed induces endogenous rodent tau aggregation and transneuronal propagation in a quantifiable manner in a microfluidic culture model. Moreover, this assay was statistically validated and further converted to a medium-throughput format allowing the user to handle 16 two-chamber devices simultaneously in the footprint of a standard 96 well plate. Furthermore, this assay was humanized in order to study hTau aggregation and propagation using primary neurons from a mouse model that expresses human tau only. It has been proved that Anle 138b, a literature small molecule that has been previously shown to impair protein aggregation, can block the transneuronal transfer of tau aggregates, suggesting that this novel system can be used to evaluate mechanisms of tau spreading and to find therapeutic interventions. Moreover, preliminary experiments have shown that the aggregation of endogenous tau induces not only an increase of neuronal excitability but also an activation of astrocytes which might also have a role in tau pathology. Conclusions: This work has been successfully developed a robust and quantitative microfluidic assay that can model an isolated mechanism of tau propagation. Using both RCNs and hTau mouse cortical neurons seeded with hAD seed, it was possible to quantify the formation and propagation of endogenous tau inclusions and demonstrate that a putative inhibitor of tau propagation is active in this assay. It was also shown that these models can be further employed for exploratory studies, such as monitoring the functional activity and connectivity of the neuronal cultures, as well as investigating the role of different cell types in tau aggregation and propagation. Most importantly, this manuscript exhibits the latent potential of microfluidic assays as screening platforms for the preclinical evaluation of tau propagation inhibitors.
Reichert, Verena Maria Charlotte. "Application of a human bone engineering platform to an in vitro and in vivo breast cancer metastasis model." Thesis, Queensland University of Technology, 2011. https://eprints.qut.edu.au/53212/1/Verena_Reichert_Thesis.pdf.
Carletti, Angelo. "Development of a machine learning algorithm for the automatic analysis of microscopy images in an in-vitro diagnostic platform." Master's thesis, Alma Mater Studiorum - Università di Bologna, 2021.
Börnicke, Carl Jonathan [Verfasser]. "In vitro Reconstitution and Characterization of Soluble Complexes of the Salmonella Type III Secretion System Sorting Platform / Carl Jonathan Börnicke." Hamburg : Staats- und Universitätsbibliothek Hamburg Carl von Ossietzky, 2020. http://d-nb.info/1227853734/34.
Canuto, Gisele André Baptista. "Avaliação metabolômica comparativa in vitro de fármaco candidato ao tratamento de leishmaniose." Universidade de São Paulo, 2016. http://www.teses.usp.br/teses/disponiveis/46/46136/tde-14092016-090130/.
Leishmaniasis is a neglected disease caused by Leishmania parasites, which occurs in more than 90 countries worldwide, affecting millions of people. The disease is treatable but not curable, and the commonly used therapeutic arsenal is limited and consists in the use of highly toxic drugs. The search for new therapies has been encouraged and the use of active compounds, isolated from natural products, such as plants, has been proven effective. For future use of these active compounds in the treatment, it is necessary to understand its mechanism of action. \"Omics\" sciences, specifically metabolomics, which is the comparative analysis of altered metabolites in a biological system after external intervention, has been widely applied for this purpose. The metabolomic approach is multidisciplinary and the analyzes are performed using modern analytical platforms, such as separation techniques coupled to mass spectrometry (MS). Data treatment are performed with advanced statistical tools and altered metabolites are correlated with metabolic pathways. In this work, the mechanism of action, in Leishmania infantum, of an active compound (methyldehydrodieugenol B), isolated from Nectandra leucantha plant, was evaluated by global metabolomics approach with multiplatform analysis (gas and reversed phase liquid chromatography, GC-MS and RPLC-MS, respectively). Optimizations of sample preparation, extraction and derivatization for GC-MS to obtain the maximum number of metabolites in the samples under consideration were performed. L. infantum promastigotes were grown in culture medium and after 72 h they were treated with metildihidrodieugenol B (at a concentration of 58.18 µg x mL-1); after 48 h of treatment, enzyme activity was quenched, cells washed and frozen for further analysis. Methanol:water (1:1) and 100% methanol are used as solvent extractor to assess intracellular metabolites for analysis by GC-MS and RPLC-MS, respectively. Derivatization with O-methoxyamine in pyridine (15 mg x mL-1) at room temperature for 90 min, followed by silylation with BSTFA + 1% TMCS for 30 min at 40°C was performed to make volatile and stable metabolites for analysis by GC- MS. The results of both analytical platforms showed significant differences between treated and non-treated samples with the active substance, and altered metabolites with statistical significance were correlated with Leishmania metabolism. Different amino acids, fatty acids, carbohydrates, and glycerolipids phospholipids were found, mostly decreased with treatment. Due to the complexity of the parasite metabolism and the great diversity of altered metabolites, a multi-target mechanism can be assigned to the methyldehydrodieugenol B. Changes in the metabolism of glycolysis and gluconeogenesis are highlighted, indicating changes in the cell energy sources. Changes in the lipid composition of the parasite plasma membrane were also observed, suggesting variations in its flexibility and fluidity. The results are preliminary, but very important to take the first step into elucidation of the action of this substance in the treatment of leishmaniasis, aiming the search for new pharmaceutical therapies.
Lee, Bill. "Preclinical antimicrobial drug discovery : development and evaluation of a platform for high-throughput screening in vitro and an immunocompromised animal model." Thesis, McGill University, 2006. http://digitool.Library.McGill.CA:80/R/?func=dbin-jump-full&object_id=100745.
Chung, Git Weng. "Development of an in vitro rat proximal tubule cell model as a platform for drug transporter and drug-drug interaction studies." Thesis, University of Newcastle upon Tyne, 2014. http://hdl.handle.net/10443/2755.
Allenby, Mark Colin, Athanasios Mantalaris, and Nicki Panoskaltsis. "Development of a bio-inspired in silico-in vitro platform: Towards personalised healthcare through optimisation of a bone-marrow mimicry bioreactor." Thesis, Imperial College London, 2017. https://eprints.qut.edu.au/199969/1/Allenby_MC_2017_PhD_Thesis.pdf.
BEZUKLADOVA, SVETLANA. "Development of an in vitro functional platform to discover new drugs for progressive multiple sclerosis and translation to preclinical animal models." Doctoral thesis, Università Vita-Salute San Raffaele, 2022. http://hdl.handle.net/20.500.11768/133063.
La sclerosi multipla (SM), una malattia infiammatoria autoimmune caratterizzata da alterazione della mielina e da danno assonale, è la principale causa di disabilità causata da neurodegenerazione progressiva nei giovani adulti. La maggior parte dei pazienti con SM ha un accumulo continuo di disabilità per tutta la durata della vita che porta alla SM progressiva (SMP), una malattia ancora incurabile. La sfida urgente è lo sviluppo di nuove terapie efficaci per le forme progressive di SM, che potrebbero rallentare o prevenire la neurodegenerazione in corso, favorendo meccanismi di neuroprotezione e di rimielinizzazione. Uno degli approcci per raggiungere questo obiettivo è il riposizionamento di farmaci già presenti sul mercato, e che prevede l'identificazione di nuovi bersagli per i suddetti medicinali allo scopo di trattare altre malattie. Oltre al riutilizzo dei farmaci, la predizione dei farmaci in silico offre un approccio complementare per la selezione di validi candidati farmacologici di successo. Per riprodurre meglio il tessuto biologico umano, le cellule staminali pluripotente indotte (CSPi) possono fornire uno strumento prezioso per la modellazione della malattia in vitro e una fonte cellulare quasi infinita, preservando il background genetico del donatore. Le colture di oligodendrociti e di cellule neuronali derivate da CSPi rappresentano uno strumento unico per modellare le patologie in modo personalizzato per il paziente e per valutare gli effetti pro-mielinizzanti e neuroprotettivi dei farmaci candidati. Lo scopo del progetto è condurre uno screening farmacologico completo e ben caratterizzato, per identificare in definitiva un set di composti con potenziale terapeutico per la SMP. Pertanto, è stato condotto uno screening in vitro su una libreria di farmaci candidati per riposizionamento, utilizzando colture primarie di cellule neuronali murine e da CSPi di derivazione umana. La libreria di farmaci da riposizionare in questo progetto è stata prodotta dallo strumento bioinformatico SPOKE, sviluppato dal laboratorio di S. Baranzini dell'UCSF. L'attività farmacologica di due farmaci selezionati è stata valutata in vivo nel modello preclinico di SM (EAS). Sono attualmente in corso l'identificazione del bersaglio e lo studio del meccanismo d'azione delle proprietà neuroprotettive/rimielinizzanti di questi composti. Inoltre, abbiamo caratterizzato i profili molecolari e i fenotipi funzionali dei neuroni derivati da CSPi da tre coppie gemelle discordanti per la SM al fine di ottenere ulteriori conoscenze sul potenziale impatto dei cambiamenti genetici ed epigenetici sulla suscettibilità del danno neuronale nella SM. Comprendendo meglio le cause alla base della progressione del danno nel sistema nervoso centrale (SNC) e, in particolare, della ridotta capacità di rigenerazione della mielina, lo sviluppo di nuovi trattamenti per i pazienti di sclerosi multipla progressiva sarà infine accelerato.
VISENTIN, CRISTINA. "Use of a technological platform to screening in vitro and in vivo anti-amyloidogenic drugs able to prevent early neurodegenerative process." Doctoral thesis, Università degli Studi di Milano-Bicocca, 2017. http://hdl.handle.net/10281/158278.
Amyloidoses are protein misfolding diseases caused by deposition of fibrillar proteins in target organs. Nowadays, most of them are still incurable and their relevance to public health system is growing, especially as a consequence of population aging. Spinocerebellar ataxia type 3 is a member of this group of pathologies and its causative agent is ataxin-3 (ATX3). This is consists of a globular N-terminal (JD), followed by a flexible tail carrying a poly-glutamine (polyQ) tract. An expanded polyQ tract triggers the aggregation. In this work, I have investigated the capability of tetracycline (Tetra), epigallocatechin-gallate (EGCG), epigallocatechin (EGC), gallic acid (GA) and trifluoroethanol (TFE) to interfere with ATX3 amyloid deposition. Tetra is an antibiotic recently re-evaluated as anti- amyloidogenic compound. EGCG, EGC and GA, which are natural polyphenols, are already known in literature for their anti-amyloidogenic effect; finally, TFE is an osmolyte that stabilizes secondary structure, preferentially α-helix. Data obtained by aggregation assay, spectroscopic analyses (NMR, FTIR) and morphologic characterisation clearly demonstrated Tetra capability of increasing ATX3 aggregates solubility, without a substantial remodelling of the internal structure. Nevertheless, this antibiotic reduced the toxicity of the oligomeric species and ameliorated ataxic C. elegans phenotype. On the contrary, the analysed polyphenols were capable to interfere with ATX3 aggregation but, instead of preventing, they accelerated the aggregation rate redirecting the process towards the formation of soluble, not toxic, off-pathway aggregates. All compounds were also active against the JD in isolation, but only the polyphenols were capable to bind the monomeric form. In particular, they overlapped specific aggregation-prone regions directly involved in the fibrillation. This could explain their capability of redirecting the aggregation pathway and the different mode of action with respect to Tetra. These polyphenols showed a remarkable reduction of ATX3-mediated cytotoxicity and mitigation of ataxic phenotype in C. elegans and E. coli models. However, the compounds displayed a different efficacy, whereby EGCG was the most and GA the least effective. All data strongly support the idea that GA is the minimal functional unit of EGCG. TFE did not show the capability of preventing aggregation; in fact, even at very low concentration it promotes a faster amyloid-like aggregation. Biophysical characterization of its effect on JD aggregation, instead, provided evidence that ATX3 aggregation proceeds along a new identified pathway by which protein misfolding follows protein aggregation. In fact, TFE induces the formation of a native-like state almost indistinguishable from fully native protein, but more aggregation prone.
Billington, Sarah Faye. "Development of in-vitro human and rat proximal tubule cell models as a platform for drug transporter and drug-drug interaction studies." Thesis, University of Newcastle upon Tyne, 2015. http://hdl.handle.net/10443/3031.
Kinfu, Birhanu Mekuaninte [Verfasser], and Wolfgang [Akademischer Betreuer] Streit. "Function-based searches for selected phosphotransferases and establishing in vitro transcription platform for cell-free metagenomics / Birhanu Mekuaninte Kinfu ; Betreuer: Wolfgang Streit." Hamburg : Staats- und Universitätsbibliothek Hamburg, 2018. http://d-nb.info/1171427328/34.
Körfer, Georgette [Verfasser], Ulrich [Akademischer Betreuer] Schwaneberg, and Lothar [Akademischer Betreuer] Elling. "Development of a flow cytometer-based in vitro compartmentalization screening platform for directed protein evolution / Georgette Dorothea Johanna Körfer ; Ulrich Schwaneberg, Lothar Elling." Aachen : Universitätsbibliothek der RWTH Aachen, 2016. http://d-nb.info/1159380074/34.
Wang, Eu Sheng. "Construction and molecular characterisation of an improved chloroplast transformation vector system as a versatile delivery and expression platform for in-vitro propagated Nicotiana benthamiana." Thesis, University of Nottingham, 2016. http://eprints.nottingham.ac.uk/30486/.
Kabulski, Jarod L. "Development of Au-immobilized P450 platform for exploring the effect of oligomer formation on P450-mediated metabolism for In vitro to In vivo drug metabolism predictions." Morgantown, W. Va. : [West Virginia University Libraries], 2010. http://hdl.handle.net/10450/10892.
Title from document title page. Document formatted into pages; contains xiv, 180 p. : ill. (some col.). Includes abstract. Includes bibliographical references.
Kirichuk, Oksana. "Avancées dans les études in vitro des interactions cellule-glycocalyx : développement d'une plateforme définie mécaniquement et biochimiquement." Electronic Thesis or Diss., Université Grenoble Alpes, 2023. http://www.theses.fr/2023GRALY084.
Cell adhesion to the blood vessel wall is a complex, highly regulated physiological process. Red blood cells must repel from the blood vessel wall to prevent blood clotting while immune cells can be recruited from the vascular system to migrate into surrounding tissues. Cell adhesion hinges on the critical role played by the glycocalyx, a soft gel-like layer coating the vascular wall. However, how glycocalyx mechanical (softness, thickness) and biochemical (the composition and the density of surface receptors) properties affect this regulation is still poorly understood. Our hypothesis is that selective cell adhesion requires an intricate interplay of mechanical and biochemical cues. Elucidating the physical and molecular mechanisms that underpin selective adhesion directly in real blood vessels is challenging owing to the complexity and lack of control in in vivo systems. In my research, I aimed to construct an in vitro molecular interaction platform to facilitate mechanistic analyses. The platform combines a molecularly-defined model of the glycocalyx with mimetics of white blood cells under flow. While developing such a platform posed challenges, it offers the advantage of precise control over the physical and biochemical parameters of both the glycocalyx mimetic and cell mimetics.The newly developed glycocalyx model includes several key ingredients with tightly controlled properties: a brush of hyaluronan (HA, an essential component of the endothelial glycocalyx) is combined with P-selectin (an adhesion molecule on the endothelial cell surface critical for the homing of leukocytes). Building on previous experience in my research group, I employed a silica-supported lipid bilayer (SLB) bearing a monolayer of streptavidin (SAv), that can bind biotinylated molecules via biotin-SAv bonds. I introduce here a control of the in-plane mobility of molecules anchored to the fluid lipid bilayer using glutaraldehyde (GTA) as a cross-linking agent for SAv. Controlled grafting densities of one-end biotinylated HA chains of various lengths then create brushes of different mechanical properties. I also present a new methodology for quantitatively tuning the grafting density of smaller biotinylated molecules, which is deployed here to control the grafting density of an ‘adapter protein’ for anchoring P-selectin. The new in vitro model of the glycocalyx thus affords control over the lateral mobility, the surface density and the orientation of two distinct functional molecules.The second key component of the newly developed platform consists of white blood cell mimetics, developed based on commercially available microbeads with the size of a cell and a SAv coating. I introduce a methodology for simultaneous grafting of two types of proteins onto the bead surface: biotinylated CD44 (a ligand expressed on leukocyte surfaces, interacting specifically with HA) and PSGL-1 (a ligand of P-selectin). Additionally, I present a method for controlling the surface density of each of these proteins.I use a combination of methods as monitoring and quality control tools of glycocalyx model formation and bead functionalization: quartz crystal microbalance with dissipation monitoring (QCM-D); spectroscopic ellipsometry (SE), reflection interference contrast microscopy (RICM); confocal microscopy with fluorescence recovery after photobleaching (FRAP) capabilities, and flow cytometry.This newly established platform provides a controlled environment for studying blood cell adhesion, effectively bridging the divide between cell-glycocalyx chemical interactions and the mechanical aspects of cell migration under flow, including attachment and repulsion from the vascular wall. This platform holds the potential for expansion to encompass other surface adhesion molecules or to integrate multiple adhesion molecules, to gradually advance from the bottom up our understanding of the mechanisms governing cell adhesion to blood vessels
Guan, Shan [Verfasser], and Carsten [Akademischer Betreuer] Rudolph. "Peptide-based platform enabling amphiphilic block copolymers to acquire in vitro transfection ability and more potent in vivo lung gene transfer for cystic fibrosis / Shan Guan ; Betreuer: Carsten Rudolph." München : Universitätsbibliothek der Ludwig-Maximilians-Universität, 2018. http://d-nb.info/1207270016/34.
Mosaad, Eman Mohamed Othman. "Three dimensional prostate cancer model systems." Thesis, Queensland University of Technology, 2018. https://eprints.qut.edu.au/118287/1/Eman%20Mohamed%20Othman_Mosaad_Thesis.pdf.
Júnior, Mário Roberto Moraes. "Formação in vitro de biofilme de Candida albicans em materiais usados no preenchimento dos acessos aos parafusos das próteses sobre implantes." Universidade do Estado do Rio de Janeiro, 2012. http://www.bdtd.uerj.br/tde_busca/arquivo.php?codArquivo=6132.
The access holes for fixing screws in prostheses on implants must be completed so that the screw is not damage if is necessary to remove the prostheses. Among the most common materials used are cotton, gutta percha and polytetrafuoroetylene tape. The aim of this study is to evaluate the biofilm formation in the materials described above, in order to establish a parameter that contributes to the choice of most suitable material. We used UCLAs, similar (4,1 mm) and titanium screws. The set was assembled with a torque of 32 N.cm. The materials were condensed inside the UCLAs and placed in culture medium with suspension of 3x 106 cells/ml of Candida albicans. The system was stored at 37 o C with shaking for 15 days and renewed every 48 hours. The quantification of biofilm was performed by MTT assay at 490 nm and reading. Analyzed the normal (p=0,304-Kolmogorov-Smirnov) and equal variances (p= 0,721- Scheffe). The ANOVA showed significant difference between groups (p<0,001) and Holm- Sidak significant difference was observed between the cotton and gutta groups (p<0,05) and cotton and PTFE (p<0,05), not significant difference between groups gutta and polytetrafuoroetylene tape (p>0,05), although values were higher polytetrafuoroetylene tape. Considering the limitations of this in vitro study, we conclude that:1. Both gutta-percha and polytetrafluorethylene tape showed less biofilm formation, with no statistically significant difference between the materials. 2. Cotton showed a level of biofilm formation significantly higher than the polytetrafluoroethylene tape and gutta percha. Therefore, further studies are necessary to confirm the limitations that this type of material can have when used as filling material of the screw access on the prosthetic implant.
Madiedo-Podvršan, Sabrina. "Development of a lung-liver in vitro coculture model for the risk assessment of inhaled xenobiotics." Electronic Thesis or Diss., Compiègne, 2022. http://www.theses.fr/2022COMP2703.
Urbanization and globalization are prevailing social phenomena that multiply and complexify the sources of modern pollution. Amongst others, air pollution has been recognized as an omnipresent life-threatening hazard, comprising a wide range of toxic airborne xenobiotics that expose man to acute and chronic threats. The defense mechanisms involved in hazardous exposure responses are complex and comprise local and systemic biological pathways. Due to this complexity, animal models are considered prime study models. However, in light of animal experimentation reduction (3Rs), we developed and investigated an alternative in vitro method to study systemic-like responses to inhalationlike exposures. In this context, a coculture platform was established to emulate interorgan crosstalks between the pulmonary barrier, which constitutes the route of entry of inhaled compounds, and the liver, which plays a major role in xenobiotic metabolism. Both compartments respectively comprised a Calu-3 insert and a HepG2/C3A biochip which were jointly cultured in a dynamically-stimulated environment for 72 hours. The present model was characterized using acetaminophen (APAP), a well-documented hepatotoxicant, to visibly assess the passage and circulation of a xenobiotic through the device. Two kinds of models were developed: (1) the developmental model allowed for the technical setup of the coculture, and (2) the physiological-like model better approximates a vivo environment. Based on viability, and functionality parameters the developmental model showed that the Calu-3 bronchial barrier and the HepG2/C3A biochip can successfully be maintained viable and function in a dynamic coculture setting for 3 days. In a stress-induced environment, present results reported that the coculture model emulated active and functional in vitro crosstalk that seemingly was responsive to high (1.5 and 3 mM) and low (12 and 24 μM) xenobiotic exposure doses. Lung/liver crosstalk induced modulation of stress response dynamics, delaying cytotoxicity, proving that APAP fate, biological behaviors and cellular stress responses were modulated in a broader systemic-like environment
Frederico, Éric Heleno Freire Ferreira. "Influência de um extrato aquoso de Coriandrum sativum na marcação in vitro de constituintes sanguíneos com tecnécio-99m e de sua associação com vibração gerada por plataforma oscilante na biodistribuição do radiofármaco Na99mTcO4 e na concentração de biomarcadores em ratos Wistar." Universidade do Estado do Rio de Janeiro, 2014. http://www.bdtd.uerj.br/tde_busca/arquivo.php?codArquivo=7550.
Coriandrum sativum, popularly known as coriander, is a vegetable used in human alimentation. It is also used as a medicinal plant for the treatment of diabetes, gastrointestinal complications, and as an antiedemic, antiseptic and emenagogue. In the investigations about the effects of a plant extract, it is important to determine some of its physicochemical parameters. Several experimental models have been used, including the use of radionuclides. In Nuclear Medicine procedures that assist the diagnosis of diseases, technetium-99m (99mTc) is the most used radionuclide. Red blood cells labeled with 99mTc are among the various cellular structures that can be labeled with this radionuclide and used as a radiopharmaceutical. The labeling process involving 99mTc requires the presence of a reducing agent and the most used for this purpose is the stannous chloride (SnCl2). The drugs therapy, diet conditions and diseases can alter the labeling of blood constituents, as well the biodistribution of several radiopharmaceuticals. The exposure to vibrations generated in oscillating platform produces whole body vibration (WBV) exercise. The aim of this study was to characterize the preparation of an extract of Coriandrum sativum, through physicochemical parameters, verify the effects of this natural product in radiolabeling of blood constituents and in association with vibration generated by platform on the Na99mTcO4 biodistribution, and in concentrations of some biomarkers. The extract of coriander had a pick absorbance at 480 nm. The coriander extract was inversely correlated with the concentration in electric conductivity. Was founded the highest value of pH at the lower concentration of the extract (0.5 mg/mL). There was no significant alteration on the labeling of blood constituents with 99mTc. The association between the coriander extract and vibration generated in platform with frequency of 12 Hz had effect in spleen, as observed in the fixation of the radiopharmaceutical in the organ and action in some organs altering the concentration of some biomarkers. In conclusion, physicochemical parameters can be useful to characterize the studied extract. Probably, the redox properties associated with the substances of this extract could be responsible by the absence of effect on the radiolabeling of blood constituents. The determination of the uptake of the radiopharmaceutical sodium pertechnetate in different organs permits to verify that extract of coriander alone was not capable in interfering on the biodistribution of the radiopharmaceutical. However, the treatment of the animals with vibration generated in the platform alters significantly the fixation of the sodium pertechnetate in the spleen and the concentrations of Cholesterol, triglyceride, CK and bilirubin.
Viray, Christina Marie. "Developing Innovative Bioengineering Platforms to Recapitulate Cell Microenvironments In Vitro." Thesis, The University of Sydney, 2021. https://hdl.handle.net/2123/24955.
Recha, Sancho Lourdes Georgina. "Development of biomaterial self-assembling based platforms to obtain human cartilage tissue in vitro." Doctoral thesis, Universitat Ramon Llull, 2016. http://hdl.handle.net/10803/394009.
El cartílago articular tiene una capacidad limitada de crecimiento y regeneración y, los tratamientos para restaurar la función del tejido, después de una lesión, son limitados y poco entendidos por la comunidad médica. Existe, por tanto, un gran interés en encontrar una solución práctica y agradable para el paciente que consiga la reparación del cartílago. La ingeniería de tejidos surgió para restaurar tejidos dañados usando nuevas plataformas terapéuticas basadas en células y/o biomateriales. Estas nuevas terapias pretenden crear estructuras similares al cartílago que imiten las propiedades mecánicas y biológicas que se dan in vivo. En este sentido, el uso de matrices biomiméticas que reproduzcan estructural y funcionalmente el microambiente nativo ha generado gran interés en este campo. Los péptidos auto-ensamblantes representan candidatos ideales para crear nichos celulares dado que, sus nanofibras y propiedades biomecánicas son similares a las de la matriz extracelular. En esta tesis, se han desarrollado nuevos biomateriales sintéticos con gran potencial para la reparación de cartílago. Éstos, están basados en el péptido auto-ensamblante RAD16-I decorado con motivos bioactivos, tratando de reproducir la matriz del cartílago. Dada la versatilidad del hidrogel RAD16-I, las nuevas matrices se formaron por simple mezcla del péptido RAD16-I con moléculas de heparina, condroitin sulfato y decorina. Estas matrices bi-compuestas presentan buena estabilidad química y estructural a pH fisiológico y son capaces de unir y liberar, gradualmente, factores de crecimiento. La evaluación de estas matrices se llevó a cabo mediante dos estrategias in vitro diferentes: la rediferenciación de condrocitos articulares humanos y, la inducción del linaje condrogénico en células madre derivadas de tejido adiposo. Ambos tipos celulares son considerados una buena fuente de células para obtener constructos que reparen defectos en el cartílago. Los resultados presentados en este trabajo muestran diferencias a nivel de comportamiento celular, patrones de expresión y propiedades mecánicas entre los dos tipos celulares y las diferentes condiciones de cultivo (matrices y medios). Cabe destacar que, ambos tipos celulares se diferencian a un linaje condrogénico en medio de inducción y que los constructos presentan propiedades mecánicas compatibles con un sistema condrogénico. Además, se ha determinado que la presencia de moléculas de heparina en la matriz promueve la supervivencia de las células madre derivadas de tejido adiposo. En conjunto, las nuevas matrices bi-compuestas representan un material fácil de preparar y prometedor para promover la diferenciación condrogénica. Por último, parte de esta tesis se ha centrado en el desarrollo de una nueva matriz compuesta mediante la infiltración del péptido RAD16-I con células en microfibras de policaprolactona (PCL). Se ha demostrado que esta nueva combinación ofrece una estructura funcional y biomimética, dado que, proporciona soporte mecánico por las fibras PCL y a su vez, facilita la adhesión y el crecimiento celular debido al hidrogel RAD16-I. El cultivo in vitro de condrocitos humanos desdiferenciados demuestra que la nueva matriz compuesta promueve la supervivencia celular y el restablecimiento del linaje condrogénico. En general, las propiedades sinérgicas de la nueva matriz compuesta proporcionan una plataforma terapéutica ideal para ayudar a la reparación del cartílago.
Adult articular cartilage has a limited capacity for growth and regeneration and, after injury, treatments to restore tissue function remain poorly understood by the medical community. Therefore, there is currently great interest in finding practical and patient-friendly strategies for cartilage repair. Tissue engineering has emerged to restore damaged tissue by using new cellular or biomaterial-based therapeutic platforms. These approaches aim to produce cartilage-like structures that reproduce the complex mechanical and biological properties found in vivo. To this end, the use of biomimetic scaffolds that recreate structurally and functionally the native cell microenvironment has become of increasing interest in the field. Self-assembling peptides are attractive candidates to create artificial cellular niches, because their nanoscale network and biomechanical properties are similar to those of the natural extracellular matrix (ECM). In the present thesis, new composite synthetic biomaterials were developed for cartilage tissue engineering (CTE). They were based on the non-instructive self-assembling peptide RAD16-I and decorated with bioactive motifs, aiming to emulate the native cartilage ECM. We employed a simple mixture of the self-assembling peptide RAD16-I with either heparin, chondroitin sulfate or decorin molecules, taking advantage of the versatility of RAD16-I. The bi-component scaffolds presented good structural and chemical stability at a physiological pH and the capacity to bind and gradually release growth factors. Then, these composite scaffolds were characterized using two different in vitro assessments: re-differentiation of human articular chondrocytes (ACs) and induction of human adipose derived stem cells (ADSCs) to a chondrogenic commitment. Both native chondrocytes and adult mesenchymal stem cells (MSCs), either bone marrow or adipose-tissue derived, are considered good cell sources for CTE applications. The results presented in this work revealed differences in cellular behavior, expression patterns and mechanical properties between cell types and culture conditions (scaffolds and media). Remarkably, both cell types underwent into chondrogenic commitment under inductive media conditions and 3D constructs presented mechanical properties compatible to a system undergoing chondrogenesis. Interestingly, as a consequence of the presence of heparin moieties in the scaffold cell survival of ADSCs was enhanced. Altogether, the new bi-component scaffolds represent a promising "easy to prepare" material for promoting chondrogenic differentiation. Finally, part of this thesis was focus on developing a composite scaffold by infiltrating a three-dimensional (3D) woven microfiber poly (ε-caprolactone) (PCL) scaffold with the RAD16-I self-assembling peptide and cells. This new combination resulted into a multi-scale functional and biomimetic tissue-engineered structure providing mechanical support by PCL scaffold and facilitating cell attachment and growth by RAD16-I hydrogel. The in vitro 3D culture of dedifferentiated human ACs evidenced that the new composite supports cell survival and promotes the reestablishment of the chondrogenic lineage commitment. Overall, the synergistic properties of the novel composite scaffold may provide an ideal therapeutic platform to assist cartilage repair.
Rohde, Rosemary Dyane Blake Geoffrey A. Heath James R. "Developing high-affinity protein capture agents and nanotechnology-based platforms for in vitro diagnostics /." Diss., Pasadena, Calif. : California Institute of Technology, 2009. http://resolver.caltech.edu/CaltechETD:etd-06032009-230415.
Cimetta, Elisa. "Design and development of microscale technologies and microfluidic platforms for the in vitro culture of stem cell." Doctoral thesis, Università degli studi di Padova, 2009. http://hdl.handle.net/11577/3426444.
Le impellenti necessità legate allo sviluppo di nuovi farmaci e di terapie innovative per la cura di malattie dell’apparato muscolare, quali ad esempio la Distrofia Muscolare di Duchenne o l’infarto miocardico, hanno portato alla crescente domanda di nuovi metodi e tecnologie. E’ chiaro inoltre come le cellule staminali possano costituire una risorsa fondamentale per la generazione di tessuti umani artificiali da impiegare in tali processi. Tale prospettiva, richiederebbe da un lato strumenti sofisticati per il controllo e il differenziamento delle cellule staminali e dell’altro, l’integrazione di questi all’interno di procedure in grado di soddisfare i requisiti fondamentali dei modelli sui quali operare per lo sviluppo di nuovi farmaci o strategie terapeutiche. Tra i requisiti fondamentali da rispettare si citano quindi: micronizzazione, versatilità, basso costo e highthroughput. Obiettivo fondamentale di questa tesi sono stati la progettazione, sviluppo e fabbricazione di tecnologie su scala micrometrica in grado di riprodurre una stimolazione biomimetica ispirata al microambiente cellulare in vivo e, nello stesso tempo, di rispondere ai requisiti tecnologici descritti sopra. E’ stata effettuata un’analisi semi quantitativa basata sull’analisi dei tempi caratteristi dei fenomeni su microscala, che ha portato alla produzione di diagrammi operativi da impiegarsi nelle fasi di progettazione e sviluppo di tali strategie sperimentali. Sono stati sviluppati microbioreattori all’interno di piattaforme microfluidiche applicate poi allo studio di sistemi cellulari. In particolare, sono stati effettuati studi biologi sull’effetto di gradienti di concentrazione all’interno dell’importante signaling cellulare del Wnt. E’ stata realizzata una tecnica per l’organizzazione topologica su microscala di colture cellulari su substrati in hydrogel dalle proprietà meccaniche definite. Le tecnologie sviluppate sono state impiegate per casi studio dall’elevato valore scientifico e sono state interfacciate con colture di particolare interesse quali mioblasti umani distrofici e cellule cardiache derivate da staminali embrionali umane. Infine, è stato proposto un prototipo di piattaforma microfluidica in grado di accoppiare le stimolazioni di tipo topologico al controllo dell’ambiente solubile su colture cellulari. I risultati ottenuti aprono nuove ed interessanti prospettive sia per lo sviluppo di nuovi farmaci che di strategie terapeutiche volti alla cura di patologie ed allo studio approfondito della complessità dei sistemi biologici.
Fatanat, Didar Tohid. "Lab-on-Chip platforms: enabling technology for label-free detection, separation, patterning and «in vitro» culture of cells." Thesis, McGill University, 2013. http://digitool.Library.McGill.CA:80/R/?func=dbin-jump-full&object_id=117116.
Les dispositifs biomédicaux développés pour la détection, le triage et la culture cellulaire sont des outils importants en diagnostic clinique et en recherche fondamentale. Récemment, avec les progrès dans le domaine de la miniaturisation et les microfluidiques, les 'laboratoires-sur-puces' (LOC) ont commencé à jouer un rôle significatif dans la détection et l'enrichissement des différents types de cellules. Parmi les multitudes LOC développés pour ces applications, ceux n'impliquant pas de marquage représentent une option attrayante, car ils ne modifient pas les propriétés des cellules ciblées par le triage. Séparation à base d'adhérence cellulaire ou en utilisant la différence entre la taille physique des cellules sont les deux approches principales qui ne nécessitent pas de prétraitement des cellules et leur marquage. Dans cette thèse, des micropuces proposées emploient des méthodes sans marquage préalable des cellules ciblées tout en exploitent, soit leurs propriétés d'adhésion et leur affinité pour la biointerface, soit leur différence de taille par rapport la taille des autres cellules dans le mélange initial. En ce qui concerne la première micropuce, nous avons développé un dispositif microfluidique original et multicouche à base de thermoplastique pour la séparation de micro/nanoparticules et de cellules de tailles différentes. Une efficacité de séparation supérieure à 95% a pu être réalisée à haut débit (150 μl/min),Quant à la séparation basée sur l'adhérence, tout d'abord, nous avons introduit des LOCs pour produire des gradients de concentration de biomolécules dans un seul canal microfluidique. Dans un deuxième temps, ce concept a été utilisé pour fabriquer une puce multifonctionnelle sur laquelle, il était possible de simultanément capturer, séparer des cellules rares de source primaire, paver la surface avec des motifs des cellules et les cultiver sur la même puce. En utilisant cette puce, des cellules progénitrices d'oligodendrocytes (OPC) et des cardiomyocytes provenant respectivement du cerveau et du cœur du rat, ont pu être séparés en 10 min avec une efficacité de séparation supérieure à 95% des autres cellules dans le mélange tissulaire. La séparation de ces deux types de cellules primaires démontre l'efficacité et l'universalité de cette puce multifonctionnelle pour la séparation d'une gamme de mélanges cellulaires avec différentes concentrations initiales des cellules ciblées par le triage.
Pliquett, Jacques. "Development of fluorescent platforms for the design of multifunctional compounds for in vitro and in vivo applications in molecular imaging." Thesis, Bourgogne Franche-Comté, 2018. http://www.theses.fr/2018UBFCK067.
The objective of this thesis was the development and evaluation of new molecular platformsfor optical fluorescence imaging applications. This work sought to develop new tools that caneasily be modified and adapted to the specific needs of the intended use. This is required asthe fluorophore will influence the final properties and should thus be incorporated beforestructural optimization of the selected agent rather than at the very end. Two main axes wereexplored; the use of BODIPYs for the development of trackable therapeutic agents that areprimarily intended for in vitro applications and the use of azaBODIPYs for the design of an invivo compatible fluorescent platform.In the first part two fluorophores on the basis of a 3,5-dichloro-BODIPY were identified aspromising platforms. These platform molecules were selectively functionalized using a gold(I)-phosphine moiety, a thiosugar and a phosphonium to explore their selective functionalizationand investigate the influence of each substitutents position on the final properties. Weshowed that a site-specific, selective functionalization with these fragile substituents ispossible and developed 12 gold(I)-bearing therapeutic agents. We evaluated thephotophysical properties of all obtained compounds which was followed by a characterizationof their biological properties (antiproliferative properties on 3 cancer cell lines, lipophilicbalance and cellular gold accumulation as well as fluorescence imaging on 3 cell lines for upto 24h). We succeeded in developing a panel of closely related trackable compounds thatdisplay mixed activity in cells and distinct cellular localization. This investigation permitted theselection of three to four hits that will be studied further.In the second part we developed an in vivo-compatible multifunctional platform following twostrategies: the first was the use of 1,7-di(phenol)-3,5-di(phenyl)-azaBODIPY and thefunctionalization of the hydroxy groups for the development of a bioconjugable NIR-I probe.Unfortunately the developed probe displayed very unfavourable optical properties; wetherefore developed a new strategy that is entirely based on the functionalization of the boronatom. Using this approach we successfully synthesized 2 watersoluble, strongly fluorescent(NIR-I) molecular platforms that were conjugated to an innovative antibody to image the PD-L1 ligand. The developed probes displayed excellent optical properties, are stable for at least48h in mice plasma and were validated in a preclinical study on mice. The developed probesdisplayed strong fluorescence in vivo and showed no acute toxicity.The developed methodology shows great potential for further investigations and futurestudies; it can be transposed onto other closely related fluorophores and permits versatilefunctionalization with a large variety of compounds of interest. Its use is thus not limited tobiological, biochemical and medical applications
Griffiths, Natalie Helen. "Mechanisms of GABAA and glycine receptor analgesia in the spinal dorsal horn : in vitro models as translational platforms for drug discovery." Thesis, University of Leeds, 2015. http://etheses.whiterose.ac.uk/12560/.
Srinivasan, Rajesh. "Platform Technologies for In vitro Point-of- Care Diagnostics." Thesis, 2022. https://etd.iisc.ac.in/handle/2005/5737.
Lai, Tzu-Hsiang, and 賴子詳. "An antibiotic delivery platform using in vitro packed AP205 VLPs." Thesis, 2019. http://ndltd.ncl.edu.tw/handle/9ww673.
國立臺灣大學
生化科學研究所
107
Nowadays, small molecular drugs are used for chemotherapy to treat cancer diseases despite that these small molecules are non-targeting drugs. However, all efficacious drugs have adverse side effects. In order to avoid these unwanted hazards, scientists dedicate efforts to develop a carrier, which is capable of delivering drugs to target cancer cells precisely instead of anonymously. VLPs (Virus like particles) is a nanoscale self-assembled protein-based cage that is composed of only capsid from virus. The surface of capsid is tolerated with chemical modifications or genetic engineering to present the targeting peptides and epitopes. By far, the Qβ, MS2, and HBVc VLPs have been extensively studied in application of vaccine and drug delivery. In this study, the AP205 VLPs are used to encapsulate two small molecular drugs, doxorubicin (dox) and neomycin (neo), through the RNA hairpin-capsomere interactions. The encapsulated RNA hairpin is phosphorothioated by T4 PNK reaction, and then covalently coupled with dox or neo through amine-to-sulfhydryl crosslinker. The AP205 VLPs dox and AP205 VLPs neo are packed successfully in vitro. However, both the in vitro packed VLPs show low cytotoxicity towards cancer cells, which indicates that low coupling efficiency of dox/neo to the RNA hairpin, contributing to a low level of compound loading within the VLPs. The dox/neo and RNA hairpin coupling require further investigations in future works.
Lin, Meng Bo, and 林孟柏. "Design and Development of An In-Vitro Ultrasonic Stimulation Testing Platform." Thesis, 2016. http://ndltd.ncl.edu.tw/handle/20041466419041108065.
Pereira, Ana Rita Oliveira Alves. "Modelling breast cancer metastatic bone niche: a novel in vitro 3D microfluidic platform." Master's thesis, 2016. https://hdl.handle.net/10216/90164.
Mei, Hsiang-I., and 梅湘怡. "In-vitro Er:YAG laser treatment platform construction with the implant for peri-implantitis." Thesis, 2017. http://ndltd.ncl.edu.tw/handle/6w6p59.
國立陽明大學
生物醫學工程學系
105
Peri-implantitis, which is induced by dental biofilm leading to the periodontal immune reaction, subsequently causes peri-implantitis inflammation and bone loss. The peri-implantitis is usually treated by using conservative mechanical debridement, air abrasives, antiseptic treatment, laser etc. The Er: YAG laser is a 2940nm wavelength laser with the highest energy insertion by closest connection to water among the existing lasers. This dental laser has high potentials for peri-implantitis treatment. Unfortunately, there has been no in-vitro laser experiment platform that considers different implant positions and the levels of peri-implantitis diseases using a real dental implant with a threaded surface. The standard clinical adult tooth jaw model was scanned to construct the digital model with 2mm and 6mm bone loss depth mimicking serious peri-implantitis at the incisor, first premolar, and first molar. This study constructs a standard in-vitro laser treatment platform with dental implant surface on which bacterial adhesion for peri-implantitis at different tooth positions. An Er: YAG laser, working with a chisel type glass tip, was moved from the buccal across the implant thread to the lingual for about 60 seconds per sample to verify the in-vitro laser treatment platform. The result showed that the sterilization rate can reach up to 90% and the jaw model was not damaged after laser irradiation testing.This study concluded that using integrated image processing, reverse engineering, CAD system and a 3D printer to construct a peri-implantitis model replacing the implant on bacterial adhesion and acceptable sterilization rate proved the feasibility of the proposed laser treatment platform.
Pereira, Ana Rita Oliveira Alves. "Modelling breast cancer metastatic bone niche: a novel in vitro 3D microfluidic platform." Dissertação, 2016. https://hdl.handle.net/10216/90164.
Scamardella, Sara. "A NEW HUMAN SKIN EQUIVALENT MODEL AS IN VITRO TESTING PLATFORM FOR BIOACTIVE MOLECULES." Tesi di dottorato, 2015. http://www.fedoa.unina.it/10401/1/Scamardella_Sara.pdf.
Langella, Angela. "Fabrication of in vitro epithelial tissues as a testing platform for drug delivery systems." Tesi di dottorato, 2017. http://www.fedoa.unina.it/12097/1/Langella_Angela_phd_thesis.pdf.
Addae-Mensah, Kweku A. "A microfabricated microcantilever array a platform for investigation of cellular biomechanics and microforces in vitro /." Diss., 2008. http://etd.library.vanderbilt.edu/available/etd-08182008-153315/.
Chien, Chi-Chen, and 簡基城. "Studies of Immunomodulation Effect of Ganoderma lucidum Using an in vitro Cord Blood MNC-Based Platform Method." Thesis, 2004. http://ndltd.ncl.edu.tw/handle/6kq78m.
國立清華大學
生命科學系
92
An in vitro model for testing natural substances is developed in this study. The model used mononuclear cells (MNCs) from human umbilical cord blood (hUCB) for phenotypic expression induction platform. Natural substances, such as Ganoderma or grape seed proanthocyanidins extract, were added to the medium and then cultured for seven days. The stem cells/progenitors contained in the mononuclear cells were modulated by the active ingredients and changed in the course of differentiation and maturation. Flow cytometry was used for phenotypic analysis. Meanwhile, we utilized a cell-fluid based microchip on Agilent Bioanalyzer, and discovered an alternative assay method for phenotypic expression studies. Such a measurement could analyze the phenotypes of cells in a small population as few as 200 cells with good sensitivity and accuracy. We found that Ganoderma lucidum extract could enhance the NK cells composition in immune cell subpopulation when used to treat hUCB MNCs. After treatment, the enriched NK cells preserved similar cytotoxicity function. Enrichment of specific subsets of immune cells enabled us to explore further studies, such as investigating the optimal effector-to-target cells ratio in this study, or potential future systems biology experiments during differentiation courses. The model was also utilized to screen different sort of natural substances. We found that, for example, wheat grass extract up-regulated CD 56+ NK cell composition, whereas rutin and quercetin down-regulated instead. Different fractions of natural substances could be further testified quickly with this platform. Furthermore, we observed no significant change as the concentration of natural substance was below a critical concentration. The optimal oral consumption dosage of a specific natural health product could be predicted. Taken together, we demonstrated that this technique provides a good platform to study natural substances. The responses of stem cells/progenitors to natural substances, the alteration in phenotypic expression of immune cell subpopulation, and the modulation of human immunity could be further explicated using this technique.
Magalhães, Marta Isabel Ricardo de. "Development of an in vitro platform for a 3D microphysiological systems of human iPS-derived endothelial cells." Master's thesis, 2021. http://hdl.handle.net/10362/120566.
Neste trabalho apresentamos o desenvolvimento original de um chip para co-cultura de células para a construção de redes microvasculares 3D. O layout do chip foi projetado no Autodesk AutoCAD2020 para obter 3 câmaras de tecido interconectadas onde as células serão semeadas e irão crescer. Os compartimentos das células são interligados e alimentados com o meio de cultura através dos poros laterais em forma de ampulheta (dois poros para cada câmara). As câmaras celulares estão conectadas à entrada/saída de meio, que é entregue por um sistema de alimentação automático. Um dos objetivos iniciais deste trabalho era fazer com que os vasos crescessem em direção a ambos os poros, para isso um gradiente de pressão era necessário. Um sistema de entrega de meio foi projetado e construído com uma impressora 3D e operado por um atuador digital com uma placa microcontrolador. O sistema oscilante mantém uma diferença de nível (pressão estática) entre os dois recipientes do meio de cultura, mudando as suas posições automaticamente por intervalos de tempo previamente definidos, promovendo um crescimento celular uniforme. A diferença de altura do meio de cultura e o tempo entre as mudanças podem ser ajustados pelo sistema de controle programável. Para além disso, o design do layout do chip permite o crescimento de células para simular uma rede 3D de vasos sanguíneos. Após a semeação e polimerização do gel de fibrina, as células cresceram ocupando todo o espaço da câmara. Isto pode ser observado por microscopia sem interrupção da cultura, pois o suporte do chip é transparente. Estes indicam que, uma plataforma adequada para ensaios biológicos usando diferentes tipos de células em co-cultura (por exemplo, fibroblasto com células endoteliais humanas) pode ser realizada.
Gioiella, Filomena. "A new bioengineered 3D tumor platform in vitro to replicate tumor-stroma interaction and investigate anti-cancer drug delivery." Tesi di dottorato, 2016. http://www.fedoa.unina.it/10856/1/Gioiella_Filomena_28.pdf.
Ramappa, Nirmala Kuppalu. "A system biology approach to study pancreatic ductal adenocarcinoma (PDAC) cells in in vitro culture." Doctoral thesis, 2018. http://hdl.handle.net/2158/1116922.