Spelling suggestions: "subject:"apolymer microarray"" "subject:"expolymer microarray""
1 |
Biomedical applications of polymer microarraysVenkateswaran, Seshasailam January 2017 (has links)
In my PhD polymer microarrays have been central in discovery of new materials for cardiovascular repair, cartilage tissue engineering and bacteria resistant medical devices. This has led to the work described in the following four chapters of my thesis. In the first part of my thesis polymers for the development of novel heart valve leaflets were identified. Diseased heart valves are currently replaced with the either synthetic or bioprosthetic (acellular xenografts) valve prostheses. While synthetic prosthesis have excellent durability, thromboembolic complications are frequent, requiring patients to undergo lifelong anti-coagulation therapy. On the other hand, the leaflets of bioprosthetic valves undergo structural deterioration, resulting in the patients having to undergo follow-up replacement surgeries. In order to overcome these shortcomings, the aim of this part of my PhD was to discover polymers that will enable the development of a ‘bio-synthetic’ heart valve, with the durability of synthetic valves and the biocompatibility of bioprosthetic vales. Polymers that bind valve interstitial cells (cells with a plastic fibroblast / myofibroblast phenotype that renew the extracellular matrix components of the valve leaflets) and also enable stable expression of key markers were identified. Immunohistochemistry and RNA expression analysis identified polymers for coating 3-D scaffolds, with the coated scaffolds showing excellent cell invasion, viability and maintenance of valve interstitial cell markers. To mimic the regions of the valve leaflets with differing stiffness, the response of valve interstitial cells to substrate stiffness was studied with various crosslinked gels. Thus, polymeric gels, prepared with the same chemical composition but with different Young’s modulus (covering 3 orders of magnitude) showed valve interstitial cell attachment with the cells showing differing behaviour based on the stiffness of the gels. In the second part of this thesis, polymers were identified for cartilage repair. Hyaline articular cartilage has very low potential for self-renewal, therefore cell-based therapies with autologous chondrocyte implantation are desired. Due to limited availability from biopsies, chondrocytes have to be expanded by in vitro culture; and fully defined synthetic culture substrates are essential for regulatory approvals. Using the high throughput approach I identified ‘hit’ polymers that allowed adhesion, proliferation and long-term culture of primary human chondrocytes and also chondrocytes derived from Mesenchymal stem cells. 2-D scale-up identified 2 lead polymers that supported long-term attachment and maintenance of chondrocyte markers. Since prolonged monolayer culture is known to induce loss of chondrocyte phenotype (dedifferentiation), 3D versions of the polymers were prepared and their potential for their long-term maintenance of chondrocytes via immunohistochemistry and RNA expression was demonstrated. The 3D gels were also used to encapsulate chondrocytes and their long-term maintenance of phenotype within these matrices, offers the exciting possibility of using these matrices for cartilage regeneration. The third part and fourth parts of the thesis focussed on reducing medical device associated infections. Thus polymers identified that prevented binding of a variety of bacteria including clinical isolates from infected medical devices, were used to coat two commercially available central venous catheters resulting in up to 96% reduction in bacterial binding. This non-binding was enhanced by the generation of polymeric nanocapsules containing the anti-bacterial eugenol (or its natural source clove oil). A coating consisting of eugenol nanocapsules entrapped within an interpenetrating network of the best bacteria repellent polymer, allowed slow-release of eugenol and further improved its performance.
|
2 |
Polymer microarrays for cell based applicationsHansen, Anne Klara Brigitte January 2012 (has links)
The development and identification of new biomaterials that can replace specific tissues and organs is desirable. In the presented PhD thesis polymer microarrays were applied for the screening of polyacrylates and polyurethanes and evaluation for material discovery for applications in the life sciences. In the first part of the thesis, the largest polymer microarray ever made with more than 7000 features was fabricated and subsequently used for the screening of polyacrylates that can control the fate of human embryonic stem cells. As stem cells have unique properties that offer the potential of replacing damaged or diseased tissue in future, the identification of cultivation substrates that can replace current biological and animal derived products was desirable. The water contact angle, roughness and cell doubling time of the cells on the identified polymers was determined and the stem cells characterised after 5 passages and compared to the currently most widely used animal derived substrate MatrigelTM. In the second part of the thesis, the development of a new polymer gradient microarray is presented. Initial studies involved the optimisation of printing parameters for the generation of linear polymer gradient lines and confirmed by XPS analysis. Cellular binding studies with the suspension cell line K562 and the adherent cell line HeLa were carried out and compared to previous binding studies to confirm the success of the concept. In further studies, the polymer gradients were functionalised with small molecules and proteins, allowing the generation of a protein gradient microarray with Semaphorin 3F. In binding studies with neuron cells it could be shown that the binding of the cells was concentration-dependent. The identification of polyacrylates for the effective and rapid activation and aggregation of platelets is described in the third part of the presented thesis. Here, polymer microarrays were applied for the binding of platelets in human blood samples. The amount of bound platelets as well as their activation state was compared to the natural agonist collagen by employing fluorescence intensity studies and scanning electron microscopy. In shear studies, the activation of the platelets by the polymers was evaluated under physiological conditions. The mechanism by which the polymer triggered the activation was further explored by protein binding studies. It was shown that the initial adsorption of fibrinogen and von Willebrand factor on the polymers lead to the adherence and aggregation of platelets. In the final part of the presented thesis, polymer microarrays were used to identify polymers that can sort and collect the precursor cells of platelets (megakaryocytes). For this purpose, the cell lines K562 and MEG-01 were used as cellular models. The identified polymers and the effect on the immobilised cells was further investigated by scanning electron microscopy, flow cytometry and miRNA studies. The adsorbed proteins on the different polymers were found to influence the cellular morphology on the different substrates.
|
3 |
Polymer microarrays for biomedical applicationsSimmonte Owens, Matthew John January 2017 (has links)
Biocompatible polymers are used exhaustively within the biomedical arena, demonstrating a mechanical and chemical diversity that few other materials possess. As polymer technologies evolves to cater for new medical demands, even the most niche biomedical application becomes an achievable reality. However, the discovery of new polymers is hindered by the complexity and intricacy in which the biological milieu interacts with a new substrate, reducing the ability to predict the appropriateness of a certain polymer for a specific application. This drawback can be countered by the high-throughput evaluation of large numbers of chemically diverse polymer candidates. In this thesis, the use of polymer microarrays is invoked to address two separate medically-relevant issues: the control of inflammation, and the improvement of cancer screening. In addition, I provide details of how polymer microarray techniques and technology can be employed to expand the repertoire of biomaterials research. Mitochondrial DNA (mtDNA) is an alarm molecule that contributes to the cytokine storm observed during severe tissue injury. An application where control of this systemic inflammation is achieved through scavenging of mtDNA by a polymer was proposed. Primary screening highlighted that 166 out of the 380 polymers evaluated bound to blood cells, making them unsuitable for a blood-based application. The remaining 214 blood-compatible polymers were cross-examined for mtDNA binding. Through polymer microarray and subsequent scale-up of promising candidates, a poly(methoxyethyl methacrylate-co-di(ethylamino)ethyl acrylate-co-methoxyethyl acrylate) was found to have a remarkable ability to scavenge mtDNA. Removal of cell-free mtDNA using this polymer is proposed to remove a key trigger of systemic inflammation. Cervical cancer screening includes the cytological evaluation of patient material for developed or developing abnormalities. An application was sought that would enrich for cancerous/pre-cancerous cells and improve upon current standards for detection. Four cancerous cervical cell lines (HeLa, CaSki, SiHa, and C33a) and four precancerous cell lines (W12E, W12G, W12GPX, and W12GPXY) were interrogated to identify polymers with consistent binding that may improve routine cytological evaluation. A short-list of 24 polymers was assembled, and cells from liquid based cytology samples from healthy patient were spiked with DiI-labelled cancerous/precancerous cells and the short-listed polymers were re-evaluated for preferential binding. An enrichment of abnormal cervical cells was observed with three polymers, which could form the foundation for improved screening resources. Inkjet printing can be a useful tool in developing patterned substrates, such as polymer microarrays. A piezoelectric drop-on-demand printer was used to explore the methods in which these can be fabricated. A wettability assay using picolitre volumes was developed and used to characterise O2 plasma treatment of glass slides. Additionally, the printing of a cell-binding polymer using this approach enabled the decoration of cells with precise spatial resolution.
|
4 |
Biocompatible polymer microarrays for cellular high-content screeningPernagallo, Salvatore January 2010 (has links)
The global aim of this thesis was to study the use of microarray technology for the screening and identification of biocompatible polymers, to understand physiological phenomena, and the design of biomaterials, implant surfaces and tissue-engineering scaffolds. This work was based upon the polymer microarray platform developed by the Bradley group. Polymer microarrays were successfully applied to find the best polymer supports for: (i) mouse fibroblast cells and used to evaluate cell biocompatibility and cell morphology. Fourteen polyurethanes demonstrated significant cellular adhesion. (ii) Analysis of the adhesion of human erythroleukaemic K562 suspension cells onto biomaterials with particular families of polyurethanes and polyacrylates identified. A DNA microarray study (to access the global gene expression profiles upon cellular binding) demonstrated that interactions between cells and some polyacrylates induced a number of transcriptomic changes. These results suggested that, during these interactions, a chain of cellular changes is triggered, most notably resulting in the downregulation of membrane receptors and ligands. (iii) Identification of polymers with potential applications in the field of stem cell biology. Polymers were identified that showed attachment, promotion and stabilisation of hepatocyte-like cells. A polyurethane support (PU-134) was pinpointed, which significantly improved both hepatocyte-like cell function and “lifespan”. A second project investigated biomaterials that promoted adhesion, growth and function of endothelial progenitor cells. A new polymer matrix was identified which contained the necessary signals to promote endothelial phenotype and function. This has potential application in the creation of blood vessels and the endothelialisation of artificial vessel prostheses and stent coatings for improving angioplasty therapy. (iv) The study of bacterial adhesion, focusing on the adhesion of food-borne pathogenic bacterium Salmonella enterica serovar typhimurium, strain SL1344, and the commensal bacterium Escherichia coli, strain W3110. Several polymers were found to support selective bacterial enrichment, as well as others that minimised bacterial adhesion.
|
5 |
Bacterial attachment to polymeric materials correlates with molecular flexibility and hydrophilicitySanni, O., Chang, Chien-Yi, Anderson, D.G., Langer, R., Davies, M.C., Williams, P.M., Williams, P., Alexander, M.R., Hook, A.L. 09 December 2014 (has links)
Yes / A new class of material resistant to bacterial attachment has been discovered that is formed from polyacrylates with hydrocarbon pendant groups. In this study, the relationship between the nature of the hydrocarbon moiety and resistance to bacteria is explored, comparing cyclic, aromatic, and linear chemical groups. A correlation is shown between bacterial attachment and a parameter derived from the partition coefficient and the number of rotatable bonds of the materials' pendant groups. This correlation is applicable to 86% of the hydrocarbon pendant moieties surveyed, quantitatively supporting the previous qualitative observation that bacteria are repelled from poly(meth)acrylates containing a hydrophilic ester group when the pendant group is both rigid and hydrophobic. This insight will help inform and predict the further development of polymers resistant to bacterial attachment. / Wellcome Trust (grant number 085245) and EMRP (IND56)
|
Page generated in 0.0429 seconds