La première partie de cette thèse est dédiée à la modification de polyélectrolytes pour former des films de multicouche de polyélectrolytes (PEM) ayant des propriétés d’adhésion de protéine et de cellules bien contrôlées et modifiables par étirement. L’acide polyacrylique a été modifié avec des groupes latéraux phosphorylcholine (PC) à des taux de 25 % (PAA-PC) ou avec des chaînes oligo(éthylène oxyde) terminées par la biotine : (EO)nBiotine (n = 0, 3, 9 et 18) avec de taux de modification de 1, 5, 10 ou 25 %. Des PEM incorporant ces polymères lient spécifiquement la streptavidine et repoussent tout autre protéine. Les propriétés d’adsorption et la sélectivité de ces PEM ont été mesurées par microbalance à quartz. Sur un substrat de PDMS étirables, on a construit des PEM terminés par un PAA portant des RGD recouvert par deux couches contenant PAA-PC. Au repos, seuls les PC sont exposés et inhibent l’adhésion cellulaire ; sous étirement, les groupes RGD sous-jacents sont exposés et déclenchent l’adhésion de fibroblastes.La deuxième partie est consacrée à l‘étude d’acide polyméthacrylique modifié hydrophobiquement avec des chaînes alkyle liées par des esters à la chaîne principale. 3 chaînes différentes ont été greffées : -C12H25 ; -C18H35 et C4H8-OOC- C11H23 avec des taux de 1, 5 and 10 %. Ces polymères sont associatifs et forment des hydrogels dans des tampons physiologiques pour des taux de modifications de 5% et des concentrations supérieures à 4% en poids. Ces gels ont été caractérisés par des mesures rhéologiques. Leur incubation avec des lipases provoque une baisse de leur viscosité, interprétable par une coupure des esters. Quand les gels faits à partir du PAA-C12 sont incubés avec une culture de Pseudomonas aeruginosa, la viscosité baisse également, ce qui montre que les chaînes sont également coupées in vivo. / The first part of this thesis is dedicated to the modification of polyelectrolytes to form polyelectrolyte films with controlled and stretch responsive cell and protein adsorption properties. Poly(acrylic acid) (PAA) was modified with side phosphorylcholine groups (PC) at rates of 25 % or with oligo(ethylene oxide) chains ended by biotin ((EO)nBiotin, (n =0, 3, 9 and 18) at 1, 5, 10 and 25 % modification rates. Polyelectrolytes multilayer films (PEM) containing these polyelectrolytes bind selectively streptavidin but repel all other proteins. The adsorption properties and selectivity were measured by quartz crystal microbalance. On a stretchable PDMS substrate, we have built PEM ended by PAA bearing RGD, covered by two PAA-PC layers on the top. Under rest, only the PC groups are exposed and prevent cell adhesion; when the film is stretched, the underlying RGD groups are exposed, and trigger adhesion of fibroblasts.The second part was consecrated to the study of poly(methacrylic acid) hydrophobically modified with alkyl chains connected through an ester moiety to the main chain. Three different chains were grafted -C12H25; -C18H35 and -C4H8- OOC-C11H23 with a rate of 1, 5 and 10 %. These polymers associate in water and form hydrogels in physiological buffer, for modification rates higher than 5 % and polymer concentrations higher than 4 wt. %. The gels were characterized by rheology. Their incubation with lipases resulted in a decrease of their viscosity, which could be interpreted by the cleavage of the hydrophobic side chains, by rheological tests. When the gels with PAA-C12 were incubated with a culture of Pseudomonas aeruginosa, their viscosity decreased, which shows that alkyle chains are also cleaved in vivo.

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Limb-girdle muscular dystrophy type 2B (LGMD2B) is caused by mutations in the dysferlin gene, resulting in non-functional dysferlin, a key protein found in muscle membrane. Treatment options available for patients are chiefly palliative in nature and focus on maintaining ambulation. Our hypothesis is that galectin-1 (Gal-1), a soluble carbohydrate binding protein, increases membrane repair capacity, myogenic potential, M2 macrophage polarization and decreases NF-κB inflammation in dysferlin-deficient models. To test this hypothesis, we used recombinant human galectin-1 (rHsGal-1) to treat dysferlin-deficient models. We show that rHsGal-1 treatments of 48 h-72 h promotes myogenic maturation as indicated through improvements in size, myotube alignment, and myoblast migration in dysferlin-deficient myotubes. Furthermore, rHsGal-1 showed an increased membrane repair capacity of dysferlin-deficient myotubes. Improvements in membrane repair after only a 10 min rHsGal-1treatment suggests mechanical stabilization of the membrane due to interaction with glycosylated membrane bound, ECM or yet to be identified ligands through the CDR domain of Gal-1. rHsGal-l significantly reduces canonical NF-κB inflammation through TAK 1, P65, P50. Lastly we find 2.7 mg/kg in vivo rHsGal-1 treatment in BLA/J mice supports an M2 cyto-regenerative macrophage populations. Together our novel results reveal Gal-1 remediates disease pathologies in LGMD2B through changes in integral myogenic protein expression, mechanical membrane stabilization, immune modulation, and reducing canonical NF-κB inflammation.

Optical microscopy is the simplest and the gold standard method adopted for the screening and subsequent diagnosis of various hematological and infectious diseases like malaria, sickle cell disease, tuberculosis etc. In addition to infectious disease diagnosis, its applications range from routine blood tests to the more sophisticated cancer biopsy sample analysis. Microscopy Tests (MTs) follow a common procedural workflow: (1) A technician prepares a smear of the given sample on a glass slide in a specific manner depending on the sample and the disease to be diagnosed; (2) The smeared slide is subsequently exposed to fixative agents and different histochemical stains specific to the diagnosis to be performed and (3) the prepared slide is then observed under a high quality bright- field bench-top microscope. An expert pathologist/cytologist is required to manually examine multiple fields-of-views of the prepared slide under appropriate magnification. Multiple re-adjustments in the focus and magnification makes the process of microscopic examination time consuming and tedious. Further, the manual intervention required in all the aforementioned steps involved in a typical MT, makes it inaccessible to rural/resource limited conditions and restricts the diagnostics to be performed by trained personnel in laboratory settings. To overcome these limitations, there has been considerable research interest in developing cost-effective systems that help in automating MTs. The work done in this thesis addresses these issues and proposes a two-step solution to the problem of affordable automation of MTs for cellular imaging and subsequent diagnostic assessment. The first step deals with the development of a low cost portable system that employs custom-built microscopy setup using o -the-shelf optical components, low cost motorized stage and camera modules to facilitate slide scanning and digital image acquisition. It incorporates a novel computational approach to generate good quality in-focus images, without the need for employing high-end precision translational stages, thereby reducing the overall system cost. The process of slide analysis for result generation is further automated by using image analysis and classification algorithms. The application of the developed platform in automating slide based quantitative detection of malaria is reported in this thesis. The second aspect of the thesis addresses the automation of slide preparation. A major factor that could influence the analysis results is the quality of the prepared smears. The feasibility of automating and standardizing the process of slide preparation using Microfluidics with appropriate surface fictionalization is explored and is demonstrated in the context of automated semen analysis. As an alternative to the mechanism of fixing the spermatozoa to the glass slide by smearing and chemical treatment with fixative, microfluidic chips pre-coated with adhesive protein are employed to capture and immobilize the cells. The subsequent histochemical staining is achieved by pumping the stains through the microfluidic device. The proof-of-principle experiments performed in this thesis demonstrate the feasibility of the developed system to provide an end-to-end cost-effective alternative solution to conventional MTs. This can further serve as an assistive tool for the pathologist or in some cases completely eliminate the manual intervention required in MTs enabling repeatability and reliability in diagnosis for clinical decision making