Collagen type I is an extracellular matrix (ECM) protein that affords tensile strength and biological scaffolding to numerous vertebrate and invertebrate tissues. This strength has been attributed to the triple-helical structure of the collagen type I molecules, their organization into fibrils, and the presence of inter-molecular, covalent, enzymatic crosslinks. There are several different types of these crosslinks; their composition is tissue-specific and dependent upon factors such as age and health. Furthermore, these enzymatic crosslinks tend to form specifically at amino/N- and carboxy/C-terminal crosslinking sites. The mechanical behavior of collagen type I has been investigated, via experiment and theory, at the level of the molecule, microfibril, fibril, and fiber. However, the influence of different enzymatic crosslinks and their location (e.g., N- vs. C-site) on the mechanics of collagen type I has not been investigated in the literature. We employed molecular dynamics to model the mechanical behavior of uncrosslinked and crosslinked ~23-nm-long molecular segments and ~65-nm-long microfibril units of collagen type I. We then used these molecular simulations to construct a model of a single collagen type I fibril by considering the ~65-nm-long microfibril units arranged in series and then in parallel. When a uniaxial deformation was applied along the long axis of the molecular models, N-crosslinks aligned rapidly at lower strains followed by C-crosslinks more gradually at higher strains, leading to a two-stage crosslink recruitment. Then when comparing the influence of different enzymatic crosslinks, significant differences were observed for the high-strain elastic moduli of our microfibril unit models, namely and in increasing order, uncrosslinked, immature crosslinked (HLKNL and deH-HLNL), mature HHL-crosslinked, and mature PYD-crosslinked. At the fibril level, our low- and high-strain elastic moduli were in good agreement with some literature data, but in over-estimation of several other literature reports. Future work will seek to address simplifications and limitations in our modeling approach. A model such as this, accounting for different enzymatic crosslink types, may allow for the prediction of the mechanics of collagen fibrils and collagenous tissues, in representation of healthy and diseased states. / Ph. D.

Le terme invadosome regroupe à la fois les podosomes, dans les cellules normales, et les invadopodes, dans les cellules transformées par l’oncogène Src et les cellules cancéreuses. Ces structures sont capables d’interagir avec et de dégrader la matrice extracellulaire (MEC). Ils sont aussi considérés comme des méchanosenseurs car ils sont capables de sentir la rigidité et la nature de la MEC. Mon travail de thèse s’est focalisé sur l’impact du microenvironnement matriciel sur la formation et l’activité des invadosomes. Au cours d’une première étude, nous avons démontré que les cellules endothéliales microvasculaires forment de façon constitutive des podosomes. L’utilisation de matrices de rigidités contrôlées, a permis la mise en évidence d’une corrélation entre l’augmentation de la rigidité augmentait et le nombre de cellules formant des podosomes ainsi que la taille de ces structures. En plus de la rigidité, d’autres propriétés de la MEC, telles que sa composition moléculaire et son organisation pourraient influencer la formation des invadosomes. Dans une seconde et troisième étude, nous avons pu montrer que seul le collagène fibrillaire de type I était capable d’induire la formation de microdomaines d’actine linéaires qui présentent comme les invadosomes, la capacité de dégrader la MEC. Au vu de leur morphologie originale, nous avons nommés ces structures des invadosomes linéaires (LIs). De façon intéressante, nous avons pu établir que la formation et l’activité de dégradation des LIs étaient indépendantes des intégrines β1 et β3. Au contraire, nous avons démontré que les récepteurs à domaine discoïdine (DDRs) contrôlent la formation et l’activité des LIs. De plus, les voies de signalisation classiques associées aux invadosomes classiques ne sont pas impliquées dans la formation des LIs. Une analyse par spectrométrie de masse des interactants de DDR1 dans un contexte collagène de type I fibrilllaire a permis de mettre en évidence des régulateurs clés et de révéler une voie de signalisation potentiellement impliquée dans la formation des LIs.Ainsi, ce travail de thèse a permis d’identifier la rigidité de la matrice comme un inducteur majeur des podosomes mais aussi la capacité intrinsèque des cellules microvasculaires à former ces structures. De plus, nous avons identifié un nouveau type d’invadosome, les LIs, qui sont associés à un nouveau type de récepteur concernant les invadosomes, les DDRs. / Invadosome is a global term including podosome, found in normal cells, and invadopodia observed in Src-transformed and cancer cells. These structures are specialized cell-matrix contacts able to interact with and degrade the extracellular matrix (ECM). They are considered as mechanosensors as they are able to sense the strength, the nature of the extracellular matrix. My PhD work essentially focuses on the understanding of how matrix microenvironment impacts on invadosome formation and activity. In a first study, we demonstrated that microvascular cell types constitutively form podosomes. Thus, using matrices of controlled rigidity, we found that an increase of stiffness was associated with an enhancement in the number of cells forming podosomes and podosome size. In addition to the matrix rigidity, other microenvironment properties, such as the molecular composition and the organization of the matrix are expected to influence the formation of invadosome.In a second and a third part of this wok, we show that fibrillar type I collagen induce the formation of linear actin microdomains which exhibit invadosome characteristics. In view of their original architecture, we named these new structures, linear invadosomes (LIs).Interestingly, we show that the formation and degradation activity of LIs are independent of β1 and β3-integrins but required discoidin domain receptors (DDRs). Moreover, all the signalling pathways known to induce classical invadosome are not required for the LIs induction. A mass spectrometry analysis of DDR1 partners emphasized key regulators and these results highlight a new potential signalling pathway involved in LIs formation. This work allowed us identifying matrix stiffness as a major inducer of podosomes but also the intrinsic capacity of microvascular cells to form these structures. Moreover, we identify a new type of invadosome, the Linear Invadosome associated with DDRs receptors.

Low molecular weight gelators (LMWGs) are small molecules that are capable of entrapping solvents to form a gel in organic solvents or aqueous solution. These compounds rely solely on noncovalent forces to form the fibrous networks necessary to entrap a variety of solvents. The organogels and hydrogels thus formed could have applications in a variety of fields from environmental to biological to medicinal. Carbohydrates are ideal starting materials to synthesize LMWGs, because of their natural abundance, dense chirality, and biocompatibility. D-Glucose is the most common monosaccharide and D-glucosamine is isolated from natural sources, such as crab shells. Several series of compounds were synthesized using compounds 1-3 as the starting materials. These include esters, carbamates, amides, and ureas. The structure and gelation relationship was analyzed to obtain guidelines for designing new LMWGs. Compound 1 is a simple derivative of D-glucose and its terminal alkynyl esters and saturated carbamates are effective gelators. Compound 2 is a simple derivative of D-glucosamine and its amide and urea derivatives are also effective gelators. Compound 3 is formed from the deoxygenation of D-glucose. 1OOHOOCH3OHOPh2OOHOOCH3NH2OPh3OOHOOHOPh The design, synthesis and gelation properties of several classes of sugar based low molecular organo/hydrogelators will be discussed in this thesis in chapters 2, 3, and 4. After obtaining highly effective organo/hydrogelators, potential applications of these novel molecular systems can be explored. Some preliminary study on using one of the gelator in enzyme assay has shown that it is possible to utilize the hydrogels to immobilize enzymes. However, future research can explore further on the applications of these gelators.

Förster Resonance Energy Transfer (FRET)-based techniques are gaining an increasing importance in cell biology and cell-matrix adhesion studies because they allow both the detection of conformational changes of target proteins and their localisation in cells. Frequency Domain-Fluorescence Lifetime Microscopy (FD-FLIM) is currently considered one of the most reliable methods to measure FRET in live cells. However, due to its dependence on many technical prerequisites, its use is not yet widespread. The purpose of this work was to first establish FD-FLIM measurements of FRET on a new FD-FLIM microscope module. Then we aimed to apply FD-FLIM-FRET measurements to the study of conformational changes of the cell matrix-adhesion proteins vinculin and integrin and of the growth factor receptor Tie-2. In the first part of the work, published FRET probes including distance-sensors and two sets of vinculin-based probes were extensively tested with FD-FLIM, sensitised emission and ratiometric FRET. FD-FLIM was shown to be the most accurate method in approximating molecular distances between fluorophores. Moreover this study unveiled specific caveats associated with both existing vinculin FRET probes. FD-FLIM was then used to study conformational changes of the extracellular matrix receptor alphavβ3 integrin and of the angiopoietin receptor Tie-2 using specific FRET probes designed by us. While data showed that the alphav-integrin-FRET probe localised to adhesion sites, more experiments will be required to evaluate its full functionality. The Tie-2-FRET probe was fully functional and, upon ligand binding, allowed the detection of a bending movement of the extracellular domain towards the cell membrane. Finally, a combination of FRET, immunofluorescence and tension release experiments were used to show that intracellular tension is not required to maintain integrins in their activated conformation. However, intracellular tension is required to recruit other key proteins such as vinculin, talin and tensin to adhesions sites. Overall this work demonstrates the importance of FD-FLIM-FRET as a tool to investigate conformational changes of adhesion proteins and transmembrane receptors within the cell environment.

O câncer de mama é o tipo mais frequente de neoplasia maligna entre as mulheres, com a incidência de mais de um milhão de novos casos no mundo, por ano. O gene TP53 é responsável por regular o destino da célula em resposta a estresses genotóxicos e não genotóxicos. Mutações somáticas neste gene são encontradas em, aproximadamente, 50% dos carcinomas humanos. No câncer de mama, a frequência das mutações no TP53 é em torno de 20 a 50%, sendo a alteração mais encontrada. Estas mutações podem alterar a conformação da proteína, prejudicando sua função de ativadora da transcrição de genes alvo e, pode levar a p53 a apresentar tendência à agregação. Neste trabalho, foi realizado a análise da presença de mutações nos exons 5 a 10 do gene TP53, em biópsias de câncer de mama, de mulheres residentes na cidade do Rio de Janeiro. Após a identificação das amostras com mutação em TP53, algumas foram selecionadas para se verificar o comportamento das diversas mutantes quanto à formação de agregados de p53, em cortes histológicos dos tumores de mama. Foram utilizados os anticorpos A11 e DO1, que reconhecem oligômeros pré-fibrilares e a p53 mutante ou selvagem, respectivamente. Para tanto, foi utilizado o ensaio de co-localização por imunofluorescência, utilizando microscópio confocal para a observação dos resultados. Mutações no gene TP53 foram detectadas em 19% dos casos analisados de câncer de mama sendo 88,2% do tipo &#8213;missense&#8214;. Estas mutações, em tumores do tipo carcinoma ductal infiltrante (CDI), estavam associadas a um estágio mais tardio e agressivo de câncer, representado pelo Grau III de Elston (p< 0,0001). Também foi observada relação positiva dos tumores com mutações e o acúmulo da p53 (p= 0,0184). Além disso, foi observado um padrão diferente das mutantes de p53 quanto à tendência a agregação, sendo as mutantes R273H e P278A as que apresentaram uma maior agregação. Assim, a agregação da p53 mutante observada &#8213;in vivo&#8214; e descrita nesta dissertação, parece depender do tipo de mutação observada nos casos de câncer de mama. / Breast cancer is the most frequent cancer in women, with 1 million of new cases in the world each year. The TP53 gene is responsible for the regulation of the cells fate in response to genotoxic and non-genotoxic stress. Somatic TP53 mutations are found in, approximately, 50% of human cancers. In breast cancer, TP53 mutation is the most frequent genetic alteration, being present in 20 to 50% of cases. These mutations may change the protein conformation, impairing the transcription of target genes, and may lead the mutant p53 to aggregate. In this study, we analyzed the presence of mutations in exons 5-10 of TP53 in breast cancer biopsies of women resident in the metropolitan area of Rio de Janeiro. After the determination of samples with mutation, some of them were selected to investigate the behavior of different mutants in the formation of p53 aggregates in tumors using A11 and DO-1 antibodies, which recognizes pre-fibrillar oligomers and mutant or wild-type p53, respectively. So, we utilized a immunofluorescence co-localization assay using confocal microscope. TP53 mutations were detected in 19% of the breast cancer cases, with 88.2% of missense type. These mutations, in tumors classified as infiltrating ductal carcinoma (CDI), were associated with a later and aggressive stage of cancer, represented by Elstons Grade III (p< 0.0001). A relation was also observed between these mutations and p53 accumulation (p= 0.0184). Furthermore, a different pattern of p53 mutants for the tendency to aggregate was observed and we detected that the mutants R273H and P278A showed a greater aggregation. Thus, the mutant p53 aggregation observed in vivo and described in this study, seems to depend on the type of mutation found in breast cancer cases.

O câncer de mama é o tipo mais frequente de neoplasia maligna entre as mulheres, com a incidência de mais de um milhão de novos casos no mundo, por ano. O gene TP53 é responsável por regular o destino da célula em resposta a estresses genotóxicos e não genotóxicos. Mutações somáticas neste gene são encontradas em, aproximadamente, 50% dos carcinomas humanos. No câncer de mama, a frequência das mutações no TP53 é em torno de 20 a 50%, sendo a alteração mais encontrada. Estas mutações podem alterar a conformação da proteína, prejudicando sua função de ativadora da transcrição de genes alvo e, pode levar a p53 a apresentar tendência à agregação. Neste trabalho, foi realizado a análise da presença de mutações nos exons 5 a 10 do gene TP53, em biópsias de câncer de mama, de mulheres residentes na cidade do Rio de Janeiro. Após a identificação das amostras com mutação em TP53, algumas foram selecionadas para se verificar o comportamento das diversas mutantes quanto à formação de agregados de p53, em cortes histológicos dos tumores de mama. Foram utilizados os anticorpos A11 e DO1, que reconhecem oligômeros pré-fibrilares e a p53 mutante ou selvagem, respectivamente. Para tanto, foi utilizado o ensaio de co-localização por imunofluorescência, utilizando microscópio confocal para a observação dos resultados. Mutações no gene TP53 foram detectadas em 19% dos casos analisados de câncer de mama sendo 88,2% do tipo &#8213;missense&#8214;. Estas mutações, em tumores do tipo carcinoma ductal infiltrante (CDI), estavam associadas a um estágio mais tardio e agressivo de câncer, representado pelo Grau III de Elston (p< 0,0001). Também foi observada relação positiva dos tumores com mutações e o acúmulo da p53 (p= 0,0184). Além disso, foi observado um padrão diferente das mutantes de p53 quanto à tendência a agregação, sendo as mutantes R273H e P278A as que apresentaram uma maior agregação. Assim, a agregação da p53 mutante observada &#8213;in vivo&#8214; e descrita nesta dissertação, parece depender do tipo de mutação observada nos casos de câncer de mama. / Breast cancer is the most frequent cancer in women, with 1 million of new cases in the world each year. The TP53 gene is responsible for the regulation of the cells fate in response to genotoxic and non-genotoxic stress. Somatic TP53 mutations are found in, approximately, 50% of human cancers. In breast cancer, TP53 mutation is the most frequent genetic alteration, being present in 20 to 50% of cases. These mutations may change the protein conformation, impairing the transcription of target genes, and may lead the mutant p53 to aggregate. In this study, we analyzed the presence of mutations in exons 5-10 of TP53 in breast cancer biopsies of women resident in the metropolitan area of Rio de Janeiro. After the determination of samples with mutation, some of them were selected to investigate the behavior of different mutants in the formation of p53 aggregates in tumors using A11 and DO-1 antibodies, which recognizes pre-fibrillar oligomers and mutant or wild-type p53, respectively. So, we utilized a immunofluorescence co-localization assay using confocal microscope. TP53 mutations were detected in 19% of the breast cancer cases, with 88.2% of missense type. These mutations, in tumors classified as infiltrating ductal carcinoma (CDI), were associated with a later and aggressive stage of cancer, represented by Elstons Grade III (p< 0.0001). A relation was also observed between these mutations and p53 accumulation (p= 0.0184). Furthermore, a different pattern of p53 mutants for the tendency to aggregate was observed and we detected that the mutants R273H and P278A showed a greater aggregation. Thus, the mutant p53 aggregation observed in vivo and described in this study, seems to depend on the type of mutation found in breast cancer cases.

A doença pulmonar obstrutiva crônica (DPOC) é caracterizada por inflamação crônica e alterações estruturais que levam a obstrução das pequenas vias aéreas e destruição do parênquima alveolar. A composição da matriz extracelular (MEC) nos pulmões tem um importante papel em prover e sustentar a arquitetura pulmonar. No entanto, não há uma descrição abrangente da composição da matriz extracelular no trato respiratório de indivíduos portadores de DPOC. No presente estudo investigou-se a composição da MEC das vias aéreas grandes (VAG), pequenas (VAP) e do parênquima pulmonar de pacientes com DPOC. Utilizando imunohistoquímica e análise de imagem analisou-se a área fracionada de fibras elásticas, colágenos I, III e IV, versicam, decorina, biglicano, lumicam, fibronectina e tenascina nas VAG, VAP e no parênquima peribrônquico e distal de 26 indivíduos com DPOC e comparou-se à área fracionada nos pulmões de 26 fumantes sem DPOC e 16 indivíduos não fumantes. A área fracionada de fibras elásticas foi significante maior no grupo de fumantes não obstruídos em comparação com os demais grupos, em todos os compartimentos analisados. Houve menor expressão de colágeno I na camada interna das VAG e nas camadas interna, muscular e externa das VAP dos indivíduos com DPOC e na camada externa das VAP dos fumantes não obstruídos quando comparados ao grupo controle. A área fracionada de versicam mostrou-se menor apenas no parênquima distal do grupo DPOC comparado ao grupo controle. O estudo da matriz de glicoproteínas mostrou maior área fracionada de fibronectina nas camadas interna, muscular e externa das VAP dos indivíduos com DPOC comparados aos demais grupos, assim como maior área fracionada de tenascina foi observado na membrana basal das VAG e na camada interna das VAP do grupo DPOC comparados aos controles. Além disso, a composição da MEC correlacionou-se com valores funcionais, como o VEF1 (% predito). A partir desses resultados, concluímos que a DPOC é caracterizada por complexas alterações nas principais proteínas estruturais nas pequenas e grandes vias aéreas. Tais alterações podem contribuir para a lesão tecidual persistente e com a obstrução ao fluxo aéreo observado na DPOC / COPD is characterized by chronic inflammation and structural alterations leading to small airway obstruction and to destruction of the lung parenchyma. The extracellular matrix (ECM) composition of the lungs has an important role in determining airway structure. However, there are no comprehensive descriptions of the ECM composition along the respiratory tract in COPD patients. We postulated that the ECM composition in large and small airways and in lung parenchyma of COPD patients differs from that observed in smoking and non-smoking controls. Using immunohistochemistry and image analysis, fractional areas of elastic fibers, type-I, -III and IV collagen, the proteoglycans versican, decorin, biglycan and lumican; fibronectin and tenascin were quantified in the large (LA) and small airways (SA), in peribronchiolar (PP) and distal parenchyma (DP) of 26 COPD patients and compared to 26 smokers without COPD and 16 non-smoking controls. The fractional area of elastic fibers was higher in non-obstructed smokers than in COPD and non-smoking controls subjects, in all lung compartments. Type-I collagen fractional area was lower in the inner layer of LA and in the inner, muscle and outer layer (OL) of SA of COPD patients and in the OL of SA of non-obstructed smokers when compared to non-smoking controls. The versican fractional area was lower in DP of COPD patients than non-smokers. Fibronectin fractional área was higher in the inner, muscle and outer layer of SA of COPD patients compared to non-smokers. Tenascin fractional area was higher in the subepithelial area of LA and inner layer of SA of COPD when compared to non-smoking controls. Furthermore, ECM composition correlated with FEV1% predicted. Architectural alterations due to an altered ECM composition in COPD are likely to contribute to the persistent tissue injury and to the airflow obstruction characteristic of this disease

A doença pulmonar obstrutiva crônica (DPOC) é caracterizada por inflamação crônica e alterações estruturais que levam a obstrução das pequenas vias aéreas e destruição do parênquima alveolar. A composição da matriz extracelular (MEC) nos pulmões tem um importante papel em prover e sustentar a arquitetura pulmonar. No entanto, não há uma descrição abrangente da composição da matriz extracelular no trato respiratório de indivíduos portadores de DPOC. No presente estudo investigou-se a composição da MEC das vias aéreas grandes (VAG), pequenas (VAP) e do parênquima pulmonar de pacientes com DPOC. Utilizando imunohistoquímica e análise de imagem analisou-se a área fracionada de fibras elásticas, colágenos I, III e IV, versicam, decorina, biglicano, lumicam, fibronectina e tenascina nas VAG, VAP e no parênquima peribrônquico e distal de 26 indivíduos com DPOC e comparou-se à área fracionada nos pulmões de 26 fumantes sem DPOC e 16 indivíduos não fumantes. A área fracionada de fibras elásticas foi significante maior no grupo de fumantes não obstruídos em comparação com os demais grupos, em todos os compartimentos analisados. Houve menor expressão de colágeno I na camada interna das VAG e nas camadas interna, muscular e externa das VAP dos indivíduos com DPOC e na camada externa das VAP dos fumantes não obstruídos quando comparados ao grupo controle. A área fracionada de versicam mostrou-se menor apenas no parênquima distal do grupo DPOC comparado ao grupo controle. O estudo da matriz de glicoproteínas mostrou maior área fracionada de fibronectina nas camadas interna, muscular e externa das VAP dos indivíduos com DPOC comparados aos demais grupos, assim como maior área fracionada de tenascina foi observado na membrana basal das VAG e na camada interna das VAP do grupo DPOC comparados aos controles. Além disso, a composição da MEC correlacionou-se com valores funcionais, como o VEF1 (% predito). A partir desses resultados, concluímos que a DPOC é caracterizada por complexas alterações nas principais proteínas estruturais nas pequenas e grandes vias aéreas. Tais alterações podem contribuir para a lesão tecidual persistente e com a obstrução ao fluxo aéreo observado na DPOC / COPD is characterized by chronic inflammation and structural alterations leading to small airway obstruction and to destruction of the lung parenchyma. The extracellular matrix (ECM) composition of the lungs has an important role in determining airway structure. However, there are no comprehensive descriptions of the ECM composition along the respiratory tract in COPD patients. We postulated that the ECM composition in large and small airways and in lung parenchyma of COPD patients differs from that observed in smoking and non-smoking controls. Using immunohistochemistry and image analysis, fractional areas of elastic fibers, type-I, -III and IV collagen, the proteoglycans versican, decorin, biglycan and lumican; fibronectin and tenascin were quantified in the large (LA) and small airways (SA), in peribronchiolar (PP) and distal parenchyma (DP) of 26 COPD patients and compared to 26 smokers without COPD and 16 non-smoking controls. The fractional area of elastic fibers was higher in non-obstructed smokers than in COPD and non-smoking controls subjects, in all lung compartments. Type-I collagen fractional area was lower in the inner layer of LA and in the inner, muscle and outer layer (OL) of SA of COPD patients and in the OL of SA of non-obstructed smokers when compared to non-smoking controls. The versican fractional area was lower in DP of COPD patients than non-smokers. Fibronectin fractional área was higher in the inner, muscle and outer layer of SA of COPD patients compared to non-smokers. Tenascin fractional area was higher in the subepithelial area of LA and inner layer of SA of COPD when compared to non-smoking controls. Furthermore, ECM composition correlated with FEV1% predicted. Architectural alterations due to an altered ECM composition in COPD are likely to contribute to the persistent tissue injury and to the airflow obstruction characteristic of this disease

Descrevemos modelo original de sinovite experimental em coelhos imunizados com colágeno V com escasso processo inflamatório, intenso remodelamento matricial e vasculite. Analise morfológica e bioquímica foi realizada em coelhas Nova Zelândia (N=20) imunizadas com colágeno do tipo V, comparadas com controles. Foi observado o aumento dos colágenos I, III e V, oclusão do lúmen vascular e escasso processo inflamatório. A análise bioquímica confirmou a fibrose com aumento da síntese de colágeno. Nós postulamos que as alterações sinoviais descritas neste modelo foram conseqüência das particularidades do colágeno V, que promove manifestações imunológicas e clínicas semelhantes à esclerodermia / We described an original model of experimental synovitis in rabbits immunized with collagen V with scant cellular infiltration, intense matrix remodeling and vasculitis. Morphological and biochemical analysis were realized in New Zealand female rabbits (N=20) immunization with type V collagen, compared with control rabbits. It was observed increase of collagen I, III and V, vascular lumen occlusion and scant inflammatory process. Biochemical analysis confirmed the fibrosis with increased synthesis of collagen. We postulate that synovial changes described in this model are consequence of collagen V particularities, which promotes immunologic and clinical manifestations similar to scleroderma

Researches on biological adhesive systems in nature have changed a perspective view on adhesion that it is not only the area of surface chemistry, but also mechanics of interfacial geometry which can significantly effect on fracture strength and load distribution on the contact interface. Various synthetic fibrillar adhesives in previous works have shown enhanced interfacial bond strength with the capacity of adhesion control by exploiting mechanical deformation of the elastomeric fibrillar structures inspired by geckos. However, control of the interfacial load distribution has been focused on the size of micro-contact with single or a few of micro-/nano-fibers on planar surface, and not for a large contact area on complex three-dimensional (3D) surfaces. This thesis work aims at investigating principles of the interfacial load distribution control in multi-scale, ranging from micro-contact with single micro-fiber to a centimeter-scale contact with a membrane-backed micro-fiber array on non-planar 3D surfaces. The findings are also applied for developing a soft robotic gripper capable of grasping a wide range of complex objects in size, shape, and number, expanding the area of practical applications for bio-inspired adhesives in transfer printing, robotic manipulators, and mobile robots. This paper comprises three main works. First, we investigate the effect of tip-shapes on the interfacial load sharing of mushroom-shaped micro-fibrillar adhesives with precisely defined tipgeometries using high resolution 3D nano-fabrication technique. For a large area of non-planar contact interface, we fabricate fibrillar adhesives on a membrane (FAM) by integrating micro-fibers with a soft backing, which enables robust and controllable adhesion on 3D surfaces. Picking and releasing mechanism for the maximal controllability in adhesion are discussed. Finally, we propose a soft robotic architecture which can control the interfacial load distribution for the FAM on 3D surfaces, solving an inherit dilemma between conformability and high fracture strength with the equal load sharing on complex non-planar 3D surfaces.