The primary objective of my thesis was to investigate the effect of crimp-like fibrous scaffolds on bovine fibroblasts and to develop a scaffold for anterior cruciate ligament (ACL) tissue engineering. To achieve this objective, fibrous biodegradable polymeric scaffolds were fabricated, which upon relaxation developed a crimp-like structure, which resembled the crimp seen in native collagen. The understanding of the crimp mechanism allowed for controlling crimp-like patterns in various polymer fibre systems, and was determined to be due to residual stress coupled with an operating temperature (Top) above the glass transition temperature of the polymer (Tg). The benefit of crimp was evaluated by seeding fibroblasts on crimp-like fibres that were subjected to dynamic mechanical loading. The results showed a significant increase in extracellular matrix (ECM) accumulation by fibroblasts that experienced crimp unfolding. In addition, fibroblasts seeded on mechanically stimulated crimp-like fibrous scaffolds formed ECM bundles that resembled collagen fibre fascicles. Two separate studies were conducted to fabricate fibrous scaffolds with high modulus: one on thermoplastic polyesters and the other on a photocrosslinkable polyester. Of the thermoplastic polyesters investigated, poly(L-lactide-co-D,L-lactide) P(LLA-DLLA) exhibited the highest modulus, and was the most resistant to hydrolytic degradation. These fibres were placed in a heated aqueous environment to exhibit a crimp-like pattern similar to that of native collagen. Bovine fibroblasts were shown to attach, proliferate and deposit ECM on the surface of the P(LLA-DLLA) fibrous scaffolds. In addition, the deposited ECM appeared to be organized in distinctive bundles that resembled fascicles found in native ACL. However, upon crimp unfolding the crimp was not completely recovered. Photocrosslinkable poly(L-lactide-co-trimethylene carbonate cinnamate) P(LLA-TMC cinnamate) fibres in addition to supporting cell proliferation and ECM accumulation, completely recovered their crimp-like pattern, via [2 + 2] cycloaddition of the cinnamate groups. The recovery of crimp upon unfolding is a novel design feature incorporated into electrospun fibres as it innately mimics the function of collagen fibres found in the ACL. From the results obtained it is evident that crimp and its unfolding are key design features/conditioning techniques that need to be incorporated into fibrous scaffolds that possess high modulus, intended for ligament tissue engineering. / Thesis (Ph.D, Chemical Engineering) -- Queen's University, 2012-01-05 14:11:25.965

Made available in DSpace on 2016-12-08T15:59:06Z (GMT). No. of bitstreams: 1 Resumo Heiliane Fontana.pdf: 222602 bytes, checksum: 7b9629dbef6d399a129553dbf066ba26 (MD5) Previous issue date: 2015-05-05 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Embora as relações força-comprimento (F-C) e força-velocidade (F-V) de um músculo possam ser facilmente determinadas in-vitro, a determinação in-vivo é certamente mais complexa, e baseia-se frequentemente em medidas de torque e ângulo articular. Uma importante limitação dessa abordagem reside no fato de as mudanças de comprimento do fascículo muscular não se relacionarem de maneira simples às da unidade músculo-tendínea (UMT). Dessa forma, a investigação in-vivo da mecânica músculo-tendão torna-se necessária para a determinação da expressão dessas propriedades musculares no contexto do movimento humano. Neste trabalho, foi determinado, in-vivo, a relação F-C (Parte I) e a relação F-V (Parte II) nos níveis do fascículo e da UMT para contrações voluntárias do músculo vasto lateral (VL) em humanos. A relação F-C foi obtida para contrações máximas e submáximas e a relação F-V, para diferentes níveis de pré-contração do músculo. A fim de obter um melhor entendimento do fenômeno, o movimento de extensão de joelho foi analisado em diferentes amplitudes de acordo com características cinéticas e cinemáticas do mesmo. As coletas de dados envolveram medições sincronizadas de torque e ângulo de joelho, bem como arquitetura muscular e EMG do VL durante contrações isométricas (F-C) e isocinéticas (F-V) de extensores de joelho. Na Parte I, foi demonstrado que a quantidade de encurtamento dos fascículos em uma contração isométrica depende do comprimento da UMT/ ângulo articular, resultando em diferentes formatos para a relação F-C entre os níveis do fascículo e da UMT e entre contrações máximas e submáximas. As relações F-C para contrações submáximas foram determinadas a partir de níveis percentuais de força (como sugerido na literatura), mas, também, através de uma nova abordagem baseada em níveis percentuais de ativação (EMG). Com base na primeira abordagem, por definição, os picos de força para as relações F-C ocorrem em um comprimento muscular fixo, mas, devido a complacência da UMT, em comprimentos de fascículo maiores a medida que a força na UMT decresce. Contrariamente, na análise baseada em ativação, os picos de força ocorrem em um comprimento de fascículo constante, mas em comprimentos musculares menores a medida que a a força na UMT decresce. Este resultado sugere que o maior potencial de geração de força submáxima é obtido em comprimento de fascículo próximo ao definido como ótimo, ou seja, no comprimento onde a sobreposição dos miofilamentos é máxima. Na Parte II dessa tese, a análise da relação F-V evidenciou que a velocidade de encurtamento do fascículo durante a extensão isocinética de joelho pode ser menor, semelhante ou maior do que a da UMT dependendo da velocidade articular e de qual parte do movimento é analisada. A velocidade de encurtamento da UMT aumenta sistematicamente com a velocidade articular, enquanto que os fascículos podem permanecer com velocidades constantes para uma grande amplitude de velocidade articular. Os resultados apontam para a crítica natureza da interação entre ângulo articular, força/ativação muscular, elementos elásticos e mecânica dos fascículos musculares e auxiliam na superação das restrições encontradas quando se relaciona e se aplica conhecimento obtido nos diferentes níveis organizacionais dos músculos. / Although there are no major difficulties in obtaining the force-length (F-L) and force-velocity (F-V) relationships in skeletal muscles in-vitro, obtaining them in-vivo is certainly much more complex, and is often based on joint torque and angle. A primary problem is that fascicle length changes are not associated in a simple manner with changes in the length of the whole muscle-tendon unit (MTU). Therefore, investigation on whole muscle tendon mechanics in-vivo is needed in order to elucidate the link between these mechanical properties and the muscle performance in the context of human movement. In this thesis work, we determined the in-vivo F-L relationship (Part I) and F-V relationship (Part II) at the fascicle and MTU levels for voluntary contractions of the human vastus lateralis muscle (VL). F-L relationships were obtained for maximal and submaximal contractions, and F-V relationships for different pre-load conditions. In order to obtain a better understanding of the phenomena, different ranges of the knee extension movement were identified for their kinematic or kinetic properties and were analyzed separately. Data collection involved synchronized measurements of knee extensor torque, knee angle, VL EMG and muscle architecture (fascicle length and pennation angle) during isometric (F-L) and isokinetic (F-V) knee extensor contractions. In Part I, we demonstrated that fascicles undergo different amounts of shortening depending on MTU length/ joint angle, which results in different shapes of the fascicle and muscle F-L relationship, and produces different F-L shapes for maximal and submaximal levels of activation. The submaximal fascicle F-L relationships were determined based on percentages of force (as typically done in the literature), but novel to the literature, also based on percentages of activation (EMG). In the force-based analysis, the peak forces of F-L relationships, by definition, must occur at the same muscle length, but because of muscle-tendon compliance, it occurs at increasing fascicle lengths with decreasing levels of force. In contrast, in the activation-based analysis, the peak occurred at similar fascicle lengths, but because of muscle compliance, at decreasing muscle lengths with decreasing levels of force. This result suggests that, in the activation-based analysis, the greatest force generating potential always occurs close to optimal fascicle lengths; that is the length at which sarcomeres present maximum myofilament overlap. In Part II, analysis of the F-V relationship showed that fascicle shortening velocities during maximum isokinetic knee extension exercises can be slower, equal or faster than the corresponding MTU shortening velocities, depending on the knee extensor speed and depending on which part of the knee extension curve is analyzed. MTU shortening velocities increase systematically with increasing speeds of knee extension. However, this observation is not made for fascicle shortening velocities, which, depending on the range and conditions analyzed, remain about constant across a large range of knee extensor speeds. The results of this thesis point to the critical nature of the interaction between joint angle, muscle force/activation, series elastic compliance, and fascicle mechanics and help to overcome the difficulties in relating and applying knowledge obtained at the different organizational levels of muscles.

No description available.