Equine hyperelastosis cutis (HC or HERDA), a connective tissue disorder in American Quarter Horses, results in hyperelastic skin with poor wound healing. Similar conditions are found in many species and all forms display decreased skin tensile strength. Fibroblasts produce collagen and elastin fibers, forming networks, providing the dermis with strength, and elasticity. This study aims to carry out a 3-part evaluation between horse skin fibroblast (cells from horses affected with HERDA, cells from horses that are carriers of HERDA (recessive HERDA gene), and cells from horses that are normal (neither affected or carriers of HERDA); Studies include: 1. Cell proliferation assay 2. Apoptosis analysis of fibroblasts 3. Mechanobiology of stretched fibroblast. Studies have shown cellular deformation to have an overall effect on mechanical properties of healthy and unhealthy tissues. This investigation provides a micromechanical evaluation of HC/HERDA in an effort to quantify the cellular level differences between each condition.

Individual carbon nanotubes have been theoretically and experimentally proven to be the strongest and stiffest materials discovered to date with tensile strengths ranging from 1-5 TPa and elastic modulus values as high as 150 GPa. In this work, the recent development of continuous sheets of CNTs, produced by Nanocomp Technologies Inc ., are investigated for their potential as reinforcement in polymer matrix composite (PMC) materials. The potential of these nanotube-based PMC materials have been reported by researchers at Florida State University (FSU). Through the use of mechanical stretching procedures to increase the alignment of the nanotubes within the CNT sheets, the tensile strength and Young's modulus of the composites in the FSU study averaged 3081 MPa and 350 GPa, respectively. These values are for composites fabricated from 40% stretched CNT sheets and are 48% and 107% improvements over composites fabricated from the pristine, unstretched CNT sheets. However, the test specimens used in the FSU study consisted of a single CNT ply and each coupon was individually stretched and cured for testing. Therefore, the process used to generate the coupons which exhibited these high mechanical properties would be difficult to scale to a usable size for aerospace structural components. In the current study, a scalable process has been developed in which 2-ply, 3" x 3" panels of CNT and resin composites are fabricated. An apparatus and methodology for mechanically stretching the CNT sheets used in these composite panels has also been developed. After initial testing was conducted with the CNT composites and the coupons exhibited significant elongation at failure, along with the absence of a linear elastic region, conventional test standards for material testing were deemed impractical. For this reason, new mechanical testing methodologies have been developed to determine the mechanical properties of specific strength and specific modulus of CNT-polymer composites. In order to obtain the maximum benefits of a fiber in any matrix in terms of stiffness and strength, it is preferable to align the high strength and stiffness fibers in the direction of loading. Given that these CNT sheets essentially consist of billions of short, discontinuous CNTs of 2-3mmin length, the process of mechanical stretching is used in an attempt to align these tubes in the direction of the applied tensile load. Here we have explored methodologies for stretching, fabricating, and mechanical testing. Having identified a process which seems viable, an examination into the effect of the mechanical stretching to increase the alignment of the nanotubes within the CNT sheets, and thus to increase the material properties of the 2-ply composites constructed from them, is conducted. In order to correlate the enhancements in the mechanical properties with the increased alignment of the CNTs, polarized Raman spectroscopy techniques have been used. Lastly, Scanning Electron Microscopy (SEM) is used to examine the effect of stretching on the pristine CNT sheet, as well as examine the fracture surfaces of failed test coupons to better characterize the failure modes. In this report, polarized Raman spectroscopy has been used to confirm the enhancedalignment of nanotubes within the CNT sheets through the used of a nematic order parameter. Unstretched sheets exhibit an order parameter of 0.07 and 0.09 for untreated and Acetone treated sheets, respectively. Upon stretching the untreated sheets to 45%, the order parameter increases to 0.1409 and, when stretched to 30%, Acetone treated sheets have an order parameter of 0.1518. During the mechanical testing of 2-ply composites fabricated from stretched CNT sheets, the effect of this increased alignment is made apparent. Untreated CNT sheets are used to fabricate 2-ply composites after being stretched and are compared to baseline values of panels fabricated using sheets which are not stretched. In the panels fabricated with PEI resin and 43% stretched, untreated CNT sheets, a 137% increase in average specific strength and a 44% increase in average specific modulus over the baseline panel is observed. For panels fabricated with BMI and 33% stretched, untreated CNT sheets, a 169% increase in average specific strength and 105% increase in average specific modulus is observed when compared to the baseline panel. These increases are evidence for the potential of mechanical stretching to align the nanotubes within the CNT sheets and bolster the mechanical properties of resulting CNT-polymer composites. / Master of Science

La cicatrisation de ligaments est un processus complexe qui consiste en la prolifération et la migration cellulaire, ainsi que la synthèse de la matrice extracellulaire. Ce phénomène est influencé par des facteurs biochimiques et/ou environnementaux. Les propriétés spécifiques des cellules ligamentaires permettrent de répondre aux facteurs de croissance et aux contraintes mécaniques. Le but de nos travaux a été de définir les conditions de reconstruction in vitro d’un tissu ligamentaire. Nous avons tenu compte d’une part des paramètres biochimiques, d’autre part des paramètres mécaniques afin d’étudier l’ensemble des facteurs agissant sur la synthèse d’un nouveau tissu. Dans un premier temps, nous avons montré l’influence des facteurs de croissance, tels que le PDGF-AB et le TGF-b1 sur la prolifération et sur la synthèse de composants de la matrice extracellulaire (collagènes des types I et III). Il semble que ces deux facteurs de croissance aient une action, mais il n’agissent pas sur le même mode du point de vue temporel et mécanistique. Dans un deuxième temps, nous avons étudié l’effet de la contrainte mécanique spécifique du ligament, l’étirement. Nous avons montré que l’action des contraintes mécaniques est essentielle pour la différentiation des cellules cibles et leur évolution en cellules ligamentaires, en promouvant la synthèse de molécules spécifiques, telles que la ténascine, et les collagènes. Cependant, bien que ces résultats, restent insuffisants pour la reconstruction d’un bio tissu, ils nous ont néanmoins permis de définir les protocoles à suivre pour la suite de ces projets d’ingénierie tissulaire / The process of ligament healing is complex, which consists of the proliferation and the cellular migration, as well as the synthesis of the extra cellular matrix. This process is influenced by the biochemical and/or environmental factors. The specific properties of the ligament cell permit their responses to growth factors and to mechanical stress. The aim of our work was to define the conditions of in vitro reconstruction of ligament tissue. We take into account the biochemical, as well as the mechanical parameters in order to study the factors acting on the synthesis of a new tissue. Firstly, we investigated the influence of the growth factors such as PDGF-AB and TGF-b1 on the cell proliferation and the matrix synthesis (collagens types I and III) respectively. It seems that these two growth factors act on the different mode on the temporal and mechanistic aspects. Secondly, we studied the effect of the specific mechanical stress the stretching on the ligament. It was showed that the action of the mechanical stretching was essential for the differentiation of the target cells and their evolution in the ligament cell, by promoting the synthesis of the specific molecules, such as tenascin, and collagens. Although these results remain insufficient for the reconstruction of neotissue, they allowed us to define the protocols to be followed for the projects of tissue engineering