Personne : Guillemette, Maxime.
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Département de physique, de génie physique et d'optique, Faculté des sciences et de génie, Université Laval
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- PublicationAccès libreAlignment of cells and extracellular matrix within tissue-engineered substitutes(Intech, 2013-01-01) Guillemette, Maxime.; Germain, Lucie; Bourget, Jean-Michel; Veres, Teodor; Auger, François A.Most of the cells in our body are in direct contact with extracellular matrix (ECM) components which constitute a complex network of nano-scale proteins and glycosaminoglycans. Those cells constantly remodel the ECM by different processes. They build it by secreting different proteins such as collagen, proteoglycans, laminins or degrade it by producing factors such as matrix metalloproteinase (MMP). Cells interact with the ECM via specific receptors, the integrins . They also organize this matrix, guided by different stimuli, to generate patterns, essential for tissue and organ functions. Reciprocally, cells are guided by the ECM, they modify their morphology and phenotype depending on the protein types and organization via bidirectional integrin signaling [2-4]. In the growing field of tissue engineering , control of these aspects are of the utmost importance to create constructs that closely mimic native tissues. To do so, we must take into account the composition of the scaffold (synthetic, natural, biodegradable or not), its organization and the dimension of the structure. The particular alignment patterns of ECM and cells observed in tissues and organs such as the corneal stroma, vascular smooth muscle cells (SMCs), tendons, bones and skeletal muscles are crucial for organ function. SMCs express contraction proteins such as alpha-smoothmuscle (SM)-actin, desmin and myosin  that are essential for cell contraction . To result in vessel contraction, the cells and ECM need to be organized in such a way that most cells are elongated in the same axis. For tubular vascular constructs, it is suitable that SMCs align in the circumferential direction, as they do in vivo [7, 8]. Another striking example of alignment is skeletal muscle cells that form long polynuclear cells, all elongated in the same axis. Each cell generates a weak and short contraction pulse but collectively, it results in a strong, long and sustained contraction of the muscle and, in term, a displacement of the member. In the corneal stroma, the particular arrangement of the corneal fibroblasts (keratocytes) and ECM is essential to keep the transparency of this tissue [9-13]. Tendons also present a peculiar matrix alignment relative to the muscle axis. It gives a substantial resistance and exceptional mechanical properties to the tissue in that axis [14, 15]. Intervertebral discs , cartilage , dental enamel , and basement membrane of epithelium are other examples of tissues/organs that present peculiar cell and matrix organization. By reproducing and controlling those alignment patterns within tissue-engineered substitutes, a more physiological representation of human tissues could be achieved. Taking into account the importance of cell microenvironment on the functionality of tissue engineered organ substitutes, one can assume the importance of being able to customise the 3D structure of the biomaterial or scaffold supporting cell growth. To do so, some methods have been developed and most of them rely on topographic or contact guidance. This is the phenomenon by which cells elongate and migrate in the same axis as the ECM. Topographic guidance was so termed by Curtis and Clark  to include cell shape, orientation and movement in the concept of contact guidance described by Harrison  and implemented by Weiss [21, 22]. Therefore, if one can achieve ECM alignment, cells will follow the same pattern. Inversely, if cells are aligned on a patterned culture plate, the end result would be aligned ECM deposition . The specific property of tissues or materials that present a variation in their mechanical and structural properties in different axis is called anisotropy. This property can be evaluated either by birefringence measurements [24, 25], mechanical testing in different axis , immunological staining of collagen or actin filaments  or direct visualisation of collagen fibrils using their self-fluorescence around 488 nm [27, 28]. Several techniques have been recently developed to mimic the specific alignment of cells within tissues to produce more physiologically relevant constructs. In this chapter, we will describe five different techniques, collagen gel compaction, electromagnetic field, electro‐spinning of nanofibers, mechanical stimulation and microstructured culture plates.
- PublicationRestreintTissue engineering of human cornea(CRC Press, 2014-03-27) Guillemette, Maxime.; Giasson, Claude-J.; Guérin, Sylvain; Germain, Lucie; Auger, François A.; Gaudreault, Manon.; Proulx, Stéphanie; Carrier, Patrick; Chirila, TraianThe cornea is a well-organized tissue composed of three cell types (epithelial, stromal and endothelial cells), each having an important role for its functionality. This chapter will address different tissue engineering approaches to the reconstruction of either partial or full-thickness living corneal substitutes that can be used either as in vitro models for woundhealing studies, or in vivo, eventually replacing the donor cornea for transplantation in humans. Isolation of the proper cells, followed by appropriate culture conditions, and assembly into a three-dimensional tissue construct, are the first steps required for producing a functional corneal substitute.