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Title: Cell-cell interactions and matrix deposition in tendon development in vitro and in vivo
Author: Fish, Rhiannon Susan
ISNI:       0000 0004 2747 6455
Awarding Body: Cardiff University
Current Institution: Cardiff University
Date of Award: 2005
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Tendons transmit tensile load from skeletal muscles to bone. They fundamentally consist of densely packed longitudinal collagen fibres separated by rows of tendon fibroblasts. The cells have extensive cell-cell contacts, longitudinally within the rows and between rows via cell processes. This thesis explores how cell-cell interactions and the cytoskeleton relate to matrix secretion and organisation in developing chicken tendons and cell cultures. In initial developmental stages, indicated by tenascin and type I collagen expression, cells were associated with cadherins, vinculin and connexin 32, and the presence of actin fibres. Subsequently, type III collagen delineated early fascicular structure, and later still connexin 43 appeared, relating to mechanical loading and formation of a cellular regulatory network. In culture, monolayers laid down minimal matrix, high density micromass cultures laid down more and pellet cultures produced an extensive matrix, although 75% of their collagen was lost to the medium. Micromasses and pellets expressed similar cytoskeletal and cell junctional markers to the in vivo tendons monolayers differed with early expression of connexin 43. TGF0, bFGF and PDGF singly or in combination had minor effects on collagen production and deposition and had no effect on cell junctional or cytoskeletal components. Finally, a novel suspension culture system was developed to allow large numbers of cells to interact and deposit matrix without interference from medium changes. After establishing that cells aggregated rapidly in suspension culture, suspensions were placed in dialysis tubes contained within 50ml sterile plastic tubes and cultured on rollers, with medium being changed only in the large tubes, leaving the cell suspensions undisturbed. This produced structures up to 3cm in length, with good cell and matrix organisation and seeming to incorporate more collagen into the extracellular matrix than pellet cultures. This could form the basis for a simple scaffold-free tissue engineering approach for tendons and ligaments.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available