No description available.

The alveolar epithelium is a key functional component of the air-blood barrier in the lung. Comprised of two morphologically distinct cell types, alveolar epithelial type I (ATI) and type II (ATII) cells, effective repair of the alveolar epithelial barrier following injury appears to be an important determinant of clinical outcome. The prevailing view suggests this repair is achieved by the proliferation of ATII cells and the transdifferentiation of ATII cells into ATI cells. Supplemental oxygen and mechanical ventilation are key therapeutic interventions in the supportive treatment of respiratory failure following lung injury, but the effects of hyperoxia and mechanical deformation in the injured lung, and on alveolar epithelial repair in particular, are largely unknown. The clinical impression however, is that poor outcome is associated with exposure of injured (repairing) epithelium to such iatrogenic ‘hits’. This thesis describes studies investigating the hypothesis that hyperoxia & mechanical deformation inhibit normal epithelial repair. The in vitro data presented demonstrate that hyperoxia reversibly inhibits the transdifferentiation of ATII-like cells into ATI-like cells with time in culture. Whilst confirming that hyperoxia is injurious to alveolar epithelial cells, these data further suggest the ATII cell population harbours a subpopulation of cells resistant to hyperoxia-induced injury. This subpopulation of cells appears to generate fewer reactive oxygen species and express lower levels of the zonula adherens protein E-cadherin. Using a panel of antibodies to ATI (RTI40) and ATII (MMC4 & RTII70) cell-selective proteins, the effect of hyperoxia on the phenotype of the alveolar epithelium in a rat model of resolving S. aureus-induced lung injury was investigated. These in vivo studies support the view that, under normoxic conditions, alveolar epithelial repair occurs through ATII cell proliferation & transdifferentiation of ATII cells into ATI cells, with transdifferentiation occurring via a novel intermediate (MMC4/RTI40-coexpressing) immunophenotype. However, in S. aureus-injured lungs exposed to hyperoxia, the resolution of ATII cell hyperplasia was impaired, with an increase in ATII cell-staining membrane and a reduction in intermediate cell-staining membrane compared to injured lungs exposed to normoxia alone. As hyperoxia is pro-apoptotic and known to inhibit ATII cell proliferation, these data support the hypothesis that hyperoxia impairs normal epithelial repair by inhibiting the transdifferentiation of ATII cells into ATI cells in vivo. The effect of mechanical deformation on alveolar epithelial cells in culture was investigated by examining changes in cell viability following exposure of epithelial cell monolayers to quantified levels of cyclic equibiaxial mechanical strain. In the central region of monolayers, deformation-induced injury was a non-linear function of deformation magnitude, with significant injury occurring only following exposure to strains greater than those associated with inflation of the intact lung to total lung capacity. However, these studies demonstrate for the first time that different epithelial cell phenotypes within the same culture system have different sensitivities to deformation-induced injury, with spreading RTI40-expressing cells in the peripheral region of epithelial cell monolayers and in the region of ‘repairing’ wounds being injured even at physiological levels of mechanical strain. These findings are consistent with the hypothesis that alveolar epithelial cells in regions of epithelial repair are highly susceptible to deformation-induced injury.

The thymus is a complex cellular structure made up of several interdependent cell types and is the primary site for T cell development. A population of fetal thymic epithelial cells (TEC), marked by MTS20 and MTS24, when grafted in vivo can generate a functional thymus containing all thymic epithelial cells and is capab,e of supporting T cell differentiation. Further analysis using in vivo grafting experiments have determined the endoderm as the sole origin for all major thymic epithelial subsets. These findings suggest the possibility that a bipotent thmic epithelial progenitor cell (TEPC) gives rise to both cortical and medullary epithelial compartments. The first ai of this study was to address whether bipotent mouse TEPC give rise to both medullary and cortical epithelial cell populations and to begin to establish a model of TEC differentiation through ontogeny. Its second aim was to start to define condidtions for maintaining functionally undifferentiated RTEPC in vitro. Finally, as little is knowth about the genetic regulation of human thymic development and TEC differentiation, I hve used comparative analysis to investigate the similaarities in expression of key regulations of thymus development between human and mouse, which will aid in the translation of mouse thmic research to human. the main findings of this thesis are i) that a bipotent thymic epithelial progenitor cell population that contribute to both medullary and cortical epithelial cell compartments exists in vivo, but is at low frequency even by E12.5. ii) That a unipotent progenitor population committed to a cortical epithelial cell fate is also present as early as E11.5. iii) That E11.5 TEC can be propagated in vitro in semi defined conditions, but appear to revert ro an early endodermal phenotype on prolonged culture and iv) that the genetic program regulating thymus organogenesis appears to be conserved between mouse and human. In addition I have defined the exact time of haematopoietic cell entry into the human thymus, and the time of entry of mesenchymal cells and the mesenchymal distribution pattern throughout human thymic ontogeny. I also establish the time of onset of differentiation and maturation of hTEC, and using known mouse TEPC markets, show the existence of a population of Claudin4+Ueal+ hTEC at week 8, presumed equivalent to the precursor cells for the AIRE1+ subpopulation of medullary TEC in the mouse.

Orthokeratology (OK) is a specialty contact lens procedure that involves the overnight wear of lenses to reshape the corneal tissue resulting in clear vision upon lens removal. Currently it is the only way of achieving clear vision without having to wear spectacles or contact lenses during the day or undergoing refractive surgery. This thesis investigated the effects of this procedure on the corneal epithelium and the potential increase in risk of microbial infection in an animal model. The cat was first established as an appropriate animal model in a pilot study to examine OK epithelial changes. Initial findings of epithelial thickness changes similar to those found with human myopic and hyperopic OK clinical studies led to the further development of this animal model to better mimic human lens wear for the remaining studies undertaken. Histological studies were used to examine epithelial effects of overnight myopic OK lens wear. Repeatable and differential effects on epithelial thickness and morphology across the cornea were found, including thinning of the central and peripheral epithelium and thickening of the mid-peripheral epithelium. Central thinning was attributed to compression of cells and was less in overnight wear compared to continuous wear. Mid-peripheral thickening was due to increased cell layers and peripheral thinning was attributed to cellular compression. Recovery of epithelial morphologic and thickness changes commenced one day after ceasing lens wear and was complete within one week. Minimal changes to keratocyte populations in regions adjacent to epithelial thickness changes were found. Microbiological studies investigated the effect of epithelial changes on corneal susceptibility to bacterial infections by exposing OK-treated corneas to large amounts of Pseudomonas aeruginosa. The hypothesis that OK lenses increased susceptibility to infection (within the time tested) was rejected as no infections were produced in any animals (except the positive scratch control). Length of OK treatment, duration and quantity of bacterial exposure, lens wearing schedule and bacterial strain type did not affect susceptibility to infection. The epithelium is a major contributor to OK-induced corneal changes. These epithelial changes are reversible and do not appear to predispose to infection provided corneal integrity is maintained.

Thesis (Ph.D.)--Hong Kong University of Science and Technology, 2006. / Includes bibliographical references (leaves 134-151). Also available in electronic version.

published_or_final_version / Anatomy / Master / Master of Philosophy

No description available.

Orthokeratology (OK) is a specialty contact lens procedure that involves the overnight wear of lenses to reshape the corneal tissue resulting in clear vision upon lens removal. Currently it is the only way of achieving clear vision without having to wear spectacles or contact lenses during the day or undergoing refractive surgery. This thesis investigated the effects of this procedure on the corneal epithelium and the potential increase in risk of microbial infection in an animal model. The cat was first established as an appropriate animal model in a pilot study to examine OK epithelial changes. Initial findings of epithelial thickness changes similar to those found with human myopic and hyperopic OK clinical studies led to the further development of this animal model to better mimic human lens wear for the remaining studies undertaken. Histological studies were used to examine epithelial effects of overnight myopic OK lens wear. Repeatable and differential effects on epithelial thickness and morphology across the cornea were found, including thinning of the central and peripheral epithelium and thickening of the mid-peripheral epithelium. Central thinning was attributed to compression of cells and was less in overnight wear compared to continuous wear. Mid-peripheral thickening was due to increased cell layers and peripheral thinning was attributed to cellular compression. Recovery of epithelial morphologic and thickness changes commenced one day after ceasing lens wear and was complete within one week. Minimal changes to keratocyte populations in regions adjacent to epithelial thickness changes were found. Microbiological studies investigated the effect of epithelial changes on corneal susceptibility to bacterial infections by exposing OK-treated corneas to large amounts of Pseudomonas aeruginosa. The hypothesis that OK lenses increased susceptibility to infection (within the time tested) was rejected as no infections were produced in any animals (except the positive scratch control). Length of OK treatment, duration and quantity of bacterial exposure, lens wearing schedule and bacterial strain type did not affect susceptibility to infection. The epithelium is a major contributor to OK-induced corneal changes. These epithelial changes are reversible and do not appear to predispose to infection provided corneal integrity is maintained.

Thesis (M.D.S.)--University of Adelaide, Dept. of Dentistry, 1987. / Addenda inserted. Includes bibliographical references (leaves 125-145).

Thesis (Ph. D.)--University of Hong Kong, 2008. / Also available in print.