Physiological electric fields (EFs) exist in the vertebrate lens, but the importance of the endogenous EF is not well understood yet. In the present study, an applied EF to mimic endogenous EFs was applied to cultured lens epithelial cells (LECs) to investigate the effects of EFs on the behaviours of LECs and the underlying mechanisms. It was showed that LEG migration was directed and migration rate was increased in applied EFs, and serum or growth factors were required for these EF-induced cell responses. LECs elongated and re-oriented to lie perpendicular to field vector. Healing of LEG monolayer wounds was also influenced by EF polarity. EF exposure enhanced the activation of extracellular signal-regulated kinase (ERK) ½ and induced an asymmetric distribution of active ERK ½ in monolayer wounds. Mitogen-activated protein (MAP) kinase inhibitor U0126 inhibited the directed migration and reorientation of LECs in EFs and the healing of LEG monolayer wound, and U0126 also completely prevented activation of ERK ½ in LECs. It is suggested that MAP kinase signaling pathways were involved in the responses of LECs to EF stimulation. EF exposure also inhibited the proliferation of the LECs. Cell cycle analysis showed that EF exposure inhibited the Gl/S transition of the cell cycle progression in LECs, resulting in a Gl-block. The EF-induced down-regulated expression of Gl-specific cell cycle protein cyclin E and the up-regulated expression of cyclin-Cdk (cycle dependent kinase) complex inhibitor p27 kipl were accounted for the cell cycle arrest of LECs in EFs. This study implies that a physiological EF may be one of the guidance cues regulating LEG behaviours in vivo and applying EFs may be one way of controlling aberrant LEG behaviours in vitro and in vivo.

Thesis (Ph.D.)--Aberdeen University, 2008. / Title from web page (viewed on Apr. 20, 2009). Includes bibliographical references.

This work used <i>Pax6</i>^+/- mice as a model for Pax6-realted corneal diseases and assessed the roles of oxidative stress and epithelial injury in the aetiology of ARK. Histological investigation revealed epithelial lesions in <i>Pax</i>6^+/- corneas. Proteomic analysis demonstrated reduced levels of protective enzymes, transketolase, aldehyde dehydrogenase 3A1 and glutathione S-transferase α4, and cytoskeletal proteins. Keratin 5 and 12 in <i>Pax</i>6^+/- adult mouse corneas. These physical/structural and chemical defects imply that <i>Pax</i>6^+/- corneas may be in a chronic ‘stressed and wounded’ state. Using a DNPH/protein oxidation assay, <i>Pax</i>6^+/- corneal protein oxidation was found to be consistently higher than that in <i>wild-type</i> (WT), and to get worse with age, in parallel with the development of corneal opacity. H₂O₂ was used to induce oxidative stress in mouse corneas and this was found to activate the Ca²⁺ (protein kinase C/ phospholipase C) → p38/p42/p44 mitogen activated kinase signalling pathway. Oxidative stress-induced Pax6 exclusion form cell nucleus led to abnormal expression of non-corneal epithelial markers, indicating a metaplasia process that may cause normally transparent epithelial cells to become opaque. This report for the first time describes cytoskeleton architectures <i>in vivo</i> using flat-mount mouse corneal epithelial by fluorescent staining and confocal microscopy, which is potentially applicable to studies interested in cytoskeleton <i>in vivo</i>. Keratin, desmoplakin and actin cytoskeletal structures were found to be heterogeneous and defective in <i>Pax</i>6^+/- cells. Twenty-one hours after wounding WT corneal epithelia <i>in vivo</i>, healing cells developed desmoplakin and actin structural features, intercellular gaps, interdigitated filopodia-like processes and vesicles similar to the unscratched <i>Pax</i>6^+/- corneal epithelia. These data support the hypothesis of a ‘chronic wounded’ state in apparently uninjured <i>Pax</i>6^+/- corneal epithelia and reveal the cytoskeletal origins of poor adhesion and cellular structure.

Thesis (Ph.D.) -- University of Adelaide, Dept. of Surgery, 2000. / Two leaves of errata and addenda pasted into back pages. Bibliography: leaves 174-208.

Thesis (M.D.)--Dept. of Surgery, University of Adelaide, 2001? / Includes bibliography (leaves 191-219).

Non-healing wounds is a growing medical problem, often associated with pathological conditions such as diabetes and peripheral ischemia. A non-healing wound entails a large amount of suffering for the patient, and demands extensive health care resources. In this thesis, a new drug treatment paradigm for wound healing was developed by transforming Limosilactobacillus reuteri R2LC with a plasmid encoding CXCL12 (LB_CXCL12). The drug candidate was tested for safety and biological effects following topical administration to full thickness wounds in both mice and minipigs. In parallel, different techniques, including 2D and 3D measurements, planimetry, and ultrasound, for assessing wound healing were developed and evaluated.   Murine wounds treated with LB_CXCL12 demonstrated increased proliferation of dermal cells, and an increased density of macrophages of which a larger fraction expressed TGF-β. If macrophages were depleted prior to wounding, the accelerated effect on healing was abolished demonstrating a macrophage-dependent mechanism of action. Importantly, the LB_CXCL12 treatment also accelerated wound healing in mice with impaired healing as a result of hyperglycemia or peripheral ischemia, conditions that in humans are associated with development of non-healing wounds. Wounds in minipigs treated with the freeze-dried formulation of LB_CXCL12, upon resuscitation referred to as ILP100, showed accelerated healing both by increased granulation tissue formation and accelerated re-epithelialization. The treatment with ILP100 was well tolerated with no treatment-related deviations in haematology, urinalysis, and histopathology. Further, we found improved detection of thin layers if newly formed epithelial using planimetry and ultrasound compared to 2D photographs, whereas 3D scans accounting for surface curvatures yielded larger wound areas than 2D photographs of the same wounds.  Development of topical treatments for non-healing wounds are limited by the proteolytic environment of the wound that cause degradation of applied molecules. Our developed technology, a new-in-class candidate, overcomes this by continuous on-site delivery and increased bioavailability of CXCL12, resulting in prolonged instruction of local immune cells to stimulate wound healing.

Abstract Chronic, non-healing wounds constitute a massive financial burden on health care system. The healing processes of these wounds and their underlying pathology are only partly understood. In this study, important biological functions performed by Retinoic acid with its regulatory protein cellular retinoic acid binding protein 2 (CRABP2) were discussed. Possibly, these biological func-tions might be linked with chronic wound therapeutic by inducing antiproliferative activity of cells which leads to reduction in migration and growth rate of fibroblast during skin regeneration pro-cess in chronic wound healing. The aim of this study was to comparatively analyze the expression pattern of CRABP2 and P16 cyclic dependent kinase inhibitor in dermal fibroblasts at mRNA levels along with their morphological pattern, migration and growth rate. Fibroblasts were cultured and their morphology were observed by phase-contrast imaging. Difference in viability, migratory capacity was examined by Cell titer blue and scratch assay respectively and expression were meas-ured by polymerase chain reaction. Interestingly, the date revealed that morphology was altered and growth rate and migration velocity was significantly lower in chronic wound fibroblasts and senescent fibroblasts when compared with their control. Expression pattern revealed that CRABP2 was highly up-regulated only by senescent cells but not in chronic wound fibroblasts which point novel function for this protein in term of replicative senescence. However, P16 was not signifi-cantly altered among all fibroblasts which demands supplementary studies to conform the role of CRABP2 in fibroblast dysfunction and cellular senescence in chronic wounds.

Thesis (MMed) -- Stellenbosch University, 2010. / Bibliography / Chronic wounds afflict millions worldwide, incurring significant health care costs and chronic suffering. Clinicians are often unsure about treatment, resulting in poor outcomes. Objective To determine the scope of knowledge possessed by fifth year medical students, general practitioners (GP’s) and surgical registrars, concerning chronic wound management. Design Cross sectional study Methods Deans of eight South African medical schools received letters requesting information regarding time devoted to wound-care training. Knowledge-based questionnaires were distributed to final-year students at two universities, surgical registrars at three universities and general practitioners attending refresher courses. Result. Four medical schools replied, of whom only two offered formal teaching. 162 medical students, 45 GP’s and 47 surgical registrars completed questionnaires. The overall median (25th–75th percentiles) knowledge scores for registrars, GP’s and students were 65%;(55%–70%), 55%;(45%–65%) and 45%;(35%–50%) respectively. Whereas the scores of registrars and GP’s did not differ, the student scores were significantly less. Only 32% of registrars and 18% of GP’s attained scores of 70% or more. 96% considered training to be inadequate. Interest in wound-care was only mild to moderate, with more GP’s than registrars requesting literature. Conclusions Very little, if any training on chronic wounds is offered in South Africa. The levels of knowledge cannot be considered adequate for successful treatment, nor for teaching to undergraduates. This preliminary study cannot reflect the attitudes and knowledge throughout the country; however it is clear that there is a need for improved education about these conditions that have huge clinical and economic consequences.