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Biomedical subjects

Christopher J Murphy

Publications and source records attributed to Christopher J Murphy.

26 records · Page 2Linked to original sources
PubMed2003

Ultrastructural basement membrane topography of the bladder epithelium.

The basement membrane underlies epithelium and separates it from deeper tissues. Recent studies suggest that nanoscale topography of the surface of basement membrane may modulate adhesion, migration, proliferation and differentiation of overlying epithelium. This study was performed to elucidate nanoscale topographic features of basement membrane of the bladder. Bladder tissues were obtained from three adult female rhesus macaques. A process was developed to remove the epithelium while preserving the underlying basement membrane, and tissues were evaluated by immunohistochemistry and scanning electron microscopy (SEM). Detailed measurements were made of stereo SEM images to quantitatively define topographic features. Measurements made from multiple SEM images of bladder basement membrane provided the following values for topographic features: mean feature height, 178+/-57 nm; mean fiber diameters, 52+/-28 nm; mean pore diameter, 82+/-49 nm; and mean interpore distance (center to center), 127+/-54 nm. These dimensions are similar to those reported previously for basement membranes of other species and anatomical locations. This information provides a rational basis for design of nanostructured biomaterials to produce composite grafts for repair or replacement of segments of the urinary tract.

Animals↗
PubMed2003

Epithelial contact guidance on well-defined micro- and nanostructured substrates.

The human corneal basement membrane has a rich felt-like surface topography with feature dimensions between 20 nm and 200 nm. On the basis of these findings, we designed lithographically defined substrates to investigate whether nanotopography is a relevant stimulus for human corneal epithelial cells. We found that cells elongated and aligned along patterns of grooves and ridges with feature dimensions as small as 70 nm, whereas on smooth substrates, cells were mostly round. The percentage of aligned cells was constant on substrate tomographies with lateral dimensions ranging from the nano- to the micronscale, and increased with groove depth. The presence of serum in the culture medium resulted in a larger percentage of cells aligning along the topographic patterns than when no serum was added to the basal medium. When present, actin microfilaments and focal adhesions were aligned along the substrate topographies. The width of the focal adhesions was determined by the width of the ridges in the underlying substrate. This work documents that biologic length-scale topographic features that model features encountered in the native basement membrane can profoundly affect epithelial cell behavior.

Actin Cytoskeleton↗
PubMed2003

Non-enzymatic glycation in corneas from normal and diabetic donors and its effects on epithelial cell attachment in vitro.

BACKGROUND: Diabetic keratopathy manifests as persistent and recurrent erosions and delayed wound healing. To investigate the role of non-enzymatic glycation of proteins in the pathogenesis of diabetic keratopathy, we studied the presence of advanced glycation end products in corneas normal and diabetic donors, and the ability of human comeal epithelial cells to attach to non-enzymatically glycated extracellular matrix proteins. METHODS: Corneas from normal donors and donors with diabetes were sectioned and immunostaining was performed using a primary antibody that detects N(epsilon)-(carboxymethyl) lysine. Collagen I and fibronectin-coated tissue culture plates were non-enzymatically glycated by incubating with 500-mM glucose-6-phosphate for three weeks at 37 degrees C. To determine attachment corneal epithelial cells were allowed to adhere to glycated plates (1 to 4 hours, 37 degrees C). Non-adherent cells were removed by gentle washing and the number of attached cells was quantitated using Calcein-AM. RESULTS: Corneas from donors with diabetes showed more-intense immunostaining for N(epsilon)-(carboxymethyl) lysine than normal donors of similar ages. Non-enzymatic glycation of collagen I and fibronectin significantly reduced the ability of corneal epithelial cells to adhere to these extracellular matrix proteins. CONCLUSION: Our data suggest that non-enzymatically glycated proteins may interfere with attachment and hence migration of epithelial cells across the corneal surface, and therefore may contribute to some of the clinical manifestations of diabetic keratopathy. Diabetes leads to an accelerated aging process resulting in a cornea that may be much older than the chronological age of the patient. From a clinical perspective, particular care should be taken when considering the use of contact lenses and following corneal injury in these patients.

Adolescent↗
PubMed2002

Conjunctivitis in birds.

There are numerous causative agents of avian conjunctivitis, many of which are associated with systemic disease. A thorough evaluation of the patient's history and habitat is mandatory, and a complete physical examination and ophthalmic examination are required. The owner should be informed at the initial visit that an orderly sequential approach for obtaining diagnostic specimens is required and that a protracted course of treatment may be required to bring about complete, long-lasting resolution of clinical signs.

Animals↗
PubMed2002

Successful six-day kidney preservation using trophic factor supplemented media and simple cold storage.

This study examined the effect of trophic factor supplementation [TFS; bovine neutrophil peptide-1 (bactenecin), 1 mg/L; substance P, 2.5 mg/L; nerve growth factor, 20 microg/L; epidermal growth factor, 10 microg/L; insulin-like growth factor-1, 10 microg/L] during cold storage with UW lactobionate solution. Dogs transplanted with kidneys stored for 4days in TFS-UW had significantly lower peak serum creatinine values (mean 2.9 +/- 0.2mg/dL) and returned to normal values faster (6 days) than kidneys stored for 3days in unmodified UW solution (4.2 +/- 0.3 mg/dL and 14 days, respectively). Kidneys stored for 5days in TFS-UW (mean peak creatinine 3.7 +/- 0.3) functioned equivalently to kidneys stored for 3days and better than kidneys stored for 4 days in UW alone. Dogs with kidneys stored for 6days in TFS-UW had mean peak creatinines of 5.7 +/- 0.4 mg/dL. These returned to normal creatinine values in 14 days, equal to 3-day stored and significantly better than kidneys stored for 4days in UW alone (20days recovery time). This study shows trophic factor deprivation appears to be a critical mechanism of injury in organ preservation with current synthetic storage media, and marks the initial development of a synthetic biologically active preservation solution, the next generation of preservation media.

Adenosine↗
PubMed2002

Electron microscopy of the canine corneal basement membranes.

The purpose of this study was to characterize the surface topographical features of the epithelial and endothelial (Descemet's) basement membranes of the canine cornea. Corneas were obtained from young, healthy dogs (<2 years old) with no history or evidence of previous ocular disease. The epithelium and endothelium was carefully removed preserving the anterior and posterior basement membranes. The specimens were examined by transmission electron microscopy and scanning electron microscopy. The epithelial and endothelial basement membrane surface topography is an intricate meshwork of pores and fibers measuring in the nanometer size range. The features of the endothelial basement membrane overall are smaller in size than the epithelial basement membrane. These surface topographical features may incite changes in epithelial and endothelial cell behavior.

Animals↗
PubMed2002

The effect of chronic corneal epithelial debridement on epithelial and stromal morphology in dogs.

PURPOSE: To determine the effect of chronic corneal epithelial debridement on epithelial and stromal morphology and extracellular matrix components, and to compare those changes to those in spontaneous chronic corneal epithelial defects (SCCED) in dogs. METHODS: Axial corneal epithelial wounds, 10 mm in diameter, were created weekly for 8 weeks in five normal adult laboratory beagles. Slit lamp biomicroscopy and corneal pachymetry were performed weekly before wounding. Three days after the last debridement the dogs were killed humanely, and corneas were processed for light and electron microscopy and immunohistochemistry for collagen IV, collagen VII, fibronectin, and laminin. RESULTS: No significant changes in corneal thickness were found. All samples demonstrated epithelial dysmaturation adjacent to the wound edge, and, in four of five, a narrow zone of nonadherent epithelium formed adjacent to the exposed stroma. All samples had a stromal acellular zone in the area of the defect and continuing for a short distance under the adjacent attached epithelium. Experimentally wounded dogs did not form the superficial hyaline acellular lamina found in 92% of dogs with SCCED. Laminin, collagen IV, and fibronectin were present on the stromal surface in all samples, and collagen VII was present in four of five samples. Transmission electron microscopy (TEM) demonstrated the presence of basement membrane on the surface of the exposed stroma. CONCLUSIONS: Epithelial changes are similar between experimentally wounded dogs and dogs with SCCED. The stromal acellular zone that forms in experimentally wounded dogs is distinct from the hyaline lamina observed in dogs with SCCED. The difference in the acellular stromal layers between chronically wounded dogs and dogs with SCCED may be of relevance to our understanding of the pathophysiology of persistent epithelial defects.

Animals↗