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Biomedical subjects

J V Jester

Publications and source records attributed to J V Jester.

At least 127 records · Page 7Linked to original sources

Epithelial inclusion cysts following radial keratotomy.

A 26-year old woman underwent radial keratotomy combined with concentric trephination (6.5 and 8 mm) six months prior to development of multiple epithelial inclusion cysts. A biopsy specimen from one enlarging cyst showed an absence of scarring in addition to the retention of epithelium within a radial incision wound. Adjacent keratocytes were abnormal and seemed to be undergoing degenerative changes. To our knowledge, this is the first report of a failure in corneal wound healing associated with microcystic degeneration of the epithelium following radial keratotomy.

Adult↗

Analysis of incision depth following experimental radial keratotomy.

Eight-incision radial keratotomy was performed on a series of cadaver eyes using ultrasonic pachymetry and both metal and diamond blades. There were no statistically significant differences in mean corneal flattening (approximately 9.5 diopters) obtained with either the diamond or metal blades. Short diamond knife incisions produced less flattening that was statistically significant (5.8 diopters). Histologic analysis of incision depth revealed similar results with both the diamond (84%) and metal (86%) blades.

Cadaver↗

Hematoporphyrin derivative photoradiation therapy for the treatment of intraocular tumors: examination of acute normal ocular tissue toxicity.

Preclinical studies designed to define potential side effects resulting from the use of hematoporphyrin derivative (HPD) photoradiation therapy (PRT) as a modality for treating intraocular tumors have been performed. Pigmented rabbits were used to evaluate acute normal ocular tissue toxicity following single HPD PRT treatments in which the light was directed through the pupil and onto a 1-sq cm area of the retina. The treatment procedure consisted of the i.v. administration of HPD (1 to 10 mg/kg) followed 48 hr later by a 15-min exposure of localized red light [635 +/- 5 nm; 40 to 400 milliwatts/sq cm] generated by a free running rhodamine B dye laser pumped by a 5-watt argon laser. Toxicity to normal retinal tissue was documented using fundus photography, fluorescein angiography, and histological examination. The results of this study demonstrated that ocular damage in the form of retinal edema, detachment, and necrosis could be induced by clinically relevant doses of HPD PRT. The area of retinal damage was limited to the treatment field in all but the highest doses of HPD PRT. The histological results were in agreement with the visual observations in that abrupt and demarcated transition areas between injured and normal-appearing retina were observed. Care will have to be used in the delivery of light to the treatment field if HPD PRT is to be utilized for treatment of intraocular tumors.

Animals↗

The effect of radial keratotomy on ocular integrity in an animal model.

The safety of deep corneal incisions in radial keratotomy was evaluated in a porcine model of blunt trauma. One eye of each enucleated pair (right and left) of porcine eyes was subjected to a variation of radial keratotomy; the fellow eyes served as unoperated-on controls. All eyes were subjected to a standard injury. Control eyes ruptured at the equatorial sclera. Eyes with radial incisions cut through approximately 70% of corneal thickness also ruptured at the equator. When incisions of this depth (70%) were extended across the limbus (rather than to the corneal-scleral junction), all ruptures occurred at the limbal incisions. Eyes cut 95% to 100% of corneal thickness tended to rupture at the incisions. The safety of deep radial keratotomy incisions with respect to ocular integrity is discussed.

Animals↗

Type I collagen and fibronectin synthesis by retrocorneal fibrous membrane.

The primary cultures obtained from the experimentally induced retrocorneal fibrous membrane synthesized and secreted into the medium mainly type I procollagen. This collagen was characterized after limited pepsin treatment and identified as type I collagen by the following criteria: (1) it contained two alpha 1 chains and one alpha 2 chain, (2) its sedimentation behavior was identical to that of type I collagen from skin, and (3) its peptide map after limited proteolysis with Staphylococcus aureus V8 protease was identical to that of type I collagen. The medium contained procollagen I, which was converted into alpha size chains by limited pepsin treatment, whereas the cellular fraction contained type I collagen already processed to its end product. Type III collagen and basement membrane collagen were present as minor components in this system Fibronectin, one of the major glycoproteins in extracellular matrices, was also synthesized and secreted into the medium. In contrast, normal corneal endothelial cells produce mainly basement membrane collagen.

Animals↗

In vivo biomicroscopy and photography of meibomian glands in a rabbit model of meibomian gland dysfunction.

The usefulness of transillumination and biomicroscopy of the lid margin in assessing meibomian gland dysfunction was studied in a rabbit model. Transillumination of rabbit lids treated with topical epinephrine for 2 to 3 months revealed plugging of the meibomian gland orifice. Plugging appeared to be correlated histopathologically with increased thickness and hyperkeratinization of the ductal epithelium at the orifice. Continued treatment resulted in microcystic changes within the duct, which were not easily discernible by routine examination without the aid of a transilluminator. Cystic changes were correlated with dilation of the duct by retained desquamated cornified cells. Biomicroscopy proved valuable in identifying and documenting early lesions associated with epinephrine-induced meibomian gland dysfunction.

Animals↗

Radial keratotomy in non-human primate eyes.

We performed a prospective study of the effects of radial keratotomy in the owl monkey. We compared a 16-incision and an eight-incision radial keratotomy, and followed the changes in corneal curvature, corneal thickness, endothelial cell counts, and intraocular pressure. We compared the results of the changes in these clinical factors with a histopathologic and ultrastructural analysis of the time-related changes after radial keratotomy in another group of animals. We found that the loss of initial corneal flattening following radial keratotomy corresponded with the contracture of the wound as demonstrated by histopathologic and ultrastructural study. This procedure results in a significant endothelial cell loss (14% to 15%), which is a result of the postsurgical inflammation associated with this surgery. Additionally, examination of the histopathologic structure of these corneas showed a high level of variability in the surgical incision depth, which we believe is responsible for the marked variations in the response to the surgical procedure.

Animals↗

A statistical analysis of radial keratotomy in human cadaver eyes.

We performed radial keratotomy using the Fyodorov techniques on 14 Eye Bank eyes. Using stepwise regression and best subset statistical analysis, we examined the effects of incision length. We found that the most significant variables affecting postoperative corneal curvature were the incision depth and length. The histologic examination of serial sections of the Eye Banks eyes after this procedure revealed that the average achieved incision depth, using a standard blade depth setting, had variations of up to 30% between eyes. This inability to standardize incision depth may lead to the variable surgical results reported in radial keratotomy.

Cadaver↗

Radial keratotomy in fresh human cadaver eyes.

Radial keratotomy in fresh human cadaver eyes produced corneal flattening varying from 6 to 11 diopters (D). There was no significant difference in the effectiveness of the incisions to the incisions to the limbus compared to incisions through the limbus. Eighty to ninety percent of the flattening effect was obtained after the first eight incisions. The preoperative keratotomy reading was not helpful in predicting the final result. Histopathology of the incised corneas revealed considerable variation in incision depth and demonstrates the difficulty in achieving deep incisions safely.

Aged↗

Preliminary study of keratomileusis in primates (Macaca speciosa).

Keratomileusis was investigated in five stumptail monkey eyes (Macaca speciosa). An arbitrary flattening of six diopters was selected and programmed in the computer for each eye. The actual flattening achieved two weeks after surgery ranged from -4.00 D to -6.00 diopters (D). One to 5 diopters of the initial effect was lost during the six-month observation period. Histopathology of two eyes demonstrated attenuation and/or loss of Bowman's membrane with overlying epithelial defects.

Animals↗

Modification of corneal curvature following radial keratotomy in primates.

Radial keratotomy was performed on six stumptail monkey eyes (Macaca speciosa) and 22 owl monkey eyes (Aotus trivirgotus). Changes in keratometry, specular microscopy, and tonometry have been studied postoperatively for three to six months. Sixteen radial incisions in stumptail monkey eyes resulted in a significant mean corneal flattening of 2.75 diopters (P less than 0.005). This effect was stable with a mean 2.50 diopters flattening remaining six months after surgery. Sixteen radial incisions in ten owl monkey eyes resulted in a much greater short-term effect, with a mean 10.50 diopters of flattening present at two weeks postoperatively. This flattening deteriorated over the observation period, and no significant effect was noted three months after surgery. A comparison of 8 vs 16 incisions on owl monkey eyes demonstrated that these two procedures are equally effective in initially flattening the cornea. Complications and side effects encountered included perforations, irregular astigmatism, corneal neovascularization, transitory increase in pachometry, and decrease in central corneal endothelial cell density in isolated cases.

Animals↗

Meibomian gland studies: histologic and ultrastructural investigations.

Heightened interest in meibomian gland dysfunction has prompted us to evaluate the normal morphological and ultrastructural characteristics of the meibomian gland. Histologic analysis of human, primate, steer, and rabbit glands revealed evidence of keratinized epithelium extending throughout the meibomian gland duct. Characteristic ultrastructural features of keratinized epithelium identified in primate and rabbit glands included tonofilaments, keratohyaline granules, lamellar bodies, and keratinized squamous cells. Comparison of the meibomian gland duct to the pilosebaceous canal and the sebaceous duct brought out certain dissimilarities such as (1) the lack of a well-developed stratum granulosum and (2) the absence of lipid inclusions within transitional cells from duct to acini. We postulate that abnormalities of the keratinizing process may be responsible for meibomian gland dysfunction states.

Animals↗

Pathology of ocular irritation with bleaching agents in the rabbit low-volume eye test.

Despite differences in the processes leading to tissue damage, the ocular irritation response to various surfactants, two concentrations of an acid and an alkali, and an acetone, alcohol, aromatic amine, and aldehyde has been shown to depend on the extent of initial injury. The purpose of this study was to assess the extent to which this fundamental relationship exists for bleaching agents in the rabbit low-volume eye test. Ten microl of sodium perborate monohydrate (NaBO3), sodium hypochlorite (NaOCl), 10% hydrogen peroxide (H2O2), and 15% H2O2 was applied directly to the cornea of the right eye of each rabbit. Macroscopic assessments for irritation were made 3 hours after dosing and periodically until 35 days. Light microscopic examinations were conducted on tissues obtained at 3 hr and on 1, 3, and 35 days. In vivo confocal microscopy (CM) and measurements of dead corneal epithelial cells and keratocytes at 3 hours and 1 day were used to characterize quantitatively initial corneal injury, while in vivo CM performed at 3 hours and 1, 3, 7, 14, and 35 days was used to characterize quantitatively the corneal changes over time. The changes with NaBO3 and NaOCl were consistent with mild irritancy. For both, corneal injury was limited to the epithelium and superficial stroma. The changes with 10% H202 and 15% H2O2 were consistent with severe irritation. Both concentrations affected the epithelium and deep stroma, with 15% H2O2 also at times affecting the endothelium. However, unlike other irritants previously studied, with 10% H2O2 and 15% H2O2 there was an incongruity between the extent of epithelial and stromal injury, with stromal injury being more extensive than epithelial injury. A similar, although less dramatic, effect was observed with NaBO3. Additionally, there was still significant keratocyte loss at 35 days with 10% H2O2 and 15% H2O2 even though the eyes at times were considered to be macroscopically normal. These observations highlight the need to include both epithelial and stromal components in an ex vivo or in vitro alternative assay. In conclusion, these results continue to support and extend our hypothesis that ocular irritation is principally defined by the extent of initial injury despite clear differences in the means by which irritants cause tissue damage. Importantly, we have identified unique differences in the ocular injury and responses occurring with bleaching agents that are important to consider in the development and validation of alternative ocular irritation tests to characterize a broad range of materials differing in type and irritancy.

Animals↗

Pathology of ocular irritation with acetone, cyclohexanol, parafluoroaniline, and formaldehyde in the rabbit low-volume eye test.

The ocular irritation responses to 11 different surfactants and two concentrations of acetic acid and sodium hydroxide have been shown to depend on the extent of initial injury, despite marked differences in the processes leading to tissue damage. The purpose of these studies was to determine the extent to which this fundamental relationship applies to other nonsurfactants. Ten microl of acetone (ACT). cyclohexanol (CY), parafluoroaniline (PF), or 37% formaldehyde (FA) was directly applied to the cornea of the right eye of each rabbit. Eyes and eyelids were macroscopically scored for signs of irritation beginning 3 hours after dosing and periodically until recovery or 35 days. Tissues were obtained for light microscopic examination after 3 hours and on days 1, 3, and 35. Initial corneal injury was characterized quantitatively at 3 hours and I day using in vivo confocal microscopy (CM) and by postmortem quantitation of dead corneal epithelial cells and keratocytes using a Live Dead Assay (L/D, Molecular Probes) and scanning laser CM. Corneal changes over time were characterized quantitatively using in vivo CM performed at 3 hours and 1, 3, 7, 14, and 35 days. The changes with ACT were consistent with mild irritation. Corneal injury was limited to the epithelium and superficial stroma, with the mean normalized depth of injury (NDI) being less than 10% with the majority of regions showing no stromal injury. Changes with CY and PF were consistent with moderate to severe irritation, and FA caused severe irritation. Specifically, corneal injury by CY and PF tended to involve the epithelium and anterior stroma, with the mean NDI being 10.4% to 23.8%, while injury with FA involved the epithelium, deep stroma, and at times the endothelium. Interestingly, with FA significantly less injury was observed at 3 hours with a dramatic increase in injury observed at 1 day and thereafter. In conclusion, these results continue to support and extend our hypothesis that ocular irritation is principally defined by the extent of initial injury despite clear differences in the means by which irritants cause tissue damage. We believe this approach can be applied to developing alternative assays based on injury to ex vivo eyes or injury to an in vitro corneal equivalent system.

Acetone↗

Effect of cell migration on the maintenance of tension on a collagen matrix.

Although it is known that cells promote structural reorganization of the collagen architecture, how individual cells exert mechanical tension on the matrix is not clearly understood. In the present study we have investigated the mechanical interaction of individual corneal fibroblasts with a collagen matrix using an improved version of our previously described in vitro force-measurement system (Roy, P. et al. Exp. Cell Res. 232:106-117, 1997). The elastic distortion of the collagen matrix exerted by cells was temporally recorded and analyzed using a two-dimensional finite-element model to quantify the forces exerted on the matrix. Time-lapse videomicroscopy of serum-cultured cells on the matrix for up to 6 h revealed that individual fibroblasts generated measurable tension on the matrix during pseudopodial extension and slow retraction. Fast retraction, an event observed during active cell migration, was associated with dramatic release of tension on the matrix. An apparent inverse correlation was observed between cell translocation and maintenance of matrix tension. Additional experiments with cells under serum-free conditions revealed that these cells fail to generate any detectable tension on the matrix despite undergoing filopodial extension and retraction. Since serum-free cells do not form focal adhesions or stress fibers, these experimental data suggest that contractility of nonmotile cells, coupled with strong cell-matrix adhesion, is the most favorable mechanism of generating and maintaining tension on the extracellular matrix.

Adhesiveness↗

Quantitative characterization of acid- and alkali-induced corneal injury in the low-volume eye test.

Defining the extent of initial injury has proven to be a useful basis for differentiating the ocular irritation potential of surfactants; however, the applicability of this method to other types of irritants has not been demonstrated. In the following studies we characterized the extent of corneal injury following exposure to different concentrations of acetic acid and sodium hydroxide (NaOH) in the rabbit low-volume eye test. Groups of rabbits received 3% acetic acid, 10% acetic acid, 2% NaOH, or 8% NaOH and were evaluated in vivo by macroscopic and in vivo confocal microscopic examination and postmortem using a live/dead staining kit and scanning laser confocal microscopic examination. Quantitative assessment of macroscopic scores, corneal surface epithelial cell size, corneal epithelial thickness, corneal thickness, depth of stromal injury, corneal light scattering (confocal microscopy through focusing, CMTF), and number of dead cells was conducted at various times, including the following: at 3 hours and at 1, 3, 7, 14, and 35 days. Based on macroscopic scores, the order of ocular irritancy potential was 3% acetic acid < 2% NaOH < 10% acetic acid < 8% NaOH. Evaluation of the quantitative in vivo and postmortem microscopic live/dead data revealed a slight decrease in epithelial thickness and an increase in dead epithelial cell numbers with 3% acetic acid. With 2% NaOH, significant focal changes in epithelial cell size, epithelial thickness, corneal thickness, and number of dead surface epithelial cells occurred at 3 hours and at 1 day, with injury to only a very small number of corneal stromal keratocytes, despite the presence of epithelial denudation. Changes with 10% acetic acid were similar to those noted with 2% NaOH at 3 hours and 1 day, but these changes were more diffuse and included stromal injury to a depth of 7.2 +/- 9.3% of the corneal thickness, with significant numbers of dead keratocytes. Eight percent NaOH, on the other hand, caused focally extensive injury that averaged 26.3 +/- 18.4% of the corneal thickness at 1 day, with significant light scattering from the cornea, which did not return to normal by 35 days postinjury. Overall, these data indicate that ocular irritation as a result of acetic acid and NaOH was associated with changes similar to those observed with surfactants (ie, slight irritants damage the corneal epithelium, mild and moderate irritants damage the corneal epithelium and anterior stromal cells, and severe irritants damage the corneal epithelium and deep stroma). To our knowledge, this is the first time that the ocular irritation potential for different types of materials (acid/alkali, surfactants) has been shown to be primarily dependent on the initial area and depth of injury.

Acetic Acid↗