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H D Cavanagh

Publications and source records attributed to H D Cavanagh.

At least 19 recordsLinked to original sources

In vivo confocal microscopy in clinical dental research: an initial appraisal.

Until recently, the in vivo microscopic investigation of intraoral tissues at high resolution has been virtually impossible. Confocal microscopy enables high-resolution imaging to be achieved below semitransparent surfaces in intact living specimens, but this may still be impractical for intraoral applications because of the need to stabilize the sample. The development of a steadying objective (x 240 overall mag.) which is held against the sample surface and is focused by moving internal elements, avoids the need for fine adjustment of the living sample under the microscope to achieve a change of focus. It is therefore more comfortable and also reduces the problems of movement due to the pulse. The objective was used with a tandem scanning microscope, with images recorded via a SIT video camera. Using this system internal tooth structure (e.g. enamel prisms/adhesive restoration interfaces) and the lining cells of the gingival crevice through to the junctional epithelium may be examined. It is also possible to image the oral mucous membrane, focusing to the capillary loops in the basal layers, where streaming red blood cells can be seen. Access is limited to the anterior regions as far back as the premolar teeth. Applications could include caries research, soft and hard tissue responses to biomaterials (e.g. implants), wound healing and monitoring the effect of periodontal treatment regimens. This new technique offers numerous exciting opportunities for the microscopic investigation of many clinical operative procedures in vivo, allowing the response of the tissues to be non-destructively monitored, over time, at high resolution.

Composite Resins

Effect of contact-lens-induced hypoxia on lactate dehydrogenase activity and isozyme in rabbit cornea.

Lactate dehydrogenase (LDH) levels and subunit isozyme patterns in cornea were monitored in 36 albino rabbits wearing thick, rigid, gas-permeable contact lenses for periods of 24 h, 2 and 7 days, and 1 and 3 months. The oxygen transmissibility of the contact lens was 15.3 x 10(-9) (cm/s) (ml O2/ml mm Hg). The activity of LDH in corneal tissue decreased according to the duration of lens wear. The LDH isozyme patterns shifted with lens wear from LDH1,2,3 (heart type, aerobic) to LDH4,5 (skeletal muscle type, anaerobic). The cornea swelled 8.8% with overnight contact lens wear, with increased swelling (11-12%) after further continuous lens wear. After contact lens removal, however, the LDH activity and the isozyme pattern returned to normal, and the corneal thickness quickly returned to normal. Based on these observations, it is suggested that LDH in rabbit corneas was physiologically affected by lens-induced hypoxia, but these changes were reversible. These results might further suggest that tear LDH levels in the human contact lens wearer could provide an ongoing assessment of the tolerance of the lens by the ocular surface.

Animals

Effects of increasing Dk with rigid contact lens extended wear on rabbit corneal epithelium using confocal microscopy.

The effects of 24-h wear of various Dk-rigid gas-permeable (RGP) contact lenses on the rabbit corneal epithelium were studied by in vivo tandem scanning confocal microscopy (TSCM), and confirmed by scanning electron microscopy (SEM). Lenses used were polymethylmethacrylate (PMMA) (Dk/L = 0), RGP experimental A lens (siloxanylmethacrylate-fluoromethacrylate-methylmethacrylate , 33), experimental B (siloxanylmethacrylate-fluoromethacrylate, 56), and experimental C (siloxanylstyrene-fluoromethacrylate copolymer, 64 x 10(-9)) (cm/s) (ml O2/ml mm Hg) with 0.15-mm thickness (Dk/L measured by polarograph including boundary layer effect). After 24-h PMMA lens wear, TSCM showed no superficial epithelial cells but only exposed, underlying wing cells. The cornea with experimental A showed partial superficial epithelial desquamation. With experimental B wear, slight superficial epithelial cell swelling and desquamation were observed on the surface of the cornea. No changes were observed for the eye with experimental C and control. The observed severity of desquamation of superficial epithelial cells was dependent on the oxygen transmissibility (Dk/L) of RGP lenses worn. All in vivo findings were confirmed by SEM observations. Based on the results of this study, we conclude that (a) although Dk/L = 56 lens B shows no residual overnight corneal swelling, surface damage is still produced; (b) Dk/L = 64 lens C is best for epithelium showing the same corneal images as control; and (c) TSCM is a good way to evaluate the contact lens safety and efficacy in vivo at the cellular level noninvasively.

Animals

Digital image acquisition in in vivo confocal microscopy.

A flexible system for the real-time acquisition of in vivo images has been developed. Images are generated using a tandem scanning confocal microscope interfaced to a low-light-level camera. The video signal from the camera is digitized and stored using a Gould image processing system with a real-time digital disk (RTDD). The RTDD can store up to 3200 512 x 512 pixel images at video rates (30 images s-1). Images can be input directly from the camera during the study, or off-line from a Super VHS video recorder. Once a segment of experimental interest is digitized onto the RTDD, the user can interactively step through the images, average stable sequences, and identify candidates for further processing and analysis. Examples of how this system can be used to study the physiology of various organ systems in vivo are presented.

Animals

Comparison of in vivo and ex vivo cellular structure in rabbit eyes detected by tandem scanning microscopy.

Using the tandem scanning microscope, in vivo confocal microscopic images of living eyes were compared to images obtained from ex vivo, freshly enucleated or fixed tissue in the rabbit. In the normal cornea, microscopic details of the superficial epithelium, basal lamina, stromal fibrocyte nuclei, nerves and endothelial cell borders were easily discernible. Removal of the eye from the intact animal resulted in loss of detail with distortion of the normal structural interrelationships within the corneal stroma whilst enhancing details of the corneal epithelium. Formalin fixation further enhanced details of the basal and suprabasal corneal epithelial cell nuclei and the stromal fibrocyte cell borders whilst inducing prominent brightly reflecting folds in the thickened stroma with concomitant enhancement of the edge contrast of the collagen lamellae. These changes appeared to be related, in part, to hydration of the cornea and artefactual pooling of water between structures that may enhance reflectivity by increasing the difference between the refractive index of the cellular and extracellular elements. We conclude that microscopic examination of ex vivo preparations of corneal tissue, although providing increased resolution similar to conventional light microscopic techniques, significantly altered the normal structural relationships and could lead to erroneous measurements of the physiological properties of the tissue as compared to in vivo microscopy of undisturbed, intact tissue.

Animals

A quantitative method for LDH, MDH and albumin levels in tears with ocular surface toxicity scored by Draize criteria in rabbit eyes.

We have established a quantitative method for evaluation of ocular surface lesions on the basis of lactate dehydrogenase (LDH) activity, malate dehydrogenase (MDH) activity, and albumin levels in rabbit tears. Lesions were produced with solutions of 0.005-0.02% benzalkonium chloride (BAK), 0.01-0.03% chlorhexidine digluconate (CHX), and 0.01-0.03% polyhexamethylene biguanide hydrochloride (PHMB), all of which are widely used in topical ophthalmic preparations. Two drops of test solution were instilled 15 times into rabbit eyes at 5 minute intervals. Sixty minutes after 0.02% BAK instillation, tear LDH activity increased from 1,840 U/L (without instillation) to 26,100 U/L, and concomitantly tear albumin levels rose from 0.11 mg/mL (without instillation) to 9.48 mg/mL. Instillation of 0.03% CHX and 0.03% PHMB caused smaller increases in LDH and MDH activity and albumin tear levels. LDH activity and albumin levels in tears were significantly correlated with the degree of total ocular surface lesions in both cornea and conjunctiva as observed by slit lamp biomicroscopy quantified using a modified Draize score. Based on the results of this study, we believe that LDH activity and albumin level in tears can be used as objective indicators for the quantitative evaluation of ocular surface lesions on both cornea and conjunctiva following application of topical ophthalmic preparations containing cytotoxic preservatives in animals and man.

Albumins

Radial keratotomy. 1. The wound healing process and measurement of incisional gape in two animal models using in vivo confocal microscopy.

Using in vivo confocal microscopy, corneal wound healing was evaluated in both rabbit and cat eyes after radial keratotomy. A total of six rabbit and six cat eyes were evaluated sequentially over time for 1 mo after surgery by in vivo confocal microscopy, and quantitative measurements of changes in incisional wound gape were determined. In vivo histopathologic changes were correlated with conventional histopathologic findings in 18 rabbit and 4 cat eyes; the animals were killed at various intervals from 0-30 days after surgery. In the rabbit, in vivo corneal wound healing was characterized by the initial ingrowth of corneal epithelium followed by persistence within the wound without a marked fibrotic response. Measurement of incisional wound gape showed increasing gape from 144 +/- 32 microns on day 0 to 976 +/- 155 microns on day 26 at a distance of 2.4 mm from the optical zone. These in vivo measurements were not significantly different (P = 0.996) from those obtained using conventional histopathologic techniques which showed an incisional wound gape of 252 +/- 112 microns on day 0 and 917 +/- 216 microns on day 26 at 2.5 mm from the optical zone. In the cat eyes, healing of radial keratotomy wounds showed an initial increase in incisional wound gape from 135 +/- 56 microns on day 0 to 245 +/- 88 microns on day 7 at a distance of 2.4 mm from the optical zone. Starting at day 14 and continuing to day 30, there was a progressive decrease in incisional wound gape from 198 +/- 41 microns to 92 +/- 35 microns. Sequential, in vivo histopathologic analyses indicated that increasing incisional wound gape correlated with the retention of corneal epithelium in the wound. Initiation of decreasing incisional wound gape was associated with replacement of the incisional epithelial plug with fibroblastic tissue. These changes in the incisional wound gape observed in the cat suggest that healing of radial keratotomy wounds involves contraction of the wound in response to the ingrowth of fibroblastic cells. Furthermore, the contractile response appears to be biphasic involving a precontractile and contractile phase. Overall these data indicated that in vivo confocal microscopy provides quantitative histopathologic data on living tissue comparable with that obtained with conventional techniques on dead, fixed, and sectioned tissue. Additionally, the absence of wound fibrosis in the rabbit radial keratotomy model raises important questions as to the appropriateness of this experimental model for human radial keratotomy.

Animals

Radial keratotomy. II. Role of the myofibroblast in corneal wound contraction.

The cellular mechanism of corneal wound contraction after radial keratotomy (RK) was studied in a feline eye model. A total of 10 cat eyes were evaluated at various times from 0-30 days after surgery. Changes in the distribution of intracellular filamentous actin, nonmuscle myosin, alpha-actinin, surface membrane alpha 5 beta 1 integrin, and extracellular fibronectin were studied using immunofluorescence and laser confocal and electron microscopy. From day 3-7, staining for fibronectin increased along the wound margin. By day 7, keratocytes adjacent to the wound margin showed increased f-actin staining with intense staining for fibronectin compared with normal keratocytes. Myosin and alpha 5 beta 1 integrin expression was very weak at this time; alpha-actinin was not found. By day 14, fibroblasts within the wound formed f-actin microfilament bundles (stress fibers) which colocalized with fibronectin. Wound-healing fibroblasts also stained positively for alpha 5 beta 1 integrin, myosin, and alpha-actinin (the latter two were colocalized). The presence of myosin and alpha-actinin in the wound fibroblasts and the re-organization of f-actin into stress fibers by day 14 correlated with the development of wound contraction. A comparison of the cellular distribution of actin, myosin, and alpha-actinin with alpha 5 beta 1 integrin 14 days after injury suggested that integrin was localized along stress fiber bundles during wound contraction. The data from this study suggest that modulation of wound gape during healing of RK wounds may involve transformation of the corneal keratocyte to a myofibroblast-like cell and the subsequent formation of intracellular stress fibers composed of f-actin, nonmuscle myosin, and alpha-actinin. Based on the colocalization of fibronectin filaments and f-actin filaments and the unique distribution of alpha 5 beta 1 integrin, these findings support the hypothesis that the tension within the wound is generated by the formation of intracellular stress fibers and the interactions between stress fibers and the extracellular matrix, mediated by specific membrane receptor molecules.

Animals

Radial keratotomy. III. Relationship between wound gape and corneal curvature in primate eyes.

The relationship between changes in wound gape and corneal curvature after radial keratotomy (RK) was evaluated in five primates. Four-incision RK was performed using a diamond knife set to 100% of central corneal thickness with a 3-mm optical zone. In vivo measurements of wound gape were obtained using tandem scanning confocal microscopy at 3, 7, 14, and 45 days after surgery. The changes in corneal contour were measured at the same time points using a corneal modeling system with a specially designed primate cone. Wounds progressively increased in width to a maximum of 38 +/- 1 microns (n = 5) at day 7. After day 7, wounds showed increasing fibrosis which correlated with decreasing wound gape to 20 +/- 1 microns at day 45. A similar temporal change was detected in central corneal curvature (K), with maximum flattening occurring at day 7 (delta K = -3.17 +/- 0.90 diopters, n = 5), and progressive regression of effect to -1.32 +/- 0.61 diopters (n = 5) at day 45. Although there was interanimal variation, the mean temporal changes in corneal curvature significantly paralleled the changes in wound gape (r = -0.96, n = 4, P < 0.05). Based upon these findings, a simple geometric model was proposed which provides a hypothetic foundation for the relationship between corneal curvature and wound gape after RK. Calculations of wound gape made from this analytic model (using the measured topographic data) showed significant correlation with the actual wound gape measurements (r = 0.96, n = 4, P < 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)

Animals

A prospective multicenter clinical trial to assess safety and efficacy of Menicon SF-P RGP lenses for extended wear.

A prospective, multicenter, national clinical trial was undertaken to assess the safety and efficacy of Menicon SF-P (melafocon A) rigid gas permeable (RGP) contact lenses for extended wear in healthy human eyes. Ten investigators enrolled 167 patients (334 eyes), of which 143 (286 eyes) were initially fit with lenses for the 12-month study. Most subjects were reexamined at 18 months or more, with findings essentially unchanged from those noted at 1 year. Two hundred-two eyes (71%) completed the study. Seventy-two eyes (25%) discontinued, and 12 (4%) elected to continue extended wear. During the study, no significant adverse clinical reaction attributable to the lens was observed. Surface irritation, occasional punctate staining, and lens binding were very rare. The average wearing time was 6.2 days, mandated partially by FDA restrictions as well as patient response. As expected, keratometric shifts were principally vertical: less than 0.99 D (69%), 1.00-1.99 D (28%), and 2.00-2.99 D (4%). The majority of changes observed were toward sphericity. Two hundred lens replacements were required, 24% for adjustments in power/base curve. Only 16 lenses were replaced for warpage, and none for discoloration or crazing. Of those completing the study, 76% rated overall lens comfort and satisfaction to be very good. These results suggest that RGP lens extended wear can be safe and effective and offer patients an acceptable alternative to extended wear of disposable hydrogel lenses.

Adolescent

Ciliated respiratory-like epithelium forming cystic conjunctival lesions in a patient with Stevens-Johnson syndrome.

A woman with severe cicatricial conjunctival changes secondary to allopurinol-induced Stevens-Johnson syndrome developed bilateral palpebral conjunctival cysts. Pathologic examination of these lesions revealed respiratory-like pseudostratified ciliated epithelium in the walls of several cysts of one eye. To our knowledge, this is the first report of this phenomenon.

Aged

Tear lactate dehydrogenase levels. A new method to assess effects of contact lens wear in man.

Lactate dehydrogenase (LDH) activity in tears was measured in 202 myopic rigid gas permeable (RGP) and 79 hydrogel human contact lens wearers and 48 normal controls by noninvasive microcapillary sampling. The oxygen permeabilities (Dk) of five selected RGP contact lenses ranged between 0 and 230 x 10(-11) (cm2/s) ml O2/ml mm Hg), and the water content (WC) of the hydrogel lenses was 38, 72, and 80%. When normal diurnal variation of tear LDH activity and tear sample volume (0.3-0.6 microliters) were carefully controlled, the tear LDH activity of RGP lens wearers in daily and extended wear correlated as an inverse function of Dk/L, with the highest enzyme activity observed in wearers of polymethylmethacrylate daily wear lenses (347.4 U/L). The tear LDH activity in Menicon EX (Dk 108) lens wearers was higher in the extended wear (192.7 U/L) than in the daily wear group (161.9 U/L). There was no significant difference in tear LDH activity between the Menicon SF-P (Dk 230) extended wear group (132.0 U/L) and controls (133.5 U/L). In the hydrogel lens daily wear group, tear LDH activity of Experimental 72 (WC 72%) and Experimental 80 (WC 80%) was higher than that of hydroxyethylmethacrylate (HEMA) despite having high Dk value. Experimental 80 extended wearers showed lower LDH activity in tears sampled between 0.3 and 0.6 microliters than that of HEMA wearers. These results suggest that sequential measurement of LDH levels in tears may offer a new and unique method for the assessment of the physiologic effects of contact lens wear on the ocular surface, and provide a new clinical paradigm for the interaction of the contact lens with the cornea.

Adolescent

Variations in corneal wound healing after radial keratotomy: possible insights into mechanisms of clinical complications and refractive effects.

Ultrastructural and histopathologic analysis was performed on three human corneal specimens for variable and complicated refractive outcomes 1-2 years after radial keratotomy. Specimens were obtained immediately postsurgery after microkeratome resection with homoplastic lamellar keratoplasty (two cases) and penetrating keratoplasty (one case) for correction of glare, severe astigmatism, overcorrection, and/or double vision. All three cases showed variability of wound healing and delayed corneal wound healing sites; epithelial retention cysts, and/or absence of stromal scar collagen that was not dependent on the length of time after surgery. Two of the three radial keratotomy specimens also contained extensive duplication of the superficial corneal epithelial basal lamina. When present, the thickened basal lamina (3-6 microns in thickness) was seen between all incisions evaluated and appeared to extend from the central optical zone out to the periphery of the lamellar button. The one full-thickness keratoplasty specimen showed focal loss of underlying endothelial cells with occasional migrating cells seen by scanning electron microscopy. These data support previous findings that delayed corneal wound healing with epithelial retention cysts remains the most common histopathologic alteration after radial keratotomy. The effects of variations and delay in wound healing between individuals could explain the lack of predictability of refractive outcome and continuing refractive instability in long-term follow-up after single or repeat radial keratotomy surgeries.

Adult

Confocal microscopic studies of living rabbit cornea treated with benzalkonium chloride.

The effects of benzalkonium chloride (BAK) on the living rabbit cornea were studied by in vivo Tandem scanning confocal microscopy (TSCM) and confirmed by conventional scanning electron microscopy (SEM). Two drops of saline or phosphate-buffered saline (PBS) containing BAK in concentrations of 0.02, 0.01, and 0.005% was applied to rabbit eyes 15 times at 5-min intervals. The solutions were pH 5.5-5.9 (saline) and pH 7.5 (PBS), and osmolarity was 275-280 (saline) and 300-307 mOsm (PBS). Immediately after application of 0.02 and 0.01% BAK, no normal corneal superficial epithelial cells could be imaged by in vivo TSCM. No swelling of the superficial epithelial cells was observed for the control solution without BAK; however, there was a small amount of desquamation. Application of as little as 0.005% BAK caused the superficial epithelial cells to swell and desquamate. The observed desquamation of corneal superficial epithelial cells increased with higher BAK concentrations applied to the eye. One hour after final drug application, inflammatory cells appeared on the surface of the cornea treated with 0.02% BAK. These findings were correlated with SEM observations. Based on the results of this study, we believe that BAK used frequently can produce clinical corneal toxicity and that the cytotoxicity of any topical ophthalmic solutions can be evaluated by in vivo TSCM.

Animals

Bilateral progressive essential iris atrophy and keratoconus with coincident features of posterior polymorphous dystrophy: a case report and proposed pathogenesis.

We report the first case known to us of an apparent bilateral association of essential iris atrophy (EIA) and keratoconus (KC), with coincident features of posterior polymorphous dystrophy (PPD). Based on this case and the published natural history and findings of both the irido corneal endothelial (ICE) syndrome and PPD, we propose a new hypothesis for the pathogenesis of the ICE syndrome with associated KC and/or PPD. We suggest that, similar to the genetics of retinoblastoma, the predisposition for either the ICE syndrome or for PPD is inherited as an inactive allele, the so-called "first hit." Inactivation of the second allele, or "second hit," which could occur at any time, might be the product of the background mutation rate or of an environmental trigger. Dedifferentiation or an abnormality in normal development could occur after the first or second hit, resulting in varying clinical patterns. We also concur with other investigators that PPD could be part of the spectrum of the ICE syndrome, owing to similarities in their clinical presentations, histopathology, specular and electron microscopy, and natural history.

Atrophy

Tandem scanning confocal microscopy (TSCM) of normal and ischemic living kidneys.

Tandem Scanning Confocal Microscopy (TSCM) allows one to section optically into and record real-time images of living organs and tissues in a noninvasive fashion. In this paper, we will present some initial TSCM observations of subcapsular nephrons in the living, intact kidneys of Munich-Wistar rats and evaluate the nephron's responses to temporary ischemia and to intravenous infusion of mannitol. The rats were anesthetized with Inactin and a laparotomy performed to expose the kidneys. Using a TSCM equipped with a 20 x water-immersion objective, we optically sectioned through the intact kidney capsule and recorded real-time images of living subcapsular glomeruli and uriniferous tubules. The proximal tubule brush border was highly reflective and allowed us to distinguish between the first and second segments of the proximal tubules as well as the distal tubules. Cellular elements of the blood could be seen passing rapidly through peritubular capillaries and individual glomerular capillary loops. With fluorescent filters in place, intravenously injected carboxyfluorescein was seen to pass through the glomerular capillary loops and then progressively through the different segments of the uriniferous tubules. Ligation of the renal artery resulted in rapid swelling of proximal tubule cells into the tubular lumens, loss of reflectiveness of the microvillous brush borders, and closure of the peritubular capillary spaces. Upon release of the ligature, the proximal tubule lumens again became patent, often opening up abruptly and in a zipper-like fashion down the length of the tubules. Increasing the glomerular filtration rate by intravenous infusion of mannitol resulted in increases in tubular luminal and perimeter dimensions. Mannitol also acted as an effective impermeant osmotic agent and prevented most of the cellular swelling which was otherwise seen in response to renal ischemia.

Animals

Continued contact lens wear in patients with giant papillary conjunctivitis.

We performed a six-year retrospective evaluation of 47 contact lens patients with a documented history of giant papillary conjunctivitis (GPC). The relationships between specific symptoms, contact lens history, treatment regimens, and clinical responses were studied. Specifically, we sought to determine the number of patients forced to discontinue contact lens wear due to GPC. Our study indicated two important preconditions to be associated with the onset of symptoms: the continued use of aging lenses and suboptimal lens care. For those patients treated and followed-up for at least 4 months, 79% obtained total resolution of their symptoms. Though the numbers are too small to make any significant conclusions about treatment methods it seems that lens discontinuance alone does not appear to be an adequate treatment for this condition and may serve better as adjunct to other therapy.

Adolescent