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Biomedical subjects

Claude J Giasson

Publications and source records attributed to Claude J Giasson.

5 recordsLinked to original sources

Autologous transplantation of rabbit limbal epithelia cultured on fibrin gels for ocular surface reconstruction.

PURPOSE: Regeneration of the corneal epithelium could be severely impaired in patients suffering from limbal stem cell deficiency. The purpose of this study was to evaluate the restoration of the corneal epithelium by grafting onto denuded corneas autologous limbal cells cultured on fibrin gels. The rabbit model was chosen to allow the microscopic evaluation over time after grafting. METHODS: Rabbit limbal epithelial cells (RLECs) were isolated and cultured from small limbal biopsies (3 mm2). The epithelium was separated from stroma after dispase digestion and put in culture on lethally irradiated fibroblasts used as a feeder layer. At the first passage, RLECs were cultured on a fibrin gel matrix. At confluence, the cultured epithelia were grafted in vivo on denuded autologous rabbit corneas. At different postoperative times, grafted and control (without graft or grafted with fibrin gels only) rabbit corneas were compared in vivo with a slit lamp microscope, and in situ by histological and immunohistological microscopy of harvested biopsies. RESULTS: A small limbal biopsy was sufficient to generate enough RLECs to prepare several grafts and to perform cell analysis. Only two weeks were required to produce a cultured epithelium suitable for autologous transplantation. One month after grafting, a normal corneal phenotype was observed on the ocular surface of grafted rabbits in contrast to the control rabbits (ungrafted or grafted with fibrin gel only) where histological signs of conjunctivalization were found. The absence of goblet cells and negative staining for keratin 4 confirmed that the cultured cells persisted and that the epithelium regenerated after grafting was not from conjunctival origin. CONCLUSIONS: Our results demonstrate that an autologous epithelium cultured on a physiologically biodegradable matrix can be prepared from a small biopsy and grafted on denuded cornea. The autologous graft allows epithelial regeneration from cultured cells and promotes corneal healing of unilateral total stem cell deficiency.

Animals↗

Morphometry of cells and guttae in subjects with normal or guttate endothelium with a contour detection algorithm.

PURPOSE: To develop a semiautomatic method to analyze morphology of cells and guttae in corneal endothelium. METHODS: Specular endothelial pictures from 42 and 21 subjects with healthy and guttate corneas, respectively, were analyzed independently by two observers with cell contour-extracting routines. One observer also analyzed healthy endothelia with the Corner method (Bambi). Differences between observers and between methods in mean cell area (MCA), coefficient of variation (CV), and percentage of cells with five, six, or seven sides were tested for significance with paired t tests. The Contour analysis of pictures with guttae included their mean area. RESULTS: There were no significant differences in MCA, CV, or the percentage of cells with five, six, or seven sides between the measurements obtained on repeated analysis by the same observer or on a second analysis performed by a different observer with the Contour method. However, the differences between the Contour and Bambi methods were statistically significant for MCA (337.5 +/- 37.7 vs. 327.7 +/- 36.5), CV (0.32 +/- 0.05 vs. 0.30 +/- 0.05), and percentage of cells with six and seven sides, but not for the percentage of five-sided cells. In subjects with guttata, the MCA was 561 +/- 170 microm, and the mean area of guttae was 1,538 +/- 849 microm. CONCLUSIONS: This detection algorithm is repeatable and reproducible, and it generates a cell border overlay useful in analyzing the morphology of cells and guttae. The analysis of corneal guttae could become a useful follow-up procedure to discriminate between patients with corneal guttata and Fuchs dystrophy.

Adult↗

Overwear of contact lenses: increased severity of clinical signs as a function of protein adsorption.

This study compared the clinical behavior of disposable and frequent replacement Acuvue and 1-Day contact lenses. Each type of lens was worn on one eye according to the schedule recommended by the manufacturer, and on the other eye for a longer period of time, up to 30 days in length. Both type of lenses were prescribed on a daily-wear basis. The amount of protein collected from the lenses was measured using two spectrophotometric protein assays. Visual acuity and comfort, along with several other clinical signs, were classified according to Cornea and Contact Lens Research Unit (CCLRU) scales, and possible associations between each of these signs and the amount of protein extracted from the lenses was tested. A comparison between the lens worn on the compliant eye with the lens worn on the noncompliant eye allowed us to measure the impact of overwear on ocular health and subjective clinical findings. After four months of study, the overwear of Acuvue and 1-Day lenses significantly increased the amount of protein bound on the contact lenses, as well as the severity of upper conjunctival papillae, upper lid conjunctival hyperemia, and limbal congestion. Even if reduced values for visual acuity and noninvasive break-up time (NIBUT) were identified, these variations were not found to be statistically significant. The clinical implications of this study would allow a practitioner to identify, according to the variations in several clinical signs, a patient who overwears contact lenses, so that action may be taken to reduce possible deleterious effects on ocular health.

Adsorption↗

[Amnion and ocular surface problems].

The amniotic membrane, the most internal placental membrane, has various properties useful in ophthalmology. Collected on delivery by elective Caesarean section, the amnion is prepared under sterile conditions, and, usually, cryopreserved until its use as a biological bandage or as a substrate for epithelial growth in the management of various ocular surface conditions. Specifically, the amnion is used to : (1) limit formation of adhesive bands between eyelids and eyeball (symblepharon) or the progression of a fibrovascular outgrowth towards the cornea (pterygium) or to (2) facilitate the healing of corneal ulcers, bullous keratopathy, and corneal stem cell deficiency. In this last condition, either hereditary or acquired after a thermal or a chemical burn, corneal stem cells, located at a transitional zone between the cornea and conjunctiva, are lost. These cells are essential for renewal of corneal epithelium in normal and in diseased states. The loss of these cells leaves the corneal surface free for invasion by conjunctival epithelium. Not only, does conjunctival epithelium support the development of vascularisation on the normally avascular cornea, but some conjunctival cells differentiate into mucus secreting goblet cells. Such a change in phenotype leads to loss of corneal transparency and visual disability. The removal of this fibro-vascular outgrowth in combination with transplantation of both amniotic membrane and corneal stem cells are used to treat this condition. The amnion stimulates the proliferation of less differentiated cells which have the potential to reconstruct the cornea. This potential is at the origin of the hypothesis that the amnion may provide an alternative niche for limbal stem cells of the corneal epithelium. It abounds in cytokines and has antalgic, anti-bacterial, anti-inflammatory and anti-immunogenic properties, in addition to allowing, like fetal skin does, wound healing with minimal scar formation. These desirable properties are responsible for the increasing use of amniotic membrane in ophthalmology. The complete understanding of the mechanisms of action of amniotic membrane for ocular surface diseases has yet to be understood. Once revealed by research, they may provide new pharmacological avenues to treat ocular surface diseases.

Amnion↗