Ewa Wroblewska-Czajka

Cultivated Oral Mucosa Epithelium in Ocular Surface Reconstruction in Aniridia Patients

Purpose. Efficacy of cultivated oral mucosa epithelial transplantation (COMET) procedure in corneal epithelium restoration of aniridia patients. Methods. Study subjects were aniridia patients (13 patients; 17 eyes) with irregular, vascular conjunctival pannus involving visual axis who underwent autologous transplantation of cultivated epithelium. For the procedure oral mucosa epithelial cells were obtained from buccal mucosa with further enzymatic treatment. Suspension of single cells was seeded on previously prepared denuded amniotic membrane. Cultures were carried on culture dishes inserts in the presence of the inactivated with Mitomycin C monolayer of 3T3 fibroblasts. Cultures were carried for seven days. Stratified oral mucosa epithelium with its amniotic membrane carrier was transplanted on the surgically denuded corneal surface of aniridia patients with total or subtotal limbal stem cell deficiency. Outcome Measures. Corneal surface, epithelial regularity, and visual acuity improvement were evaluated. Results. At the end of the observation period, 76.4% of the eyes had regular transparent epithelium and 23.5% had developed epithelial defects or central corneal haze; in 88.2% of cases visual acuity had increased. VA range was from HM 0.05 before the surgery to HM up to 0.1 after surgery. Conclusion. Application of cultivated oral mucosa epithelium restores regular epithelium on the corneal surface with moderate improvement in quality of vision.

Clinical, confocal, and morphological investigations on the cornea in human mucopolysaccharidosis IH-S.

Purpose: The aim was to describe the confocal and histological findings of 2 corneas from a patient with an advanced case of type I mucopolysaccharidoses Hurler–Scheie disease (MPS IH-S). Methods: Both corneas from an MPS IH-S–affected patient were examined in vivo using confocal microscopy and then removed and processed for evaluation using light microscopy and transmission and scanning electron microscopy. Results: Confocal microscopy evaluation showed basal epithelial cells with either diffuse or granular hyperreflectivity. Keratocytes were highly reflective determining a web-shaped stromal appearance. Endothelial cells were barely visible. The histopathological study demonstrated superficial cells with apical microfolds, small vesicles, and evident intercellular junctions. The wing cells showed either well-evident tonofilaments and small peripheral vesicles, or large paranuclear vesicles. The basal cells showed polygonal shapes, many small vesicles, and enlarged intercellular spaces. The Bowman layer was either normal or thinner and was formed by variably electron dense material. In the stroma, irregularly oriented lamellae, many vesicle-filled keratocytes, and intercellular granular material were present. The Descemet membrane was normal, whereas the corneal endothelium showed marked degenerative changes. Conclusions: The confocal alterations appeared consequent to the anomalous accumulation of material. The histopathological images gave a clue to better understand the corneal changes demonstrated by the confocal studies in MPS IH-S.

Donor disc attachment assessment with intraoperative spectral optical coherence tomography during descemet stripping automated endothelial keratoplasty.

Optical coherence tomography has already been proven to be useful for pre- and post-surgical anterior eye segment assessment, especially in lamellar keratoplasty procedures. There is no evidence for intraoperative usefulness of optical coherence tomography (OCT). We present a case report of the intraoperative donor disc attachment assessment with spectral-domain optical coherence tomography in case of Descemet stripping automated endothelial keratoplasty (DSAEK) surgery combined with corneal incisions. The effectiveness of the performed corneal stab incisions was visualized directly by OCT scan analysis. OCT assisted DSAEK allows the assessment of the accuracy of the Descemet stripping and donor disc attachment.

Phenotype and genotype analysis in patients with macular corneal dystrophy

Aim The aim of this study was to analyse corneal morphological organisation and identify mutations in the carbohydrate sulfotransferase 6 gene (CHST6) in patients with macular corneal dystrophy originating in a Polish population. Methods Macular corneal dystrophy was diagnosed in 24 patients based on the slit-lamp exam, confocal microscopy, 1310 nm time domain and 840 nm spectral domain optical coherence tomography. 10 corneal buttons obtained from penetrating keratoplasty were processed for light microscopy. Genetic analysis of the CHST6 gene was performed, followed by a study of the sequencing results. Results Highly reflective, diffuse corneal deposits and a general increase in reflectivity were revealed with optical coherence tomography and confocal microscopy. The deposits extended from the Bowman layer to the Descemet membrane and correlated with the Alcian blue-positive granular-filamentous material into and around the stromal keratocytes confirmed by structural analysis of the corneal buttons. The genetic analysis of the blood samples identified the following mutations and single nucleotide polymorphisms: novel P64L (heterozygous), Y110C (homozygous), R162G and L200R, and M1L (heterozygous and homozygous). Conclusions Genetic mutation heterogeneity was revealed. No phenotype heterogeneity was revealed among patients with in vivo corneal morphology assessment or histological analysis.

Total Penetrating Keratoplasty: Indications, Therapeutic Approach, and Long-Term Follow-Up

Purpose Evaluation of the indications, anatomical and functional results, and complications of total penetrating keratoplasty (TPK) in disorders involving whole cornea. Materials and Methods We analyzed outcomes of the surgical treatment of 47 eyes of 46 patients that underwent TPK. Indications were infectious keratitis, autoimmune disease, injury of the eyeball (mainly chemical burns), and other combined disorders. The surgical technique involved dissection of affected tissues with a margin of 1.0 mm. The size of the corneal graft ranged from 10.0 to 14.0 mm. We analyzed indications, outcomes, and complications of surgery. Results Final restoration of the ocular integrity and maintenance of the globe were achieved in 27 eyes (57%). More than one surgery was necessary in a total of 29 eyes (62%). The frequency of retransplantations did not vary significantly between the groups with different causes of corneal melting/perforation (63% of eyes with infection, 66% of eyes after trauma and 70% of eyes of patients with autoimmune disorders). Surgical treatment failed in 20 eyes (43%). Evisceration was necessary in 13 eyes (28%), phthisis occurred in 7 cases (15%). Conclusion TPK should be considered as a last line treatment in huge corneal destruction to restore integrity of the eye globe.

Comparison of the Techniques of Secondary Intraocular Lens Implantation after Penetrating Keratoplasty

Aim To conduct a retrospective analysis of secondary IOL implantation in patients who underwent PK with no simultaneous IOL implantation. Materials and Methods The retrospective study of the secondary implantation of IOLs was conducted in 46 eyes that underwent a primary operation with PK and cataract/lens extraction with no IOL implantation due to capsule rupture or combining corneal or intraocular complications. The minimum period from PK was 12 months. All secondary IOL implantations were performed from January 2011 to August 2017. Aphakic postkeratoplasty patients were treated using one of the surgical techniques for secondary IOL implantation. In-the-bag IOL implantation was possible if the posterior capsule was complete. If the lens capsule remnants were sufficient to provide secure IOL support, an in-the-sulcus IOL implantation was performed. Scleral fixation was offered in eyes with extensive capsular deficiency or the presence of the vitreous body in anterior chamber. BCVA and expected and achieved refraction were evaluated; we included using two biometry devices, and results were compared. Results The corrected distance visual acuity (CDVA) before surgery ranged from 0.1 to 0.8 (mean 0.54 ± 0.17). After secondary IOL implantation, CDVA ranged from 0.2 to 0.8 (mean 0.43 ± 0.14) at postoperative 1 month and from 0.3 to 0.9 (mean 0.55 ± 0.15) at postoperative 6 months (p < 0.05). Comparison of the final refraction using two methods of biometry showed no statistically significant difference in the group that underwent scleral fixation of the IOL, similar to the findings for the in-the-bag and in-the-sulcus IOL implantation groups. In the scleral-fixation group, p=0.55 for the USG biometry technique and p=0.22 for the OB technique. p values for the IOL-implantation group were p=0.49 and p=0.44, respectively. Conclusion Both implantation methods are safe for the patients. Final refraction is depending on the technique and indication to keratoplasty. Both biometry techniques deliver precise data for IOL choice.

Cytoarchitecture of epithelial inflammatory infiltration indicates the aetiology of infectious keratitis

To analyse cytological features of corneal epithelium in infectious keratitis.