Spontaneous full thickness macular hole development and closure in a patient with nucleus dislocation due to hypermature cataract: a case report.

Spontaneous posterior capsule rupture with lens-nucleus dislocation is a very rare entity, as is the development and spontaneous closure of a full thickness macular hole (FTMH) after vitrectomy. The occurrence of these two entities in one eye has not been previously described. A 79-year-old woman was referred because of the right eye intermittent pain and progressive visual loss. Best corrected visual acuity (BCVA) with correction for aphakia was 20/20. Intraocular pressure was normal with therapy. The cornea, anterior chamber, and vitreous were clear. Gonioscopy was normal. The capsular bag was clear, with rolled-up anterior and posterior lens capsule, and the nucleus dislocated in the vitreous. As surgery waiting time was prolonged due to administrative problems, the patient's intraocular pressure (IOP) increased and cystoid macular edema (CME) with lamellar macular hole developed. The patient underwent pars plana vitrectomy with endophacofragmentation and epiretinal membrane peeling. Postoperative optical coherence tomography was normal, BCVA was 20/40, and IOP was normal with topical therapy. One month after surgery, the eye was without signs of inflammation and IOP started rising in spite of maximum therapy. CME reoccurred and progressed to a FTMH, which started closing spontaneously in one month. One year after surgery, IOP normalized and FTMH closed completely. A dislocated crystalline lens in a quiet eye with normal BCVA, which rapidly developed into intractable glaucoma and FTMH, is an unusual finding. The deterioration was followed by spontaneous IOP normalization and macular hole closure. Such unexpected disease course, suggesting a possible autoimmune reaction, has not yet been described.

Identification and characterization of dendritic cell subtypes in preserved and cultured cadaveric human corneolimbal tissue on amniotic membrane.

PURPOSE: Rejection is the leading cause of failure of limbal allogafts. Resident dendritic cell (DC) maturation plays a critical role in host allosensitization. There are two lineages: myeloid (mDC) and lymphoid (pDC), with different biological properties. The aim was to analyse the distribution of DC subtypes in limbal explant cultures on amniotic membrane (AM), cultivated on either the epithelial or stromal side and to compare the results with directly isolated cells from cadaveric whole corneoscleral tissue divided into specific areas. METHODS: The expression of CD11c (mDC), CD303/CD123 (pDC) and costimulatory molecules CD80, CD86 and activation markers HLA-DR, CD83 was investigated by flow cytometry. Additionally, the corneal epithelium marker CK12 and ABCB5, a new epithelial stem cell marker, were investigated. RESULTS: Cells positive for pDC and mDC markers were found in all examined areas, with a nonsignificant prevalence of pDC. In limbal explant cultures on AM, the percentage of pDC and mDC was similar, with no statistically significant difference between cultures on epithelial or stromal sides of AM. However, with ex vivo limbal explant cultivation on AM, the pDC content declined significantly (p < 0.05) and the ABCB5 marker was likewise statistically significantly reduced. CONCLUSION: This is the first study to characterize the distribution of pDC and mDC subsets in cultured and noncultured human corneolimbal tissue. Additionally, ABCB5 positive cells were identified. These findings might be important for future strategies, allowing preparation of corneolimbal allografts with optimal stem cell content for a longer lasting therapeutic effect.

Preservation of Ocular Epithelial Limbal Stem Cells: The New Frontier in Regenerative Medicine.

Significant advances have been made in the field of ocular regenerative medicine. Promising stem cell-based therapeutic strategies have been translated into the clinical practice over the last few decades. These new stem cell-based therapies offer the possibility of permanently restoring corneal epithelium in patients with severe disabling and blinding ocular surface disease. The European Union has already classified stem cell-based therapies as "medicinal products". Therefore, manipulation is strictly regulated according to the defined conditions of good manufacturing practice, with the production of stem cell therapeutics at only accredited production sites authorized by the national regulatory agencies. In this regard, as first medical products are licensed for commercial use in Europe enabling a more widespread access to a stem cell-based therapy, the need for safe, validated and reproducible techniques for ex vivo cultured tissue preservation and distribution are coming to the forefront of research. However, these provide various new challenges for biobanking industry such as the retention of viability, good functionality of stem cells and sterility issues. This chapter provides an overview of the current advances in the field of corneal/limbal epithelial stem cell culture preservation techniques using either hypothermic storage or cryopreservation methods, that were used in different culturing steps (from stem cell isolation to the ex vivo epithelial graft preparation), with the reported impact on the post-thawing product recovery.

Effect of Cryopreserved Amniotic Membrane Orientation on the Expression of Limbal Mesenchymal and Epithelial Stem Cell Markers in Prolonged Limbal Explant Cultures.

PURPOSE: To evaluate the effect of prolonged limbal explants cultured without any scaffolds or on amniotic membrane (AM) on the viability, proliferation and differentiation potential of putative phenotypically defined cultured limbal mesenchymal (LMSC) and epithelial stem cells (LESC). METHODS: Limbal explants were cultivated on cryopreserved intact AM or plastic plates using medium supplemented with only human serum. AM was positioned with either the epithelial or stromal side up. The outgrowing cells were immunophenotyped for the co-expression of mesenchymal stem cell markers (CD73/CD90/CD105 positive and CD45 negative), proliferation and putative progenitor markers (CXCR4, CD117), epithelial markers and antigen presenting cell markers (CD80, CD83, CD86) by flow cytometry. Immunohistochemistry on limbal cultures cultivated on AM was carried out with antibodies against pan-cytokeratin, p63, Ki67. RESULTS: Morphological and immunostaining analyses revealed two distinct stem cell population types, which could be identified over prolonged culturing time periods. Expression of LMSC markers and CXCR4 was significantly higher (p < 0.05) in cultures cultivated without AM. However, no statistically significant difference was observed in CD117 expression. The cells cultivated on AM retained an epithelial cell structure, which was further confirmed by histology examination. Histology revealed limbal epithelial growth and p63, Ki67 positive cells on both sides of AM. CONCLUSION: Limbal cells cultivated on AM exhibited a lower expression profile of LMSC and CXCR4 markers as limbal cells cultivated on plastic culture plates. However, CD117 expression was similar. Histology confirmed limbal epithelial cell growth on both sides of AM, with no morphological differences, or positivity of cells for p63 and Ki67.

Amniotic membrane properties and current practice of amniotic membrane use in ophthalmology in Slovenia.

Amniotic membrane (AM) is the innermost, multilayered part of the placenta. When harvested, processed and stored properly, its properties, stemming from AM biological composition, make it a useful tissue for ophthalmic surgery. AM was shown to have several beneficial effects: it promotes epithelization, has antimicrobial effects, decreases inflammation, fibrosis and neovascularization. Many case reports and case series as well as practical experience (e.g. reconstruction of conjunctival and corneal defects, treatment of corneal ulcers) demonstrated the beneficial effect of AM for different ophthalmological indications. The combination of the above mentioned beneficial effects and reasonable mechanical properties are also the reason why AM is used as a substrate for ex vivo expansion of epithelial progenitor cells. Recently, amnion-derived cells, which also have stem cell characteristics, have been proposed as potential contributors to cell-based treatment of ocular surface disease. However, the use of AM remains one of the least standardized methods in ophthalmic surgery. In this review, the various properties of AM and its current clinical use in ophthalmology in Slovenia are discussed.

Lid-parallel conjunctival folds (LIPCOF) and dry eye: a multicentre study.

AIMS: The study was designed to test the clinical application of the grading of lid-parallel conjunctival folds (LIPCOF) as a diagnostic test for dry eye. METHODS: At 12 centres in 11 countries, 272 eyes of 272 dry eye patients (75 men, 197 women) were examined. Their mean age was 52.7±16.2 years. The LIPCOF were graded according to the method of Höh et al. The tear film break-up time (BUT) was measured, and fluorescein staining and the Schirmer 1 test were performed. The subjective symptoms were evaluated by 16 questions. RESULTS: The LIPCOF score demonstrated significant positive correlations with age, dry eye disease severity and fluorescein staining (r>0.2, p<0.001), and negative correlations with BUT and results of the Schirmer 1 test (r<-0.2, p<0.001). The LIPCOF score exhibited a significant correlation with the overall subjective symptoms (r=0.250, p<0.001). The sensitivity and specificity of LIPCOF grading for discriminating between normal and dry eyes were best with the cut-off between LIPCOF degrees 1 and 2. CONCLUSIONS: The displayed medium sensitivity and specificity, and good positive predictive value of the LIPCOF test support the use of LIPCOF grading as a simple, quick and non-invasive dry eye screening tool.

Effect of soft contact lenses on the measurements of intraocular pressure with non-contact pneumotonometry.

PURPOSE: To evaluate the effect of soft contact lenses with different lens power on the measured value of the intraocular pressure with non-contact pneumotonometry. METHODS: 120 eyes (80 healthy volunteers: 50 women, 30 men, aged 22 to 61 years) were included in this study. Intraocular pressure was measured with a pneumotonometer Canon X-10 before and after insertion of the Ciba Vision Focus Night&Day soft contact lens. We used contact lenses with different lens power of + 0.25 D, + 1.00 D, + 4.00 D, - 1.00 D, and - 4.00 D. The averages of three measurements were taken as representative IOP values that were statistically evaluated. RESULTS: IOP measured over myopic lenses of - 1.00 and - 4.00 D showed lower values within the mean range of 1 mm Hg. The difference between the measurements over the myopic lenses was mostly smaller than +/- 2 mm Hg (78% when using - 1.00 D and 90% when using - 4.00 D contact lens). All the differences using + 0.25 D contact lens were smaller than +/- 2 mm Hg. The difference was considerably higher in measurements over + 1.00 and + 4.00 hyperopic contact lenses and showed strong increase with the lens power. CONCLUSIONS: In our study we showed that intraocular pressure can be reliably measured with non-contact pneumotonometry over myopic lenses or hypermetropic lenses with small lens power. This suggests that non-contact pneumotonometry is a useful method in patients wearing therapeutic contact lenses and contact lens wearers who, when measuring the eye pressure, would not need to remove the contact lenses before the examination.