Influence of Organ Culture on the Characteristics of the Human Limbal Stem Cell Niche.

Organ culture storage techniques for corneoscleral limbal (CSL) tissue have improved the quality of corneas for transplantation and allow for longer storage times. Cultured limbal tissue has been used for stem cell transplantation to treat limbal stem cell deficiency (LSCD) as well as for research purposes to assess homeostasis mechanisms in the limbal stem cell niche. However, the effects of organ culture storage conditions on the quality of limbal niche components are less well described. Therefore, in this study, the morphological and immunohistochemical characteristics of organ-cultured limbal tissue are investigated and compared to fresh limbal tissues by means of light and electron microscopy. Organ-cultured limbal tissues showed signs of deterioration, such as edema, less pronounced basement membranes, and loss of the most superficial layers of the epithelium. In comparison to the fresh limbal epithelium, organ-cultured limbal epithelium showed signs of ongoing proliferative activity (more Ki-67+ cells) and exhibited an altered limbal epithelial phenotype with a loss of N-cadherin and desmoglein expression as well as a lack of precise staining patterns for cytokeratin ((CK)14, CK17/19, CK15). The analyzed extracellular matrix composition was mainly intact (collagen IV, fibronectin, laminin chains) except for Tenascin-C, whose expression was increased in organ-cultured limbal tissue. Nonetheless, the expression patterns of cell-matrix adhesion proteins varied in organ-cultured limbal tissue compared to fresh limbal tissue. A decrease in the number of melanocytes (Melan-A+ cells) and Langerhans cells (HLA-DR+, CD1a+, CD18+) was observed in the organ-cultured limbal tissue. The organ culture-induced alterations of the limbal epithelial stem cell niche might hamper its use in the treatment of LSCD as well as in research studies. In contrast, reduced numbers of donor-derived Langerhans cells seem associated with better clinical outcomes. However, there is a need to consider the preferential use of fresh CSL for limbal transplants and to look at ways of improving the limbal stem cell properties of stored CSL tissue.

[Conjunctival tumors in children : Histopathological diagnoses in 262 cases]. / Bindehauttumoren bei Kindern : Histopathologische Diagnosen in 262 Fällen.

BACKGROUND: The spectrum of conjunctival tumors in children and adolescents significantly differs from that of adults. OBJECTIVE: The aim of this study was to show the distribution of histopathological diagnoses of pediatric conjunctival tumors. MATERIAL AND METHODS: Retrospective case series including all conjunctival tumors of children and adolescents under 18 years of age examined histologically over a period of 25 years (1990-2015) at the Eye Center in Freiburg, Germany. RESULTS: Among the 262 conjunctival specimens, the most common diagnoses were conjunctival nevi (148, [56%]), most often with junctional activity (106/148, [72%]), pyogenic granulomas (26, [10%]), and dermolipomas (18, [7%]). In one case a malignant tumor (conjunctival melanoma) was detected (1, < 1%). CONCLUSION: The results suggest that pediatric conjunctival tumors are benign in most cases. Most of them were of melanocytic origin, with a junctional conjunctival nevus being the most common diagnosis.

Histological Diagnosis of Ocular and Periocular Tuberculosis 1945 - 2020. / Histologisch diagnostizierte okuläre und periokuläre Tuberkulose von 1945 bis 2020.

BACKGROUND: Ocular tuberculosis is a rare but important differential diagnosis for inflammatory conditions of all eye tissues, including the ocular surface and adnexa. Tissue diagnostics may prove challenging as some ocular tissues are difficult to biopsy and the detection of pathogens may be insensitive. We were interested in how many cases in the archive of the ophthalmopathological laboratory had been diagnosed with (peri)ocular tuberculosis since 1945. MATERIALS AND METHODS: Retrospective analysis of historical records and specimens of the ophthalmopathology laboratory of the eye department at Freiburg university hospital. Systematic re-evaluation of available slides for presence of granuloma, necrosis, giant cells, acid fast bacteria, and chronic as well as acute inflammation, plus comparison of current and historic evaluations. In addition, we describe a recent case with tuberculoma of the iris. RESULTS: There were 50,418 records archived since 1945, of which 23 specimens taken from 22 patients had been diagnosed as (peri)ocular tuberculosis. Of these, 22 (96%) were archived and available for re-interpretation. Four specimens (17%) had been excised from children. The most common tissues were enucleated eye globes (10/23, 44%), followed by the lacrimal sac (5/23, 22%) and conjunctiva (2/23, 9%). The most frequent histopathological findings were granulomas (23/23, 100%), chronic inflammation (22/23, 96%), giant cells (21/23, 91%), and necrosis (14/23, 61%). An acute inflammatory response was found in 4/23 specimens (17%). Ziehl-Neelsen stains for acid-fast bacteria had been performed in five cases, of which three were positive (60%). The greatest discrepancy between current and historical findings related to the presence of necrosis (59% consensus). In other findings, the consensus was high (78 - 96%). In a recent case of a patient with wasting syndrome attributed to lymphoma, histopathological workup of an iris tumour led to the diagnosis of tuberculosis. CONCLUSION: Ocular tuberculosis is a rare but important histopathological differential diagnosis. In the available specimens, the classic finding of necrotizing inflammation was rarest and showed least consensus on histological re-evaluation. Other typical findings, such as giant cells and a predominantly lymphocytic infiltrate, are sometimes not found even with proven presence of Mycobacterium tuberculosis. They should not be considered essential in cases where there is strong clinical suspicion.