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Cornelia Corinne Drechsler, Alvena Kureshi, Stephan Reichl, Gerd Geerling, Julie Daniels, Stefan Schrader; Development of a conjunctival tissue substitute on the basis of plastic compressed collagen. Invest. Ophthalmol. Vis. Sci. 2013;54(15):2107.
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Ocular surface disorders can cause severe ocular surface inflammation, conjunctival scarring, fornix shortening and ankyloblepharon. There is the need for the development of new matrices for conjunctival reconstruction for this group of patients. Therefore, aim of this study was, to evaluate the suitability of compressed collagen as a substrate for the expansion of human conjunctival epithelial cells in order to develop an epithelialized conjunctival substitute for fornix reconstruction.
Human conjunctiva epithelial cells (HCEC) were cultured on acellular compressed collagen constructs or those containing human conjunctival fibroblasts (HCF). The epithelial cells were evaluated for cytokeratin expression (CK19 /CK4) and the appearance of goblet cells. The presence of putative progenitor epithelial cell markers such as p63α, ABCG2 and CK15 was assessed by immunostaining. The proliferative capacity of the epithelial cells after expansion on the plastic compressed collagen gels was evaluated by their colony forming efficiency and compared to cells cultured on amniotic membrane.
The results showed successful expansion of the conjunctival epithelial cells on both the acellular compressed collagen constructs and those containing human conjunctival fibroblasts. The immunofluorescence staining revealed the expression of the cytokeratins such as CK19 as well as the putative progenitor cell markers such as CK15 and ABCG2. After expansion on the collagen constructs, the conjunctival epithelial cell population was still able to form epithelial cell colonies, as tested by colony forming efficiency assay.
Plastic compressed collagen seems to be a suitable substrate for the expansion of human conjunctival epithelial cells. The presence of epithelial cells, positive for putative progenitor cell markers and the sustained proliferative capacity of the epithelial cell population indicates the maintenance of conjunctival cells with progenitor cell characteristics on the collagen gels. Therefore, an epithelialized conjunctival tissue construct on the basis of compressed collagen, might be a promising alternative bioartificial tissue substitute for conjunctival reconstruction.
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