A histochemical study of the distribution of dextran 500 in human corneas during organ culture

Redbrake, Claudia; Salla, Sabine; Nilius, Regina; Becker, Jakob; Reim, Martin

doi:10.1076/ceyr.16.5.405.7044

Cited by 32 publications

(28 citation statements)

References 8 publications

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“…An average of 8.4% EC loss was reported after 1–4 days of deswelling in an OC medium supplemented with 5% dextran T500 [7]. We previously reported a 15% cell loss after 2 days of deswelling in commercial media containing 5% dextran T500 [8,9], a result consistent with that of Rieger et al [10] and Redbrake et al [11] in similar conditions using homemade media containing 5% dextran T500. Consequently, daily EC loss in dextran-containing media appears much higher than average daily EC loss of 0.9% in dextran-free media used in the first phase of OC [8].…”

Section: Introductionsupporting

confidence: 79%

Poloxamines for Deswelling of Organ-Cultured Corneas

et al. 2012

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Background: Poloxamines are amphiphilic tetrofunctional block copolymers composed of four polyoxyethylene-polyoxypropylene arms joined to a central ethylene diamine bridge. Their safe profile allows diverse pharmaceutical and biomedical applications. Aim: To assess their use for corneal deswelling using a porcine model of organ culture (OC). Methods: Five poloxamines (T90R4, T904, T908, T1107 and T1307) were dissolved in a standard commercial OC medium (control) to reach 350 mosm kg^–1. In vitro cytotoxicity was tested using MTT assay on human corneal epithelial and endothelial cell (EC) lines and on primary human corneal fibroblasts. Paired porcine corneas stored in OC for 3 days were assigned for 48 h to a poloxamine medium or to a standard deswelling medium containing 5% dextran T500. Corneal EC density, morphometry, mortality, stromal thickness and transparency were evaluated before and after deswelling. Post-deswelling, EC viability/mortality was determined using a fluorescent live/dead assay. Results: Besides similar corneal thickness reduction and transparency improvement, T908, T1107 and T1307 decreased EC loss (5.4 ± 1.7% vs. 9.9 ± 2.6% in controls (p < 0.001)) and mortality, improved EC morphometry and reduced endothelial lesions compared to dextran. Conclusion: On this porcine model, poloxamines T908, T1107 and T1307 appear as good candidates to replace dextran for the deswelling. Experiments on human corneas are now necessary to confirm their efficiency and safety profile in OC.

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Section: Introductionsupporting

confidence: 79%

Poloxamines for Deswelling of Organ-Cultured Corneas

et al. 2012

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“…Other studies suggest a negative effect of dextran on the endothelial cell density, so this might be enhanced in case of exposure to a hypotonic sucrose solution [4,11,13]. However, the difference between the two groups was statistically not significant.…”

Section: Discussionmentioning

confidence: 91%

Endothelial cell density in porcine corneas after exposure to hypotonic solutions

Meltendorf

Ohrloff

Rieck

et al. 2006

Graefe's Arch Clin Exp Ophthalmol

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“…Furthermore, the influence of dextran accumulation, a high-molecular-weight polysaccharide, in the corneal tissue and dextran-induced intracellular edema of the keratocytes must be considered in the measured ablation patterns [25].…”

Section: Discussionmentioning

confidence: 99%

“…The corneas were stored in organ culture (modified Eagle's medium) at 32°C until use. Before the experiments the corneas were immersed in hyperosmolar solutions (6% dextran 500) for up to 48 h for deswelling to reach a corneal thickness of less than 800 mm [25,33]. Corneas with macroscopic visible corneal abnormalities were excluded from the experiment.…”

Section: Methodsmentioning

confidence: 99%